LOCUS       pPC1540                10257 bp ds-DNA     circular SYN 05-AUG-2021
DEFINITION  [pBR322, Cb] pCMV-bpNLS-SpCas9(R221K N394K
            H840A)-SGGSx2-sv40NLS-SGGSx2-MMLV_RT_pentamutant_GScodon-bpNLS(fixe
            d)-cmycNLS-P2A-BSD (PEmax-P2A-BSD).
ACCESSION
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct.
REFERENCE   1  (bases 1 to 9753)
  AUTHORS   AVA
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Mar 20, 2019 from SnapGene 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     enhancer        1..380
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        381..584
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /label="CMV promoter"
     primer_bind     468..492
                     /Sequence="GGGAGTTTGTTTTGGCACCAAAATC"
                     /%GC="44.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ676"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1g-9qpay3y"">KZ676</a>"
                     /label="KZ676"
     primer_bind     535..555
                     /%GC="66.7"
                     /Hairpin_Tm="52.4"
                     /Self_Dimer_Tm="3.5"
                     /Tm="67.0"
                     /Sequence="CGCAAATGGGCGGTAGGCGTG"
                     /created_by="primer3"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="BP0002 (CMV-For)"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:e-9439785"">BP0002 (CMV-For)</a>"
                     /label="BP0002 (CMV-For)"
     primer_bind     552..583
                     /Sequence="CGTGTACGGTGGGAGGTCTATATAAGCAGAGC"
                     /Tm="68.8"
                     /%GC="53.1"
                     /Hairpin_Tm="36.4"
                     /Self_Dimer_Tm="4.3"
                     /created_by="chenp"
                     /label="PC1666"
     primer_bind     complement(552..583)
                     /Sequence="GCTCTGCTTATATAGACCTCCCACCGTACACG"
                     /Tm="68.8"
                     /%GC="53.1"
                     /Hairpin_Tm="39.4"
                     /Self_Dimer_Tm="None"
                     /created_by="chenp"
                     /label="PC1667"
     primer_bind     complement(559..587)
                     /Sequence="ACCAGCTCTGCUTATATAGACCTCCCACC"
                     /%GC="51.7"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0767"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:198-a5u3wkd"">PC0767</a>"
                     /label="PC0767"
     primer_bind     576..602
                     /Sequence="AGCAGAGCTGGUTTAGTGAACCGTCAG"
                     /%GC="51.9"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0766"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:193-a5u3wk9"">PC0766</a>"
                     /label="PC0766"
     promoter        626..644
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     626..645
                     /Sequence="TAATACGACTCACTATAGGG"
                     /%GC="40.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="BP0012 (T7 Promoter)"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2p-93oy69x"">BP0012 (T7 Promoter)</a>"
                     /label="BP0012 (T7 Promoter)"
     primer_bind     626..645
                     /Sequence="TAATACGACTCACTATAGGG"
                     /%GC="40.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1497"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1d-cej84u7"">PC1497</a>"
                     /label="PC1497"
     primer_bind     complement(634..659)
                     /Sequence="ATGGTGGCGGCUCTCCCTATAGTGAG"
                     /%GC="57.7"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0626"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:88-9pors9o"">PC0626</a>"
                     /label="PC0626"
     primer_bind     complement(636..659)
                     /Sequence="ATGGTGGCGGCUCTCCCTATAGTG"
                     /%GC="58.3"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1059"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:5v-bljrmru"">PC1059</a>"
                     /label="PC1059"
     primer_bind     complement(643..659)
                     /Sequence="ATGGTGGCGGCUCTCCC"
                     /%GC="70.6"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0190"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2u-96urery"">PC0190</a>"
                     /label="PC0190"
     primer_bind     complement(645..691)
                     /Sequence="ACTCGAACTCGCTUCCGTCGGCTGTCCGTTTCATGGTGGCGGCTCTC
                     "
                     /%GC="61.7"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0234"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1x2-998axgr"">PC0234</a>"
                     /label="PC0234"
     primer_bind     648..676
                     /Sequence="AGCCGCCACCAUGAAACGGACAGCCGACG"
                     /%GC="65.5"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0198"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:327-97kkp7h"">PC0198</a>"
                     /label="PC0198"
     CDS             658..714
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="MKRTADGSEFESPKKKRKV"
                     /label="SV40 NLS"
     primer_bind     674..706
                     /Sequence="ACGGAAGCGAGUTCGAGTCACCAAAGAAGAAGC"
                     /%GC="51.5"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0738"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1ci-a43x1p9"">PC0738</a>"
                     /label="PC0738"
