LOCUS       pTBL2670_MTK3_PE        8058 bp    DNA        circular     09-AUG-2024
DEFINITION  .
ACCESSION   <unknown id>                                                       
VERSION     <unknown id>                                                       
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     misc_feature    606..609
                     /locus_tag="BsaI"
                     /label="BsaI"
                     /ApEinfo_label="BsaI"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    complement(7003..7005)
                     /locus_tag="BsaI(1)"
                     /label="BsaI(1)"
                     /ApEinfo_label="BsaI"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    599..604
                     /locus_tag="BsaI(2)"
                     /label="BsaI(2)"
                     /ApEinfo_label="BsaI"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    complement(7007..7012)
                     /locus_tag="BsaI(3)"
                     /label="BsaI(3)"
                     /ApEinfo_label="BsaI"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    complement(join(7889..8058,1..594))
                     /locus_tag="ColE1"
                     /label="ColE1"
                     /ApEinfo_label="ColE1"
                     /ApEinfo_fwdcolor="#7f7f7f"
                     /ApEinfo_revcolor="#7f7f7f"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    complement(7784..7888)
                     /locus_tag="CamR Promoter"
                     /label="CamR Promoter"
                     /ApEinfo_label="CamR Promoter"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    complement(7124..7783)
                     /locus_tag="CamR"
                     /label="CamR"
                     /ApEinfo_label="CamR"
                     /ApEinfo_fwdcolor="#0000ff"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    complement(7015..7123)
                     /locus_tag="CamR Terminator"
                     /label="CamR Terminator"
                     /ApEinfo_label="CamR Terminator"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     610..625
                     /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>"
                     /locus_tag="PC0198"
                     /label="PC0198"
                     /ApEinfo_label="PC0198"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             664..4764
                     /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"
                     /locus_tag="Cas9(R221K N394K H840A)"
                     /label="Cas9(R221K N394K H840A)"
                     /ApEinfo_label="Cas9(R221K N394K H840A)"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     679..696
                     /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>"
                     /locus_tag="PC0293"
                     /label="PC0293"
                     /ApEinfo_label="PC0293"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(1045..1064)
                     /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>"
                     /locus_tag="PC0229"
                     /label="PC0229"
                     /ApEinfo_label="PC0229"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(1144..1163)
                     /Sequence="AAGTGGCCCCGGAACTTGAT"
                     /Hairpin_Tm="33.0"
                     /Tm="61.8"
                     /Self_Dimer_Tm="0.7"
                     /%GC="55.0"
                     /created_by="primer3"
                     /locus_tag="PC0873"
                     /label="PC0873"
                     /ApEinfo_label="PC0873"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(1294..1325)
                     /Sequence="AGCTTTCTGCTCTTGCTCAGTCTGGCAGACAG"
                     /Tm="71.1"
                     /%GC="53.1"
                     /Hairpin_Tm="52.7"
                     /Self_Dimer_Tm="40.1"
                     /created_by="chenp"
                     /locus_tag="PC1651"
                     /label="PC1651"
                     /ApEinfo_label="PC1651"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1321..1322
                     /Original_Bases="CG"
                     /note="Geneious type: Editing History Replacement"
                     /locus_tag="CG"
                     /label="CG"
                     /ApEinfo_label="CG"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(1828..1857)
                     /Sequence="CAGCAGGTCCTCTCTCTTCAGCTTCACGAG"
                     /Tm="69.4"
                     /%GC="56.7"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /created_by="chenp"
                     /locus_tag="PC1659"
                     /label="PC1659"
                     /ApEinfo_label="PC1659"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    1842..1842
                     /Original_Bases="C"
                     /note="Geneious type: Editing History Replacement"
                     /locus_tag="C"
                     /label="C"
                     /ApEinfo_label="C"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     2699..2723
                     /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>"
                     /locus_tag="KZ682"
                     /label="KZ682"
                     /ApEinfo_label="KZ682"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     3170..3199
                     /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>"
                     /locus_tag="PC0892"
                     /label="PC0892"
                     /ApEinfo_label="PC0892"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     3963..3982
                     /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>"
                     /locus_tag="PC1504"
                     /label="PC1504"
                     /ApEinfo_label="PC1504"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(4530..4557)
                     /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>"
                     /locus_tag="PC1653"
                     /label="PC1653"
                     /ApEinfo_label="PC1653"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(4735..4761)
                     /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>"
                     /locus_tag="PC1494"
                     /label="PC1494"
