LOCUS Exported 2687 bp ds-DNA circular SYN 24-SEP-2020 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2687) AUTHORS Keoni Gandall TITLE Direct Submission JOURNAL Exported Thursday, Sep 24, 2020 from SnapGene 5.0.7 https://www.snapgene.com FEATURES Location/Qualifiers source 1..2687 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 293..482 /label=spacer /note="Leftover from pUC19" misc_feature 297..316 /label=anneal_sequence CDS complement(483..1343) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" misc_feature 627 /label=C->A /note="removes BsaI site" promoter complement(1344..1448) /gene="bla" /label=AmpR promoter misc_feature 1449..1463 /label=AarI primer_bind 1464..1480 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" terminator 1481..1512 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" primer_bind complement(1510..1528) /label=SLiCE-rev misc_feature 1529..1548 /label=BbsI-BtgZI CDS 1874..2179 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" misc_feature 2203..2222 /label=BbsI-BtgZI primer_bind 2223..2241 /label=SLiCE-for primer_bind complement(2242..2260) /label=T7 Reverse /note="https://www.elimbio.com/elim's_primer.htm" terminator 2256..2303 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(2304..2324) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2308..2324) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" misc_feature 2325..2339 /label=AarI CDS 2340..2390 /codon_start=1 /label=spacer /translation="MTHINKFREESPEECH" rep_origin complement(join(2391..2687,1..292)) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 61 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaaggcct 301 actattagca acaacgatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 361 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 421 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 481 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 541 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 601 cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt atcagcaata 661 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 721 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 781 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 841 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 901 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 961 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 1021 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 1081 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 1141 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 1201 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 1261 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 1321 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 1381 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 1441 gttccgcgca cctgcaccag tcagtaaaac gacggccagt agtcaaaagc ctccgaccgg 1501 aggcttttga cttggttcag gtggagtggg agtagtcttc gccatcgcta ctaaaagcca 1561 gataacagta tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt 1621 atacccgaag tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga 1681 cagcgacagc tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc 1741 acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag 1801 gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac 1861 aggggctggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct 1921 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct 1981 ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat 2041 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat 2101 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct 2161 gatgttctgg ggaatataaa tgtcaggctc ccttatacac aggcgatgtt gaagaccacg 2221 ctgaggtgtc aatcgtcgga gccgctgagc aataactagc ataacccctt ggggcctcta 2281 aacgggtctt gaggggtttt ttgcatggtc atagctgttt cctgagagct tggcaggtga 2341 tgacacacat taacaaattt cgtgaggagt ctccagaaga atgccattaa tttccatagg 2401 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 2461 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 2521 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 2581 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 2641 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccgg //