LOCUS       eeBxb1_IVT              4984 bp    DNA     circular UNA 04-JUN-2024
DEFINITION  [pBR322, Cb] pCMV-SpPE2 mRNA Plasmid for IVT.
ACCESSION   urn.local...1h-hsvfz4k
VERSION     urn.local...1h-hsvfz4k
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     enhancer        1..380
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     primer_bind     complement(67..86)
                     /Sequence="TTATGTAACGCGGAACTCCA"
                     /%GC="45.0"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB1110"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1sh-c6uht7h"">QB1110</a>"
                     /label="QB1110"
     promoter        381..584
                     /note="human cytomegalovirus (CMV) immediate
                     early<br>promoter"
                     /label="CMV promoter"
     primer_bind     535..555
                     /note="Human CMV immediate early promoter, forward primer"
                     /label="CMV-F"
     primer_bind     535..555
                     /Sequence="CGCAAATGGGCGGTAGGCGTG"
                     /%GC="66.7"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0012"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1bo-c6uht51"">QB0012</a>"
                     /label="QB0012"
     primer_bind     535..555
                     /Sequence="CGCAAATGGGCGGTAGGCGTG"
                     /%GC="66.7"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1753"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:3ax-f3up2sk"">oSP1753</a>"
                     /label="oSP1753"
     primer_bind     542..563
                     /Sequence="GGGCGGTAGGCGTGTACGGTGG"
                     /%GC="72.7"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0976"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1s2-c6uht7e"">QB0976</a>"
                     /label="QB0976"
     regulatory      complement(583..605)
                     /note="/created_by=everette"
                     /label="VECTOR PRIMER 1"
     regulatory      complement(606..661)
                     /note="/created_by=everette"
                     /label="VECTOR PRIMER 1"
     misc_feature    606..677
                     /note="/Original_Bases=CGCTAGAGATCCGCGGCCGCTAATACGACTCACTA
                     TAGGGA<br>GAGCC"
                     /label="CGCTAGAGATCCGCGGCCGCTAATACGACTCACTATAGGGAGAGCC"
                     /note="Geneious type: Editing History Replacement"
     misc_RNA        620..683
                     /note="/created_by=nicolegaudelli"
                     /label="5'UTR sequence"
     promoter        620..639
                     /note="/created_by=nicolegaudelli"
                     /label="T7 promoter with sequence mismatch"
     primer_bind     complement(650..683)
                     /Sequence="GGTGGCTCTTATATTTCTTCTTACTCTTCTTTTC"
                     /%GC="35.3"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP2004"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:wx-fh0vjsu"">oSP2004</a>"
                     /label="oSP2004"
     regulatory      652..683
                     /note="/created_by=everette"
                     /label="EDITOR PRIMER 1"
     CDS             684..2183
                     /created_by="smreets"
                     /label="B.400.2"
     ORF             <2184..2186
                     /codon_start=1
                     /note="/created_by=User"
                     /label="BxB1 CO"
     regulatory      complement(2187..2189)
                     /note="/created_by=everette"
                     /label="EDITOR PRIMER 2"
     primer_bind     2187..2213
                     /Sequence="TAATTAATTAAGCTGCCTTCTGCGGGG"
                     /%GC="44.4"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP2005"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:wy-fh0vjsu"">oSP2005</a>"
                     /label="oSP2005"
     regulatory      complement(2190..2240)
                     /note="/created_by=everette"
                     /note="/modified_by=everette"
                     /label="EDITOR PRIMER 2"
     misc_feature    2190..2290
                     /note="/Original_Bases=CCGGTCATCATCACCATCACCATTGAGTTTAAACC
                     CGCTGA<br>TCAGCCT"
                     /label="CCGGTCATCATCACCATCACCATTGAGTTTAAACCCGCTGATCAGCCT"
                     /note="Geneious type: Editing History Replacement"
     regulatory      2228..2290
                     /note="/created_by=everette"
                     /label="VECTOR PRIMER 2"
     primer_bind     complement(2262..2290)
                     /Sequence="CTTCCTACTCAGGCTTTATTCAAAGACCA"
                     /%GC="41.4"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1987"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:ub-fh0vjs8"">oSP1987</a>"
                     /label="oSP1987"
     regulatory      2291..2311
                     /note="/created_by=everette"
                     /label="VECTOR PRIMER 2"
     polyA_signal    2294..2518
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     primer_bind     complement(2355..2374)