                     /ApEinfo_label="PC1494"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             4867..6897
                     /created_by="jessiedavis"
                     /locus_tag="RT codon optimized Genscript"
                     /label="RT codon optimized Genscript"
                     /ApEinfo_label="RT codon optimized Genscript"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    6940..6942
                     /Original_Bases="SV40 NLS"
                     /note="Geneious type: Editing History Insertion"
                     /locus_tag="SV40 NLS"
                     /label="SV40 NLS"
                     /ApEinfo_label="SV40 NLS"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             610..663
                     /codon_start="1"
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="MKRTADGSEFESPKKKRKV"
                     /locus_tag="SV40 NLS(1)"
                     /label="SV40 NLS(1)"
                     /ApEinfo_label="SV40 NLS"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     664..684
                     /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>"
                     /locus_tag="PC1498"
                     /label="PC1498"
                     /ApEinfo_label="PC1498"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     699..723
                     /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>"
                     /locus_tag="KZ678"
                     /label="KZ678"
                     /ApEinfo_label="KZ678"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1052..1071
                     /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>"
                     /locus_tag="PC0228"
                     /label="PC0228"
                     /ApEinfo_label="PC0228"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(1144..1163)
                     /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>"
                     /locus_tag="PC0873(1)"
                     /label="PC0873(1)"
                     /ApEinfo_label="PC0873"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     USER            1313..1325
                     /created_by="chenp"
                     /locus_tag="USER"
                     /label="USER"
                     /ApEinfo_label="USER"
                     /ApEinfo_fwdcolor="#00DF87"
                     /ApEinfo_revcolor="#00DF87"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1699..1723
                     /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>"
                     /locus_tag="KZ680"
                     /label="KZ680"
                     /ApEinfo_label="KZ680"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc            1840..1842
                     /created_by="chenp"
                     /locus_tag="N394K"
                     /label="N394K"
                     /ApEinfo_label="N394K"
                     /ApEinfo_fwdcolor="#008065"
                     /ApEinfo_revcolor="#008065"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1894..1912
                     /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>"
                     /locus_tag="PC1501"
                     /label="PC1501"
                     /ApEinfo_label="PC1501"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     3157..3201
                     /Sequence="CTGTCCGACTACGATGTGGACGCTATCGTGCCTCAGAGCTTTCTG"
                     /created_by="Alexander Sousa"
                     /locus_tag="AVA216"
                     /label="AVA216"
                     /ApEinfo_label="AVA216"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     3199..3223
                     /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>"
                     /locus_tag="KZ683"
                     /label="KZ683"
                     /ApEinfo_label="KZ683"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(4037..4061)
                     /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>"
                     /locus_tag="PC0895"
                     /label="PC0895"
                     /ApEinfo_label="PC0895"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     4544..4572
                     /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>"
                     /locus_tag="PC1652"
                     /label="PC1652"
                     /ApEinfo_label="PC1652"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(4736..4754)
                     /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>"
                     /locus_tag="PC0233"
                     /label="PC0233"
                     /ApEinfo_label="PC0233"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     5286..5305
                     /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>"
                     /locus_tag="JRDSeq19"
                     /label="JRDSeq19"
                     /ApEinfo_label="JRDSeq19"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    6964..7001
                     /Original_Bases="Geneious type: Editing History Insertion"
                     /note="Geneious type: Editing History Insertion"
                     /locus_tag="Geneious type: Editing History Insertion"
                     /label="Geneious type: Editing History Insertion"
                     /ApEinfo_label="Geneious type: Editing History Insertion"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     623..655
                     /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>"
                     /locus_tag="PC0738"
                     /label="PC0738"
                     /ApEinfo_label="PC0738"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(671..692)
                     /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>"
                     /locus_tag="PC0292"
                     /label="PC0292"
                     /ApEinfo_label="PC0292"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(742..763)
                     /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>"
                     /locus_tag="PC1499"
                     /label="PC1499"
                     /ApEinfo_label="PC1499"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1081..1100
                     /Sequence="AAACTGGTGGACAGCACCGA"
                     /Hairpin_Tm="51.8"
                     /Tm="62.0"
                     /Self_Dimer_Tm="None"