     primer_bind     complement(684..713)
                     /Sequence="ACTTTCCGCTUCTTCTTTGGTGACTCGAAC"
                     /%GC="46.7"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0831"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1i4-acth48u"">PC0831</a>"
                     /label="PC0831"
     CDS             715..4815
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from
                     theStreptococcus pyogenes Type II CRISPR/Cas system"
                     /note="generates RNA-guided single strand nicks in DNA
                     dueto the H840A mutation in the HNH catalytic domain"
                     /Original_Translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLG
                     NTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD
                     DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLR
                     LIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKA
                     ILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSK
                     DTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYD
                     EHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD
                     GTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEK
                     ILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDK
                     NLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRK
                     VTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDIL
                     EDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ
                     SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPA
                     IKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKE
                     LGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLK
                     DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERG
                     GLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVS
                     DFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMI
                     AKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDF
                     ATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT
                     VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLII
                     KLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ
                     KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL
                     FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
                     /modified_by="chenp"
                     /label="Cas9(R221K N394K H840A)"
     primer_bind     715..735
                     /Sequence="GACAAGAAGTACAGCATCGGC"
                     /%GC="52.4"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1498"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1i-cej84ud"">PC1498</a>"
                     /label="PC1498"
     primer_bind     complement(722..743)
                     /Sequence="ATGTCCAGGCCGAUGCTGTACT"
                     /%GC="54.5"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0292"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1o-9atlcx0"">PC0292</a>"
                     /label="PC0292"
     primer_bind     725..749
                     /Sequence="ACAGCATCGGCCUGGACATCGGCAC"
                     /%GC="64.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1335"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:93-cczyqbw"">PC1335</a>"
                     /label="PC1335"
     primer_bind     730..747
                     /Sequence="ATCGGCCTGGACAUCGGC"
                     /%GC="66.7"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0293"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1t-9atlcx8"">PC0293</a>"
                     /label="PC0293"
     primer_bind     750..774
                     /Sequence="CAACTCTGTGGGCTGGGCCGTGATC"
                     /%GC="64.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ678"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1q-9qpay45"">KZ678</a>"
                     /label="KZ678"
     primer_bind     complement(793..814)
                     /Sequence="GCACCTTGAATTTCTTGCTGGG"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1499"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1n-cej84uf"">PC1499</a>"
                     /label="PC1499"
     primer_bind     complement(854..878)
                     /Sequence="CCGCTGTCGAACAGCAGGGCTCCGA"
                     /%GC="68.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ677"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1l-9qpay42"">KZ677</a>"
                     /label="KZ677"
     primer_bind     complement(1096..1115)
                     /Sequence="ATGGTGGGGTACUTCTCGTG"
                     /%GC="55.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0229"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1wd-998axgd"">PC0229</a>"
                     /label="PC0229"
     primer_bind     1103..1122
                     /Sequence="AGTACCCCACCAUCTACCAC"
                     /%GC="55.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0228"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1w8-998axgb"">PC0228</a>"
                     /label="PC0228"
     primer_bind     1132..1151
                     /Sequence="AAACTGGTGGACAGCACCGA"
                     /Hairpin_Tm="51.8"
                     /Tm="62.0"