                     /Sequence="CAGTGGGAGTGGCACCTTCC"
                     /%GC="65.0"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1047"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1d-dm5dbes"">oSP1047</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:49x-dll8epl"">oSP1047</a>"
                     /label="oSP1047"
     primer_bind     complement(2589..2605)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     primer_bind     complement(2589..2605)
                     /note="In lacZ gene. Also called M13-rev"
                     /label="M13 Reverse"
     primer_bind     complement(2589..2605)
                     /Sequence="CAGGAAACAGCTATGAC"
                     /%GC="47.1"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0048"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1go-c6uht5t"">QB0048</a>"
                     /label="QB0048"
     primer_bind     complement(2602..2625)
                     /Sequence="GAGCGGATAACAATTTCACACAGG"
                     /%GC="45.8"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0108"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1p5-c6uht6y"">QB0108</a>"
                     /label="QB0108"
     primer_bind     complement(2602..2624)
                     /note="In lacZ gene"
                     /label="M13/pUC Reverse"
     primer_bind     complement(2602..2624)
                     /Sequence="AGCGGATAACAATTTCACACAGG"
                     /%GC="43.5"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0050"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1gy-c6uht5u"">QB0050</a>"
                     /label="QB0050"
     protein_bind    2613..2629
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator
                     to<br>inhibit transcription in E. coli. This inhibition
                     can be relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(2637..2667)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     primer_bind     2674..2724
                     /Sequence="GGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACT
                     GCCC"
                     /%GC="51.0"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1171"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:17g-drqs4cv"">oSP1171</a>"
                     /label="oSP1171"
     primer_bind     complement(2678..2699)
                     /Sequence="GTGAGTTAGCTCACTCATTAGG"
                     /%GC="45.5"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP495"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:avg-dkzgvgi"">oSP495</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:7ov-dkzgu8r"">oSP495</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:9i4-cj5pty4"">oSP495</a>"
                     /label="oSP495"
     protein_bind    2682..2703
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the
                     presence<br>of cAMP."
                     /label="CAP binding site"
     primer_bind     2743..2765
                     /Sequence="CTGTCGTGCCAGCTGCATTAATG"
                     /%GC="52.2"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP865"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:c8r-dkzgvjj"">oSP865</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:926-dkzgud0"">oSP865</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2qw-dck6bx6"">oSP865</a>"
                     /label="oSP865"
     primer_bind     complement(2773..2790)
                     /Sequence="GCCTCTCCCCGCGCGTTG"
                     /%GC="77.8"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1214"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6h8-dvtyyoa"">oSP1214</a>"
                     /label="oSP1214"
     primer_bind     complement(2789..2809)
                     /Sequence="AGCGCCCAATACGCAAACCGC"
                     /%GC="61.9"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1107"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2bl-dn4yt2z"">oSP1107</a>"
                     /label="oSP1107"
     primer_bind     2810..2830
                     /Sequence="CTTCCGCTTCCTCGCTCACTG"
                     /%GC="61.9"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1109"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2bv-dn4yt32"">oSP1109</a>"
                     /label="oSP1109"
     primer_bind     complement(2819..2837)
                     /Sequence="AGCGAGTCAGTGAGCGAGG"
                     /%GC="63.2"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0039"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1ff-c6uht5n"">QB0039</a>"
                     /label="QB0039"
     primer_bind     complement(2820..2837)
                     /note="L4440 vector, forward primer"
                     /label="L4440"
     primer_bind     complement(2845..2893)
                     /Sequence="GTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACG
                     AC"
                     /%GC="59.2"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1172"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:17l-drqs4cw"">oSP1172</a>"