                     /%GC="55.0"
                     /created_by="primer3"
                     /locus_tag="PC0872"
                     /label="PC0872"
                     /ApEinfo_label="PC0872"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1198..1218
                     /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>"
                     /locus_tag="PC1500"
                     /label="PC1500"
                     /ApEinfo_label="PC1500"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1313..1345
                     /Sequence="AGAGCAGAAAGCTGGAAAATCTGATCGCCCAGC"
                     /Tm="71.5"
                     /%GC="51.5"
                     /Hairpin_Tm="58.2"
                     /Self_Dimer_Tm="29.7"
                     /created_by="chenp"
                     /locus_tag="PC1650"
                     /label="PC1650"
                     /ApEinfo_label="PC1650"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(1821..1853)
                     /Sequence="AGGTCCTCTCTCTTCAGCTTCACGAGCAGTTCC"
                     /Tm="71.4"
                     /%GC="54.5"
                     /Hairpin_Tm="54.3"
                     /Self_Dimer_Tm="0.5"
                     /created_by="chenp"
                     /locus_tag="PC1649"
                     /label="PC1649"
                     /ApEinfo_label="PC1649"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     USER            1841..1853
                     /created_by="chenp"
                     /locus_tag="USER(1)"
                     /label="USER(1)"
                     /ApEinfo_label="USER"
                     /ApEinfo_fwdcolor="#13AAFE"
                     /ApEinfo_revcolor="#13AAFE"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     2199..2223
                     /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>"
                     /locus_tag="KZ681"
                     /label="KZ681"
                     /ApEinfo_label="KZ681"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(3157..3201)
                     /Sequence="CAGAAAGCTCTGAGGCACGATAGCGTCCACATCGTAGTCGGACAG"
                     /created_by="Alexander Sousa"
                     /locus_tag="AVA215"
                     /label="AVA215"
                     /ApEinfo_label="AVA215"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     3276..3297
                     /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>"
                     /locus_tag="PC1503"
                     /label="PC1503"
                     /ApEinfo_label="PC1503"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     4199..4223
                     /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>"
                     /locus_tag="KZ685"
                     /label="KZ685"
                     /ApEinfo_label="KZ685"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     4658..4677
                     /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>"
                     /locus_tag="PC1505"
                     /label="PC1505"
                     /ApEinfo_label="PC1505"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             4765..4866
                     /created_by="chenp"
                     /locus_tag="SGGSx2-sv40NLS-SGGSx2"
                     /label="SGGSx2-sv40NLS-SGGSx2"
                     /ApEinfo_label="SGGSx2-sv40NLS-SGGSx2"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(6611..6629)
                     /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>"
                     /locus_tag="JRDSeq18"
                     /label="JRDSeq18"
                     /ApEinfo_label="JRDSeq18"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             6973..7001
                     /created_by="chenp"
                     /locus_tag="c-Myc NLS"
                     /label="c-Myc NLS"
                     /ApEinfo_label="c-Myc NLS"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(610..640)
                     /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>"
                     /locus_tag="PC0234"
                     /label="PC0234"
                     /ApEinfo_label="PC0234"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(633..662)
                     /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>"
                     /locus_tag="PC0831"
                     /label="PC0831"
                     /ApEinfo_label="PC0831"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     674..698
                     /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>"
                     /locus_tag="PC1335"
                     /label="PC1335"
                     /ApEinfo_label="PC1335"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(803..827)
                     /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>"
                     /locus_tag="KZ677"
                     /label="KZ677"
                     /ApEinfo_label="KZ677"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1081..1100
                     /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>"
                     /locus_tag="PC0872(1)"
                     /label="PC0872(1)"
                     /ApEinfo_label="PC0872"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1199..1223
                     /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>"
                     /locus_tag="KZ679"
                     /label="KZ679"
                     /ApEinfo_label="KZ679"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc            1321..1323
                     /created_by="chenp"
                     /modified_by="chenp"
                     /locus_tag="R221K"
                     /label="R221K"
                     /ApEinfo_label="R221K"
                     /ApEinfo_fwdcolor="#00D95A"
                     /ApEinfo_revcolor="#00D95A"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1828..1857
                     /Sequence="CTCGTGAAGCTGAAGAGAGAGGACCTGCTG"
                     /Tm="69.4"
                     /%GC="56.7"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /created_by="chenp"
                     /locus_tag="PC1658"
                     /label="PC1658"
                     /ApEinfo_label="PC1658"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     1841..1866
                     /Sequence="AGAGAGAGGACCTGCTGCGGAAGCAG"