                     /Self_Dimer_Tm="None"
                     /%GC="55.0"
                     /created_by="primer3"
                     /label="PC0872"
     primer_bind     1132..1151
                     /Sequence="AAACTGGTGGACAGCACCGA"
                     /%GC="55.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0872"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:89-b14udte"">PC0872</a>"
                     /label="PC0872"
     primer_bind     complement(1195..1214)
                     /Sequence="AAGTGGCCCCGGAACTTGAT"
                     /Hairpin_Tm="33.0"
                     /Tm="61.8"
                     /Self_Dimer_Tm="0.7"
                     /%GC="55.0"
                     /created_by="primer3"
                     /label="PC0873"
     primer_bind     complement(1195..1214)
                     /Sequence="AAGTGGCCCCGGAACTTGAT"
                     /%GC="55.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0873"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8e-b14udti"">PC0873</a>"
                     /label="PC0873"
     primer_bind     1249..1269
                     /Sequence="GACGTGGACAAGCTGTTCATC"
                     /%GC="52.4"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1500"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1s-cej84uh"">PC1500</a>"
                     /label="PC1500"
     primer_bind     1250..1274
                     /Sequence="ACGTGGACAAGCTGTTCATCCAGCT"
                     /%GC="52.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ679"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1v-9qpay4b"">KZ679</a>"
                     /label="KZ679"
     primer_bind     complement(1345..1376)
                     /Sequence="AGCTTTCTGCTCTTGCTCAGTCTGGCAGACAG"
                     /Tm="71.1"
                     /%GC="53.1"
                     /Hairpin_Tm="52.7"
                     /Self_Dimer_Tm="40.1"
                     /created_by="chenp"
                     /label="PC1651"
     USER            1364..1376
                     /created_by="chenp"
     primer_bind     1364..1396
                     /Sequence="AGAGCAGAAAGCTGGAAAATCTGATCGCCCAGC"
                     /Tm="71.5"
                     /%GC="51.5"
                     /Hairpin_Tm="58.2"
                     /Self_Dimer_Tm="29.7"
                     /created_by="chenp"
                     /label="PC1650"
     misc            1372..1374
                     /created_by="chenp"
                     /modified_by="chenp"
                     /label="R221K"
     misc_feature    1372..1373
                     /Original_Bases="CG"
                     /label="CG"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     1750..1774
                     /Sequence="TACAAAGAGATTTTCTTCGACCAGA"
                     /%GC="36.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ680"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:20-9qpay4h"">KZ680</a>"
                     /label="KZ680"
     primer_bind     complement(1872..1904)
                     /Sequence="AGGTCCTCTCTCTTCAGCTTCACGAGCAGTTCC"
                     /Tm="71.4"
                     /%GC="54.5"
                     /Hairpin_Tm="54.3"
                     /Self_Dimer_Tm="0.5"
                     /created_by="chenp"
                     /label="PC1649"
     primer_bind     1879..1908
                     /Sequence="CTCGTGAAGCTGAAGAGAGAGGACCTGCTG"
                     /Tm="69.4"
                     /%GC="56.7"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /created_by="chenp"
                     /label="PC1658"
     primer_bind     complement(1879..1908)
                     /Sequence="CAGCAGGTCCTCTCTCTTCAGCTTCACGAG"
                     /Tm="69.4"
                     /%GC="56.7"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /created_by="chenp"
                     /label="PC1659"
     misc            1891..1893
                     /created_by="chenp"
                     /label="N394K"
     USER            1892..1904
                     /created_by="chenp"
     primer_bind     1892..1917
                     /Sequence="AGAGAGAGGACCTGCTGCGGAAGCAG"
                     /Tm="69.6"
                     /%GC="61.5"
                     /Hairpin_Tm="74.4"
                     /Self_Dimer_Tm="27.9"
                     /created_by="chenp"
                     /label="PC1648"
     misc_feature    1893
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     1945..1963
                     /Sequence="CACCAGATCCACCTGGGAG"
                     /%GC="63.2"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1501"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1x-cej84uk"">PC1501</a>"
                     /label="PC1501"
     primer_bind     2250..2274
                     /Sequence="GCTGTACGAGTACTTCACCGTGTAT"
                     /%GC="48.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ681"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:25-9qpay4m"">KZ681</a>"
                     /label="KZ681"
     primer_bind     2634..2655
                     /Sequence="CCTGTTCGACGACAAAGTGATG"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1502"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:22-cej84ul"">PC1502</a>"
                     /label="PC1502"
     primer_bind     2750..2774
                     /Sequence="TGGATTTCCTGAAGTCCGACGGCTT"