                     /label="oSP1172"
     primer_bind     complement(2863..2884)
                     /Sequence="CCTTTGAGTGAGCTGATACCGC"
                     /%GC="54.5"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP827"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8ww-dkzgucl"">oSP827</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:uq-dc0szt5"">oSP827</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:c3h-dkzgvj7"">oSP827</a>"
                     /label="oSP827"
     primer_bind     2864..2885
                     /Sequence="CGGTATCAGCTCACTCAAAGGC"
                     /%GC="54.5"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1170"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:17b-drqs4ct"">oSP1170</a>"
                     /label="oSP1170"
     primer_bind     2888..2914
                     /Sequence="TAATACGGTTATCCACAGAATCAGGGG"
                     /%GC="44.4"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1487"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:38s-eh5sytc"">oSP1487</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2vq-eh5sxfu"">oSP1487</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:10a-ebp5bd0"">oSP1487</a>"
                     /label="oSP1487"
     rep_origin      complement(2991..3579)
                     /snapgene_direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     primer_bind     complement(3005..3022)
                     /Sequence="GATGCTCGTCAGGGGGGC"
                     /%GC="72.2"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP301"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:32i-cf0ayl5"">oSP301</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:a4i-dkzgvet"">oSP301</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6xx-dkzgu5v"">oSP301</a>"
                     /label="oSP301"
     primer_bind     complement(3051..3078)
                     /Sequence="GTATCTTTATAGTCCTGTCGGGTTTCGC"
                     /%GC="46.4"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP785"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8r2-dkzguc2"">oSP785</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:bxn-dkzgviu"">oSP785</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6k3-dcvoynx"">oSP785</a>"
                     /label="oSP785"
     primer_bind     complement(3071..3090)
                     /note="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     primer_bind     complement(3092..3109)
                     /Sequence="GCACGAGGGAGCTTCCAG"
                     /%GC="66.7"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="Ju1-R2"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1v9-c6uhujv"">Ju1-R2</a>"
                     /label="Ju1-R2"
     primer_bind     3214..3232
                     /Sequence="GTGTAGGTCGTTCGCTCCA"
                     /%GC="57.9"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0766"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1rs-c6uht7c"">QB0766</a>"
                     /label="QB0766"
     primer_bind     3331..3349
                     /Sequence="CCACTGGCAGCAGCCACTG"
                     /%GC="68.4"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1195"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:z5-ds50lrl"">oSP1195</a>"
                     /label="oSP1195"
     primer_bind     complement(3339..3358)
                     /Sequence="TCCTGTTACCAGTGGCTGCT"
                     /%GC="55.0"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0767"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1rx-c6uht7d"">QB0767</a>"
                     /label="QB0767"
     CDS             complement(3750..4610)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin,
                     carbenicillin,<br>and related antibiotics"
                     /product="beta-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="AmpR"
     primer_bind     complement(3796..3816)
                     /Sequence="GGCAACTATGGATGAACGAAA"
                     /%GC="42.9"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="Ju1-R1"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1v4-c6uhuju"">Ju1-R1</a>"
                     /label="Ju1-R1"
     primer_bind     complement(3796..3816)
                     /Sequence="GGCAACTATGGATGAACGAAA"
                     /%GC="42.9"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1216"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6hi-dvtyyoc"">oSP1216</a>"
                     /label="oSP1216"
     primer_bind     4190..4214
                     /Sequence="CGATCGTTGTCAGAAGTAAGTTGGC"
                     /%GC="48.0"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1429"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:79n-e6a0gs8"">oSP1429</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:62c-e4f1i7m"">oSP1429</a>"
                     /label="oSP1429"
     primer_bind     complement(4226..4249)
                     /Sequence="GCAGTGCTGCCATAACCATGAGTG"