                     /Tm="69.6"
                     /%GC="61.5"
                     /Hairpin_Tm="74.4"
                     /Self_Dimer_Tm="27.9"
                     /created_by="chenp"
                     /locus_tag="PC1648"
                     /label="PC1648"
                     /ApEinfo_label="PC1648"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     2583..2604
                     /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>"
                     /locus_tag="PC1502"
                     /label="PC1502"
                     /ApEinfo_label="PC1502"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(3157..3180)
                     /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>"
                     /locus_tag="PC0893"
                     /label="PC0893"
                     /ApEinfo_label="PC0893"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     3699..3723
                     /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>"
                     /locus_tag="KZ684"
                     /label="KZ684"
                     /ApEinfo_label="KZ684"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     complement(4286..4308)
                     /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>"
                     /locus_tag="PC0897"
                     /label="PC0897"
                     /ApEinfo_label="PC0897"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     primer_bind     4699..4723
                     /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>"
                     /locus_tag="KZ686"
                     /label="KZ686"
                     /ApEinfo_label="KZ686"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    4767..4863
                     /Original_Bases="TGGAGGATCTAGCGGAGGATCCTCTGGCAGCGAGACACCAG
                     GAACAAGCGAGTCAGCAACACCAGAGAGCAGTGGCGGCAGCAGCGGCGGCAGC"
                     /note="Geneious type: Editing History Replacement"
                     /locus_tag="TGGAGGATCTAGCGGAGGATCCTCTGGCAGCGAGACACCAGG
                     AACAAGCG AGTCAGCAACACCAGAGAGCAGTGGCGGCAGCAGCGGCGGCAGC"
                     /label="TGGAGGATCTAGCGGAGGATCCTCTGGCAGCGAGACACCAGG
                     AACAAGCG AGTCAGCAACACCAGAGAGCAGTGGCGGCAGCAGCGGCGGCAGC"
                     /ApEinfo_label="TGGAGGATCTAGCGGAGGATCCTCTGGCAGCGAGACACCAGG
                     AACAAGCG AGTCAGCAACACCAGAGAGCAGTGGCGGCAGCAGCGGCGGCAGC"
                     /ApEinfo_fwdcolor="#7eff74"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     CDS             6910..6963
                     /codon_start="1"
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /Original_Translation="KRTADGSEFEPKKKRKV"
                     /locus_tag="SV40 NLS(2)"
                     /label="SV40 NLS(2)"
                     /ApEinfo_label="SV40 NLS"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     USER            6987..7001
                     /created_by="chenp"
                     /locus_tag="USER(2)"
                     /label="USER(2)"
                     /ApEinfo_label="USER"
                     /ApEinfo_fwdcolor="#3A93FE"
                     /ApEinfo_revcolor="#3A93FE"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
     misc_feature    6030..6035
                     /locus_tag="Esp3I_site_changed"
                     /label="Esp3I_site_changed"
                     /ApEinfo_label="Esp3I_site_changed"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="cyan"
                     /ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
                     -0.5} {} 0} width 5 offset 0"
ORIGIN
        1 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
       61 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
      121 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      181 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      241 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      301 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      361 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      421 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      481 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      541 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtagtcgg
      601 tctcatatga aacggacagc cgacggaagc gagttcgagt caccaaagaa gaagcggaaa
      661 gtcgacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg
      721 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
      781 cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag
      841 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
      901 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
      961 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     1021 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     1081 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     1141 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     1201 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     1261 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     1321 aagctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac
     1381 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     1441 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     1501 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     1561 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     1621 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     1681 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     1741 ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     1801 gaaaagatgg acggcaccga ggaactgctc gtgaagctga agagagagga cctgctgcgg
     1861 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     1921 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     1981 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     2041 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     2101 gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     2161 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     2221 tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     2281 agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     2341 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     2401 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     2461 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     2521 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     2581 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     2641 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     2701 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     2761 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     2821 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     2881 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     2941 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     3001 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     3061 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     3121 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggacgct
     3181 atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc
     3241 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     3301 aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg
     3361 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     3421 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     3481 actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc
     3541 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     3601 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     3661 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     3721 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     3781 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     3841 cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt
     3901 gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     3961 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc
     4021 gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc
     4081 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
     4141 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
     4201 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
     4261 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg
     4321 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
     4381 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
     4441 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
     4501 atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag
     4561 cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
     4621 cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa
     4681 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
     4741 gacctgtctc agctgggagg tgactccggc ggaagctctg gtggcagcaa gcggaccgcc
     4801 gacggctctg aattcgagag ccctaagaag aaaagaaagg tgagcggagg ctctagcggc
     4861 ggaagcaccc tgaacattga agacgagtat agactgcatg aaacaagcaa ggaacccgac
     4921 gtgtccctgg gctccacctg gctgtccgac tttccccagg cctgggccga gacaggagga
     4981 atgggcctgg ccgtgcggca ggcacccctg atcatccctc tgaaggccac ctctacaccc
     5041 gtgagcatca agcagtaccc tatgtctcag gaggccagac tgggcatcaa gcctcacatc
     5101 cagaggctgc tggaccaggg catcctggtg ccatgccaga gcccctggaa cacaccactg
     5161 ctgcccgtga agaagccagg caccaatgac tatagacccg tgcaggatct gagagaggtg
     5221 aacaagaggg tggaggatat ccaccccacc gtgcccaacc cttacaatct gctgtccggc
     5281 ctgccccctt ctcaccagtg gtatacagtg ctggacctga aggatgcctt cttttgtctg
     5341 agactgcacc ctaccagcca gccactgttc gcctttgagt ggagggaccc tgagatgggc
     5401 atctctggcc agctgacctg gacacgcctg cctcagggct tcaagaatag cccaacactg
     5461 tttaacgagg ccctgcaccg cgacctggca gatttccgga tccagcaccc agatctgatc
     5521 ctgctgcagt acgtggacga tctgctgctg gccgccacca gcgagctgga ttgccagcag
     5581 ggaacacgcg ccctgctgca gaccctggga aacctgggat atagggcatc cgccaagaag
     5641 gcccagatct gtcagaagca ggtgaagtac ctgggctatc tgctgaagga gggccagaga
     5701 tggctgacag aggccaggaa ggagacagtg atgggccagc caacacccaa gaccccaaga
     5761 cagctgaggg agttcctggg caaagcagga ttttgcaggc tgttcatccc aggattcgca
     5821 gagatggcag cacctctgta cccactgacc aagccgggca ccctgtttaa ttggggccct
     5881 gaccagcaga aggcctatca ggagatcaag caggccctgc tgacagcacc agccctgggc
     5941 ctgccagacc tgaccaagcc tttcgagctg tttgtggatg agaagcaggg ctacgccaag
     6001 ggcgtgctga cccagaagct gggaccatgg agGcggcccg tggcctatct gtccaagaag
     6061 ctggacccag tggcagcagg atggccacca tgcctgagga tggtggcagc aatcgccgtg
     6121 ctgacaaagg atgccggcaa gctgaccatg ggacagccac tggtcatcct ggcaccacac
     6181 gcagtggagg ccctggtgaa gcagcctcca gatcgctggc tgtctaacgc ccggatgaca
     6241 cactaccagg ccctgctgct ggacaccgat cgcgtgcagt ttggccctgt ggtggccctg
     6301 aatccagcca ccctgctgcc tctgccagag gagggcctgc agcacaactg tctggacatc
     6361 ctggcagagg cacacggaac aaggccagac ctgaccgatc agcccctgcc tgacgccgat
     6421 cacacatggt ataccgatgg aagctccctg ctgcaggagg gccagaggaa ggcaggagca
     6481 gcagtgacca cagagacaga agtgatctgg gccaaggccc tgccagcagg cacatccgcc
     6541 cagcgggccg agctgatcgc cctgacccag gccctgaaga tggccgaggg caagaagctg
     6601 aacgtgtaca cagactccag atatgccttc gccaccgcac acatccacgg agagatctac
     6661 aggcgccggg gctggctgac ctctgagggc aaggagatca agaacaagga tgagatcctg
     6721 gccctgctga aggccctgtt tctgcccaag cggctgagca tcatccactg tcctggacac
     6781 cagaagggac actccgccga ggcaaggggc aatcggatgg ccgaccaggc cgccagaaag
     6841 gctgctatta ctgaaactcc cgacacttcc actctgctga ttgaaaactc ctccccttct
     6901 ggcggctcaa aaagaaccgc cgacggcagc gaattcgagt ctcccaagaa gaagaggaaa
     6961 gtcggctctg gccctgccgc taagagagtg aagctggacg gatcctgaga ccagaccaat
     7021 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     7081 tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc
     7141 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     7201 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     7261 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     7321 tggtgaaact cacccaggga ttggctgaaa cgaaaaacat attctcaata aaccctttag
     7381 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     7441 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     7501 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     7561 tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     7621 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     7681 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     7741 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     7801 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     7861 agttggaacc tcttacgtgc ccgatcaatc atgaccaaaa tcccttaacg tgagttttcg
     7921 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7981 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     8041 ccggatcaag agctacca    
//