                     /%GC="52.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ682"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2a-9qpay4z"">KZ682</a>"
                     /label="KZ682"
     primer_bind     3208..3252
                     /Sequence="CTGTCCGACTACGATGTGGACGCTATCGTGCCTCAGAGCTTTCTG"
                     /created_by="Alexander Sousa"
                     /label="AVA216"
     primer_bind     complement(3208..3252)
                     /Sequence="CAGAAAGCTCTGAGGCACGATAGCGTCCACATCGTAGTCGGACAG"
                     /created_by="Alexander Sousa"
                     /label="AVA215"
     primer_bind     complement(3208..3231)
                     /Sequence="AGCGTCCACAUCGTAGTCGGACAG"
                     /%GC="58.3"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0893"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:b6-b14udud"">PC0893</a>"
                     /label="PC0893"
     primer_bind     3221..3250
                     /Sequence="ATGTGGACGCUATCGTGCCTCAGAGCTTTC"
                     /%GC="53.3"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0892"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:b1-b14udub"">PC0892</a>"
                     /label="PC0892"
     primer_bind     3250..3274
                     /Sequence="CTGAAGGACGACTCCATCGACAACA"
                     /%GC="52.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ683"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2f-9qpay56"">KZ683</a>"
                     /label="KZ683"
     primer_bind     3327..3348
                     /Sequence="CGAAGAGGTCGTGAAGAAGATG"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1503"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:27-cej84us"">PC1503</a>"
                     /label="PC1503"
     primer_bind     3750..3774
                     /Sequence="CAAGGTGTACGACGTGCGGAAGATG"
                     /%GC="56.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ684"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2k-9qpay5g"">KZ684</a>"
                     /label="KZ684"
     primer_bind     4014..4033
                     /Sequence="GACAGGCGGCTTCAGCAAAG"
                     /%GC="60.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1504"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2c-cej84ut"">PC1504</a>"
                     /label="PC1504"
     primer_bind     complement(4088..4112)
                     /Sequence="AAGCCGCCGUACTTCTTAGGGTCCC"
                     /%GC="60.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0895"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:bg-b14uduh"">PC0895</a>"
                     /label="PC0895"
     primer_bind     4250..4274
                     /Sequence="ACTTTCTGGAAGCCAAGGGCTACAA"
                     /%GC="48.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ685"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2p-9qpay5r"">KZ685</a>"
                     /label="KZ685"
     primer_bind     complement(4337..4359)
                     /Sequence="AGAGGCCAGCAUTCTCTTCCGGC"
                     /%GC="60.9"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0897"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:bq-b14uduk"">PC0897</a>"
                     /label="PC0897"
     primer_bind     complement(4581..4608)
                     /Sequence="ATCCCGGTGCTTGUTGTAGGCGGACAGC"
                     /%GC="60.7"
                     /note="Derived using Geneious Prime 2021.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1653"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:l3-d24gkh0"">PC1653</a>"
                     /label="PC1653"
     primer_bind     4595..4623
                     /Sequence="ACAAGCACCGGGAUAAGCCCATCAGAGAG"
                     /%GC="55.2"
                     /note="Derived using Geneious Prime 2021.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1652"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:ky-d24gkgz"">PC1652</a>"
                     /label="PC1652"
     primer_bind     4709..4728
                     /Sequence="GGAAGAGGTACACCAGCACC"
                     /%GC="60.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1505"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2h-cej84uv"">PC1505</a>"
                     /label="PC1505"
     primer_bind     4750..4774
                     /Sequence="CTGATCCACCAGAGCATCACCGGCC"
                     /%GC="64.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ686"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2u-9qpay5z"">KZ686</a>"
                     /label="KZ686"
     primer_bind     complement(4786..4812)
                     /Sequence="ACCTCCCAGCUGAGACAGGTCGATCCG"
                     /%GC="63.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1494"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:y-cej84u0"">PC1494</a>"
                     /label="PC1494"
     primer_bind     complement(4787..4805)
                     /Sequence="AGCTGAGACAGGUCGATCC"
                     /%GC="57.9"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0233"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1wx-998axgp"">PC0233</a>"
                     /label="PC0233"
     CDS             4816..4917
                     /created_by="chenp"
                     /label="SGGSx2-sv40NLS-SGGSx2"
     misc_feature    4818..4914
                     /Original_Bases="TGGAGGATCTAGCGGAGGATCCTCTGGCAGCGAGACACCAG
                     GAACAAGCGAGTCAGCAACACCAGAGAGCAGTGGCGGCAGCAGCGGCGGCAGC"
