LOCUS       Exported                5715 bp DNA     circular SYN 20-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5715)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5715)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5715)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     423..439
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(508..524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(532..548)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(556..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(601..622)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(910..1498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1702..2502
                     /codon_start=1
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     CDS             complement(2939..3307)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(3340..3449)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3687..3698)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     protein_bind    3769..3868
                     /label=phage phi-C31 attP
                     /note="attachment site of phage phi-C31"
     CDS             3878..5692
                     /codon_start=1
                     /label=phage phi-C31 integrase
                     /note="integrase from phage phi-C31"
                     /translation="VDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
ORIGIN
        1 atctacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
       61 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
      121 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
      181 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattta tgcggtgtga
      241 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
      301 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      361 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      421 ttgtaaaacg acggccagtg ccaagcttgg gctgcaggtc gactctagag gatccgcggc
      481 cgcgcgcgat atcgaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
      541 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      601 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      661 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      721 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      781 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      841 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      901 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      961 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1021 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1081 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1141 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1201 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1261 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1321 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1381 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1441 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1501 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1561 gattttggtc atgagattat caaaaaggat cttcacctag atccttttgg ttcatgtgca
     1621 gctccatcag caaaagggga tgataagttt atcaccaccg actatttgca acagtgccgt
     1681 tgatcgtgct atgatcgact gatgtcatca gcggtggagt gcaatgtcgt gcaatacgaa
     1741 tggcgaaaag ccgagctcat cggtcagctt ctcaaccttg gggttacccc cggcggtgtg
     1801 ctgctggtcc acagctcctt ccgtagcgtc cggcccctcg aagatgggcc acttggactg
     1861 atcgaggccc tgcgtgctgc gctgggtccg ggagggacgc tcgtcatgcc ctcgtggtca
     1921 ggtctggacg acgagccgtt cgatcctgcc acgtcgcccg ttacaccgga ccttggagtt
     1981 gtctctgaca cattctggcg cctgccaaat gtaaagcgca gcgcccatcc atttgccttt
     2041 gcggcagcgg ggccacaggc agagcagatc atctctgatc cattgcccct gccacctcac
     2101 tcgcctgcaa gcccggtcgc ccgtgtccat gaactcgatg ggcaggtact tctcctcggc
     2161 gtgggacacg atgccaacac gacgctgcat cttgccgagt tgatggcaaa ggttccctat
     2221 ggggtgccga gacactgcac cattcttcag gatggcaagt tggtacgcgt cgattatctc
     2281 gagaatgacc actgctgtga gcgctttgcc ttggcggaca ggtggctcaa ggagaagagc
     2341 cttcagaagg aaggtccagt cggtcatgcc tttgctcggt tgatccgctc ccgcgacatt
