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
//