                     /label="TGGAGGATCTAGCGGAGGATCCTCTGGCAGCGAGACACCAGGAACAAGCG
                     AGTCAGCAACACCAGAGAGCAGTGGCGGCAGCAGCGGCGGCAGC"
                     /note="Geneious type: Editing History Replacement"
     CDS             4918..6948
                     /created_by="jessiedavis"
                     /label="RT codon optimized Genscript"
     primer_bind     5337..5356
                     /Sequence="CCCTTCTCACCAGTGGTATA"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="JRDSeq19"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:16w-cltirny"">JRDSeq19</a>"
                     /label="JRDSeq19"
     primer_bind     complement(6662..6680)
                     /Sequence="AAGGCATATCTGGAGTCTG"
                     /%GC="47.4"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="JRDSeq18"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:16r-cltirnv"">JRDSeq18</a>"
                     /label="JRDSeq18"
     CDS             6961..7014
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /Original_Translation="KRTADGSEFEPKKKRKV"
                     /label="SV40 NLS"
     misc_feature    6991..6993
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     primer_bind     complement(6994..7046)
                     /Sequence="AGCTTCACTCTCTUAGCGGCAGGGCCAGAGCCGACTTTCCTCTTCTT
                     CTTGGG"
                     /%GC="56.6"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1643"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:14h-d04t8ib"">PC1643</a>"
                     /label="PC1643"
     misc_feature    7015..7512
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     CDS             7024..7050
                     /created_by="chenp"
                     /label="cMyc-NLS"
     primer_bind     7033..7071
                     /Sequence="AAGAGAGTGAAGCUGGACGGATCCGGCGCAACAAACTTC"
                     /%GC="53.8"
                     /note="Derived using Geneious Prime 2021.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1642"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:14c-d04t8ia"">PC1642</a>"
                     /label="PC1642"
     USER            7038..>7050
                     /created_by="chenp"
     CDS             7060..7116
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding
                     separatepolypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             7117..>7512
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFT
                     GVNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHP
                     GIKAIVKDSDGQPTAVGIRELLPSGYVWEG"
                     /label="BSD"
     primer_bind     7514..7542
                     /Sequence="AACCGGTCATCAUCACCATCACCATTGAG"
                     /%GC="48.3"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1392"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:h0-cczyqfm"">PC1392</a>"
                     /label="PC1392"
     primer_bind     7514..7535
                     /Sequence="AACCGGTCATCAUCACCATCAC"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0192"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:34-96ures8"">PC0192</a>"
                     /label="PC0192"
     primer_bind     7515..7542
                     /Sequence="ACCGGTCATCAUCACCATCACCATTGAG"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1378"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:f2-cczyqes"">PC1378</a>"
                     /label="PC1378"
     primer_bind     7515..7532
                     /Sequence="ACCGGTCATCATCACCAT"
                     /Hairpin_Tm="32.9"
                     /Tm="55.5"
                     /Self_Dimer_Tm="None"
                     /%GC="50.0"
                     /created_by="primer3"
                     /label="JRD390"
     CDS             7521..7538
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /translation="HHHHHH"
                     /label="6xHis"
     primer_bind     complement(7551..7571)
                     /Sequence="CACAGTCGAGGCTGATCAGCG"
                     /%GC="61.9"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="BP0119 (pc3.1MCS-R)"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2u-93oy6a1"">BP0119 (pc3.1MCS-R)</a>"
                     /label="BP0119 (pc3.1MCS-R)"
     primer_bind     complement(7564..7584)
                     /Sequence="GGCAACTAGAAGGCACAGTCG"
                     /Hairpin_Tm="None"
                     /Tm="61.3"
                     /Self_Dimer_Tm="None"
                     /%GC="57.1"
                     /created_by="primer3"
                     /label="JRD371"
     polyA_signal    7567..7791
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     primer_bind     complement(7622..7646)
                     /Sequence="AGTGGGAGTGGCACCTTCCAGGGTC"
                     /%GC="64.0"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KZ687"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2z-9qpay63"">KZ687</a>"
                     /label="KZ687"
     misc_feature    7668
                     /label="mut"
     primer_bind     complement(7850..7870)
                     /Sequence="AGCTATGACCATGATUACGCC"
                     /%GC="47.6"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0004"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:t-92zzo16"">PC0004</a>"
                     /label="PC0004"
     primer_bind     7855..7876