     2401 gtggcgacag ccctgggtca actgggccga gatccgttga tcttcctgca tccgccagag
     2461 gcgggatgcg aagaatgcga tgccgctcgc cagtcgattg gctgagctca tgagcggaga
     2521 acgagatgac gttggagggg caaggtcgcg ctgattgctg gggcaacacg tggagcggat
     2581 cggggattgt ctttcttcag ctcgctgatg atatgctgac gctcaatgcc gtttggcctc
     2641 cgactaacga aaatcccgca tttggacggc tgatccgatt ggcacggcgg acggcgaatg
     2701 gcggagcaga cgctcgtccg ggggcaatga gatatgaaaa agcctgaact caccgcgacg
     2761 tatcgggccc tggccagcta gctagagtcg acctgcaggt ccccggggat cggtcttgcc
     2821 ttgctcgtcg gtgatgtact tcaccagctc cgcgaagtcg ctcttcttga tggagcgcat
     2881 ggggacgtgc ttggcaatca cgcgcacccc ccggccgttt tagcggctaa aaaagtcatg
     2941 gctctgccct cgggcggacc acgcccatca tgaccttgcc aagctcgtcc tgcttctctt
     3001 cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg cgcgggtcgt
     3061 cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg atgcgggcca
     3121 gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg ccgacggcca
     3181 gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc gctcttcgtt
     3241 cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc ttggtttcat
     3301 cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag
     3361 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
     3421 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
     3481 ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa
     3541 aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagatccgt cgacctgcag
     3601 gcatgcaagc tctagcgatt ccagacgtcc cgaaggcgtg gcgcggcttc cccgtgccgg
     3661 agcaatcgcc ctgggtgggt tacacgacgc ccctctatgg cccgtactga cggacacacc
     3721 gaagccccgg cggcaaccct cagcggatgc cccggggctt cacgttttcc caggtcagaa
     3781 gcggttttcg ggagtagtgc cccaactggg gtaacctttg agttctctca gttgggggcg
     3841 tagggtcgcc gacatgacac aaggggttgt gaccggggtg gacacgtacg cgggtgctta
     3901 cgaccgtcag tcgcgcgagc gcgaaaattc gagcgcagca agcccagcga cacagcgtag
     3961 cgccaacgaa gacaaggcgg ccgaccttca gcgcgaagtc gagcgcgacg ggggccggtt
     4021 caggttcgtc gggcatttca gcgaagcgcc gggcacgtcg gcgttcggga cggcggagcg
     4081 cccggagttc gaacgcatcc tgaacgaatg ccgcgccggg cggctcaaca tgatcattgt
     4141 ctatgacgtg tcgcgcttct cgcgcctgaa ggtcatggac gcgattccga ttgtctcgga
     4201 attgctcgcc ctgggcgtga cgattgtttc cactcaggaa ggcgtcttcc ggcagggaaa
     4261 cgtcatggac ctgattcacc tgattatgcg gctcgacgcg tcgcacaaag aatcttcgct
     4321 gaagtcggcg aagattctcg acacgaagaa ccttcagcgc gaattgggcg ggtacgtcgg
     4381 cgggaaggcg ccttacggct tcgagcttgt ttcggagacg aaggagatca cgcgcaacgg
     4441 ccgaatggtc aatgtcgtca tcaacaagct tgcgcactcg accactcccc ttaccggacc
     4501 cttcgagttc gagcccgacg taatccggtg gtggtggcgt gagatcaaga cgcacaaaca
     4561 ccttcccttc aagccgggca gtcaagccgc cattcacccg ggcagcatca cggggctttg
     4621 taagcgcatg gacgctgacg ccgtgccgac ccggggcgag acgattggga agaagaccgc
     4681 ttcaagcgcc tgggacccgg caaccgttat gcgaatcctt cgggacccgc gtattgcggg
     4741 cttcgccgct gaggtgatct acaagaagaa gccggacggc acgccgacca cgaagattga
     4801 gggttaccgc attcagcgcg acccgatcac gctccggccg gtcgagcttg attgcggacc
     4861 gatcatcgag cccgctgagt ggtatgagct tcaggcgtgg ttggacggca gggggcgcgg
     4921 caaggggctt tcccgggggc aagccattct gtccgccatg gacaagctgt actgcgagtg
     4981 tggcgccgtc atgacttcga agcgcgggga agaatcgatc aaggactctt accgctgccg
     5041 tcgccggaag gtggtcgacc cgtccgcacc tgggcagcac gaaggcacgt gcaacgtcag
     5101 catggcggca ctcgacaagt tcgttgcgga acgcatcttc aacaagatca ggcacgccga
     5161 aggcgacgaa gagacgttgg cgcttctgtg ggaagccgcc cgacgcttcg gcaagctcac
     5221 tgaggcgcct gagaagagcg gcgaacgggc gaaccttgtt gcggagcgcg ccgacgccct
     5281 gaacgccctt gaagagctgt acgaagaccg cgcggcaggc gcgtacgacg gacccgttgg
     5341 caggaagcac ttccggaagc aacaggcagc gctgacgctc cggcagcaag gggcggaaga
     5401 gcggcttgcc gaacttgaag ccgccgaagc cccgaagctt ccccttgacc aatggttccc
     5461 cgaagacgcc gacgctgacc cgaccggccc taagtcgtgg tgggggcgcg cgtcagtaga
     5521 cgacaagcgc gtgttcgtcg ggctcttcgt agacaagatc gttgtcacga agtcgactac
     5581 gggcaggggg cagggaacgc ccatcgagaa gcgcgcttcg atcacgtggg cgaagccgcc
     5641 gaccgacgac gacgaagacg acgcccagga cggcacggaa gacgtagcgg cgtagcgaga
     5701 cacccgggaa gcctg
//