                     /%GC="54.2"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP257"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:9xu-dkzgvee"">oSP257</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1hg-cfz7esv"">oSP257</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6r9-dkzgu56"">oSP257</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2vu-cf0ayks"">oSP257</a>"
                     /label="oSP257"
     primer_bind     complement(4314..4339)
                     /Sequence="GCCGCATACACTATTCTCAGAATGAC"
                     /%GC="46.2"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1430"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:79s-e6a0gs9"">oSP1430</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:62h-e4f1i7n"">oSP1430</a>"
                     /label="oSP1430"
     primer_bind     4373..4392
                     /note="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     primer_bind     4373..4392
                     /Sequence="ATAATACCGCGCCACATAGC"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0005"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1ap-c6uht4m"">QB0005</a>"
                     /label="QB0005"
     primer_bind     4575..4598
                     /Sequence="GGGAATAAGGGCGACACGGAAATG"
                     /%GC="54.2"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP497"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:avq-dkzgvgj"">oSP497</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:7p5-dkzgu8r"">oSP497</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:9ie-cj5pty5"">oSP497</a>"
                     /label="oSP497"
     promoter        complement(4611..4715)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     4634..4659
                     /Sequence="GAAGCATTTATCAGGGTTATTGTCTC"
                     /%GC="38.5"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP864"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:c8m-dkzgvji"">oSP864</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:2qr-dck6bx5"">oSP864</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:921-dkzgucz"">oSP864</a>"
                     /label="oSP864"
     primer_bind     4665..4691
                     /Sequence="CGGATACATATTTGAATGTATTTAGAA"
                     /%GC="25.9"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP302"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:6y2-dkzgu5w"">oSP302</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:32n-cf0ayl5"">oSP302</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:a4n-dkzgvet"">oSP302</a>"
                     /label="oSP302"
     primer_bind     4718..4741
                     /Sequence="CATTTCCCCGAAAAGTGCCACCTG"
                     /%GC="54.2"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP99"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1x7-c6uhy1r"">oSP99</a>"
                     /label="oSP99"
     primer_bind     4724..4741
                     /Sequence="CCCGAAAAGTGCCACCTG"
                     /%GC="61.1"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0090"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1mn-c6uht6m"">QB0090</a>"
                     /label="QB0090"
     primer_bind     complement(4770..4793)
                     /Sequence="CTGAGAGTAGACCCTAGGGGATCG"
                     /%GC="58.3"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP735"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:bqp-dkzgvig"">oSP735</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:8k4-dkzgubj"">oSP735</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:88-d5zcyne"">oSP735</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:4et-d6qqpgr"">oSP735</a>"
                     /label="oSP735"
     primer_bind     complement(4793..4816)
                     /Sequence="ATGCGGCATCAGAGCAGATTGTAC"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1021"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:cuf-dkzgvky"">oSP1021</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:4ot-djp1nsx"">oSP1021</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:9nu-dkzguel"">oSP1021</a>"
                     /label="oSP1021"
     primer_bind     complement(4794..4813)
                     /note="pRS vectors, use to sequence yeast
                     selectable<br>marker"
                     /label="pRS-marker"
     primer_bind     complement(4794..4813)
                     /Sequence="CGGCATCAGAGCAGATTGTA"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="QB0092"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:1mx-c6uht6n"">QB0092</a>"
                     /label="QB0092"
     primer_bind     4817..4840
                     /Sequence="AGTTAAGCCAGTATCTGCTCCCTG"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2022.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="oSP1025"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:9oe-dkzguem"">oSP1025</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:4pd-djp1nt1"">oSP1025</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:cuz-dkzgvkz"">oSP1025</a>"