                     /Sequence="AATCATGGTCATAGCUGTTTCC"
                     /%GC="40.9"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0002"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:j-92zzo0y"">PC0002</a>"
                     /label="PC0002"
     primer_bind     complement(7862..7878)
                     /%GC="47.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="49.1"
                     /Sequence="CAGGAAACAGCTATGAC"
                     /created_by="primer3"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="BP0006 (M13-Rev)"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:s-9439uh3"">BP0006 (M13-Rev)</a>"
                     /label="BP0006 (M13-Rev)"
     protein_bind    7886..7902
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7910..7940)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7955..7976
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the
                     presenceof cAMP."
                     /label="CAP binding site"
     primer_bind     complement(8063..8081)
                     /Sequence="GCGCCCAATACGCAAACCG"
                     /%GC="63.2"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0022"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:3b-92zzo2y"">PC0022</a>"
                     /label="PC0022"
     primer_bind     complement(8065..8085)
                     /Sequence="AAGAGCGCCCAAUACGCAAAC"
                     /%GC="52.4"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0187"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2f-96urern"">PC0187</a>"
                     /label="PC0187"
     primer_bind     complement(8168..8189)
                     /Sequence="ATCCCCTGATTCTGUGGATAAC"
                     /%GC="45.5"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0061"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8q-92zzo6v"">PC0061</a>"
                     /label="PC0061"
     primer_bind     8195..8217
                     /Sequence="AGGAAAGAACATGUGAGCAAAAG"
                     /%GC="39.1"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0060"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8l-92zzo6r"">PC0060</a>"
                     /label="PC0060"
     primer_bind     complement(8251..8272)
                     /Sequence="CTATGGAAAAACGCCAGCAACG"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC1010"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:7y-b64q19x"">PC1010</a>"
                     /label="PC1010"
     rep_origin      complement(8264..8852)
                     /direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     primer_bind     complement(8549..8577)
                     /Sequence="ACTCAAGACGAUAGTTACCGGATAAGGCG"
                     /%GC="48.3"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0970"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2e-b64q18g"">PC0970</a>"
                     /label="PC0970"
     primer_bind     8558..8590
                     /Sequence="CGGTAACTATCGTCTTGAGTCCAACCCGGTAAG"
                     /Tm="69.0"
                     /%GC="51.5"
                     /Hairpin_Tm="35.6"
                     /Self_Dimer_Tm="None"
                     /created_by="chenp"
                     /label="PC1660"
     primer_bind     complement(8558..8590)
                     /Sequence="CTTACCGGGTTGGACTCAAGACGATAGTTACCG"
                     /Tm="69.0"
                     /%GC="51.5"
                     /Hairpin_Tm="48.2"
                     /Self_Dimer_Tm="4.3"
                     /created_by="chenp"
                     /label="PC1661"
     primer_bind     8566..8592
                     /Sequence="ATCGTCTTGAGUCCAACCCGGTAAGAC"
                     /%GC="51.9"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0969"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:29-b64q18e"">PC0969</a>"
                     /label="PC0969"
     CDS             complement(9023..9883)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="AmpR"
     primer_bind     complement(9038..9058)
                     /Sequence="GCTGAGATAGGTGCCTCACTG"
                     /%GC="57.1"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0698"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:16y-a43x1kt"">PC0698</a>"
                     /label="PC0698"
     promoter        complement(9884..9988)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     10232..10252
                     /Sequence="CGATGTACGGGCCAGATATAC"
                     /%GC="52.4"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0037"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:5e-92zzo4g"">PC0037</a>"
                     /label="PC0037"
     primer_bind     10232..10252
                     /Sequence="CGATGTACGGGCCAGATATAC"
                     /%GC="52.4"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0037"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:5e-92zzo4g"">PC0037</a>"
                     /label="PC0037"
     primer_bind     complement(10242..6)
                     /Sequence="AATGTCAACGCGTAUATCTGGC"
                     /%GC="45.5"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0008"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1d-92zzo1h"">PC0008</a>"
                     /label="PC0008"
     primer_bind     10249..16
                     /Sequence="ATACGCGTTGACATUGATTATTGAC"
                     /%GC="36.0"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="PC0006"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:13-92zzo1c"">PC0006</a>"
                     /label="PC0006"
ORIGIN      
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcgacaag
      721 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
      781 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
      841 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
      901 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
      961 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     1021 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     1081 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     1141 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     1201 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     1261 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     1321 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag aaagctggaa
     1381 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     1441 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     1501 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     1561 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     1621 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     1681 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     1741 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     1801 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     1861 gacggcaccg aggaactgct cgtgaagctg aagagagagg acctgctgcg gaagcagcgg
     1921 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     1981 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     2041 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     2101 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     2161 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     2221 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     2281 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     2341 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     2401 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     2461 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     2521 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     2581 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     2641 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     2701 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     2761 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     2821 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     2881 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     2941 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     3001 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     3061 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     3121 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     3181 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct
     3241 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     3301 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     3361 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     3421 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     3481 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     3541 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     3601 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     3661 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     3721 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     3781 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     3841 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     3901 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     3961 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     4021 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     4081 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     4141 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     4201 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     4261 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     4321 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     4381 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     4441 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     4501 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     4561 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     4621 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     4681 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     4741 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     4801 cagctgggag gtgactccgg cggaagctct ggtggcagca agcggaccgc cgacggctct
     4861 gaattcgaga gccctaagaa gaaaagaaag gtgagcggag gctctagcgg cggaagcacc
     4921 ctgaacattg aagacgagta tagactgcat gaaacaagca aggaacccga cgtgtccctg
     4981 ggctccacct ggctgtccga ctttccccag gcctgggccg agacaggagg aatgggcctg
     5041 gccgtgcggc aggcacccct gatcatccct ctgaaggcca cctctacacc cgtgagcatc
     5101 aagcagtacc ctatgtctca ggaggccaga ctgggcatca agcctcacat ccagaggctg
     5161 ctggaccagg gcatcctggt gccatgccag agcccctgga acacaccact gctgcccgtg
     5221 aagaagccag gcaccaatga ctatagaccc gtgcaggatc tgagagaggt gaacaagagg
     5281 gtggaggata tccaccccac cgtgcccaac ccttacaatc tgctgtccgg cctgccccct
     5341 tctcaccagt ggtatacagt gctggacctg aaggatgcct tcttttgtct gagactgcac
     5401 cctaccagcc agccactgtt cgcctttgag tggagggacc ctgagatggg catctctggc
     5461 cagctgacct ggacacgcct gcctcagggc ttcaagaata gcccaacact gtttaacgag