                     /label="oSP1025"
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 agatctcgag ctcggtacct aatacgacac actataagga aataagagag aaaagaagag
      661 taagaagaaa tataagagcc accatgagag ccctggtagt catccgcctg tcccgcgtca
      721 ccgatgctac gacttcaccg gagcgtcagc tggagtcttg ccagcagctc tgcgcccagc
      781 gcggctggga cgtcgtcggg gtagcggagg atctggacgt ctccggggcg gtcgatccgt
      841 tcgaccggaa gcgcagaccg aacctggccc ggtggctagc gttcgaggag caaccgttcg
      901 acgccatcgt ggcgtaccgg gtagaccggt tgacccgatc gatccggcat ctgcagcagc
      961 tggtccactg ggccgaggac cacaagaagc tggtcgtctc cgcgaccgaa gcgcacttcg
     1021 atacgacgac gccgtttgcg gcggtcgtca tcgcgcttat gggaacggtg gcgcagatgg
     1081 aattagaagc gatcaaagag cggaaccgtt cggctgcgca tttcaatatc cgcgccggga
     1141 aataccgagg atccctgccg ccgtggggat acctgcctac gcgcgtggac ggggagtggc
     1201 ggctggtgcc ggaccctgtg cagcgagagc gcatcctcga ggtgtatcac cgcgtcgtcg
     1261 acaaccacga gccgctgcac ctggtggccc acgacctgaa ccggcgtggt gtcctgtcgc
     1321 cgaaggacta cttcgcgcag ctgcaaggcc gcgagccgca gggccggaag tggtcggcta
     1381 ccgcgctgaa gcgatcgatg atctccgagg cgatgctcgg gtacgcgact ctgaacggta
     1441 agaccgtccg agacgacgac ggagccccgc tggtgcgggc tgagccgatc ctgacccgtg
     1501 agcagctgga ggcgctgcgc gccgagctcg tgaagacctc ccgggcgaag cccgcggtgt
     1561 ctaccccgtc gctgctgctg cgggtgttgt tctgcgcggt gtgcggggag cccgcgtaca
     1621 agttcgccgg gggaggacgt aagcacccgc gctaccgctg ccgctcgatg gggttcccga
     1681 agcactgcgg gaacggcacg gtggcgatgg ccgagtggga cgcgttctgc gaggagcagg
     1741 tgctggatct gctcggggac gcggagcgtc tggagaaagt ctgggtagcc ggctcggact
     1801 ccgcgatcga actcgcggag gtgaacgcgg agctggtgga cctgacgtcg ctgatcggct
     1861 ccccggccta ccgggccggc tctccgcagc gagaagcact ggatgcccgt attgcggcgc
     1921 tggccgcgcg gcaagaggag ctggagggtc tagaggctcg cccgtctggc tgggagtggc
     1981 gcgagaccgg gcagcggttc ggggactggt ggcgggagca ggacaccgcg gcaaagaaca
     2041 cctggcttcg gtcgatgaac gttcggctga cgttcgacgt ccgcggcggg ctgactcgca
     2101 cgatcgactt cggggatctg caggagtacg agcagcatct caggctcggc agcgtggtcg
     2161 aacggctaca caccgggatg tcgtagtaat taattaagct gccttctgcg gggcttgcct
     2221 tctggccatg cccttcttct ctcccttgca cctgtacctc ttggtctttg aataaagcct
     2281 gagtaggaag cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2341 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgagaaaatt
     2401 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2461 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2521 tctgaggcgg aaagaaccag ctggggctcg ataccgtcga cctctagcta gagcttggcg
     2581 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2641 atacgagccg gaagcataaa gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca
     2701 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2761 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2821 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2881 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2941 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3001 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3061 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3121 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3181 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3241 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3301 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3361 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3421 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3481 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3541 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3601 acggggtctg acactcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3661 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3721 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3781 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3841 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3901 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3961 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4021 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4081 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4141 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4201 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4261 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4321 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4381 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4441 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4501 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4561 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4621 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4681 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4741 gacgtcgacg gatcgggaga tcgatctccc gatcccctag ggtctactct cagtacaatc
     4801 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     4861 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     4921 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     4981 cgtt
//