     5521 gccctgcacc gcgacctggc agatttccgg atccagcacc cagatctgat cctgctgcag
     5581 tacgtggacg atctgctgct ggccgccacc agcgagctgg attgccagca gggaacacgc
     5641 gccctgctgc agaccctggg aaacctggga tatagggcat ccgccaagaa ggcccagatc
     5701 tgtcagaagc aggtgaagta cctgggctat ctgctgaagg agggccagag atggctgaca
     5761 gaggccagga aggagacagt gatgggccag ccaacaccca agaccccaag acagctgagg
     5821 gagttcctgg gcaaagcagg attttgcagg ctgttcatcc caggattcgc agagatggca
     5881 gcacctctgt acccactgac caagccgggc accctgttta attggggccc tgaccagcag
     5941 aaggcctatc aggagatcaa gcaggccctg ctgacagcac cagccctggg cctgccagac
     6001 ctgaccaagc ctttcgagct gtttgtggat gagaagcagg gctacgccaa gggcgtgctg
     6061 acccagaagc tgggaccatg gagacggccc gtggcctatc tgtccaagaa gctggaccca
     6121 gtggcagcag gatggccacc atgcctgagg atggtggcag caatcgccgt gctgacaaag
     6181 gatgccggca agctgaccat gggacagcca ctggtcatcc tggcaccaca cgcagtggag
     6241 gccctggtga agcagcctcc agatcgctgg ctgtctaacg cccggatgac acactaccag
     6301 gccctgctgc tggacaccga tcgcgtgcag tttggccctg tggtggccct gaatccagcc
     6361 accctgctgc ctctgccaga ggagggcctg cagcacaact gtctggacat cctggcagag
     6421 gcacacggaa caaggccaga cctgaccgat cagcccctgc ctgacgccga tcacacatgg
     6481 tataccgatg gaagctccct gctgcaggag ggccagagga aggcaggagc agcagtgacc
     6541 acagagacag aagtgatctg ggccaaggcc ctgccagcag gcacatccgc ccagcgggcc
     6601 gagctgatcg ccctgaccca ggccctgaag atggccgagg gcaagaagct gaacgtgtac
     6661 acagactcca gatatgcctt cgccaccgca cacatccacg gagagatcta caggcgccgg
     6721 ggctggctga cctctgaggg caaggagatc aagaacaagg atgagatcct ggccctgctg
     6781 aaggccctgt ttctgcccaa gcggctgagc atcatccact gtcctggaca ccagaaggga
     6841 cactccgccg aggcaagggg caatcggatg gccgaccagg ccgccagaaa ggctgctatt
     6901 actgaaactc ccgacacttc cactctgctg attgaaaact cctccccttc tggcggctca
     6961 aaaagaaccg ccgacggcag cgaattcgag tctcccaaga agaagaggaa agtcggctct
     7021 ggccctgccg ctaagagagt gaagctggac ggatccggcg caacaaactt ctctctgctg
     7081 aaacaagccg gagatgtcga agagaatcct ggaccgatgg ccaagccttt gtctcaagaa
     7141 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     7201 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     7261 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     7321 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     7381 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     7441 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     7501 gtgtgggagg gctaaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     7561 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     7621 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgagaaa attgcatcgc
     7681 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     7741 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     7801 cggaaagaac cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat
     7861 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     7921 ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg
     7981 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     8041 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     8101 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     8161 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     8221 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     8281 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     8341 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     8401 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8461 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8521 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8581 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8641 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8701 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8761 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     8821 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     8881 ctgacactca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     8941 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     9001 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     9061 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     9121 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     9181 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     9241 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     9301 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     9361 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     9421 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     9481 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     9541 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     9601 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     9661 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     9721 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     9781 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     9841 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     9901 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     9961 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
    10021 acggatcggg agatcgatct cccgatcccc tagggtctac tctcagtaca atctgctctg
    10081 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
    10141 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
    10201 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
