LOCUS       Exported                8717 bp DNA     circular SYN 16-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8717)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8717)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2160..2177
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2394..2480
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2572..2599
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2618..2709
                     /label=AmpR promoter
     rep_origin      complement(3096..3684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6528..7184)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(8337..8414)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        8644..8672
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    8680..8696
                     /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)."
ORIGIN
        1 aattaagctt aaaggaggac aaccatgcca ctgttcggat ccttctccga ccgcgtcaac
       61 cgtgcagtgc tctacaggat ctgtgcaaag gaggacaact aatgtttaac aaccgtatcc
      121 gcactgcagc tcttgctggt gcaatcgcaa tctccaccgc agcttccggc gttgctatcc
      181 cagcattcgc tctcccggcg aaccaccgat cggggtcgat cgaggacgtc gagcacatcg
      241 tcgtcctgat gcaggagaac cgttccttcg accactactt cggcacactg cgcggggtcc
      301 gcggcttcgg cgacccgcat ccggtgcgac tggacgacgg caggtcggtg tggcaccagc
      361 gcaagggcga cggcaccgag gtactaccct tccacccgga ggccgacgac ctcggtatgc
      421 agttcctcga agggctcccg cacggctggt ccgacggcca ggacgcctac cacgacgcca
      481 agtacgaccg ctggctgccc gccaagggca ccaccaccat ggcgtacctg acccgcgagg
      541 acatcccctt ccactacgcg ctcgccgaca ccttcaccgt ctgcgacgcc taccactgct
      601 ccttcatcgg ctccaccgac cccaaccgct actacctgtg gtcggggcac acgggcaacg
      661 acggcgcggg cggcggcccg gtcctcggca acgacgaact cggctacgac tggacgacgt
      721 accccgagcg cctggaggcg gccggggtct cctggaagat ctaccaggac gtcggcgacg
      781 gcctcgacgc ggccggccac tggggctgga tcgacgacgc ctaccggggc aactacggcg
      841 acaactccct gctgtacttc aacaagtacc ggaacgcgaa gcccggcgac ccgctgtacg
      901 acaaggcccg caccggcacc gacgcgaagc gcggcgaggg ctacttcgac cggctccgtg
      961 ccgacgtcaa ggcgggcaag ctcccggaga tctcctggat agccgccccg gaggcattct
     1021 ccgagcactc caactggccg tcgaactacg gcgcctggta catctcccag gtcctggacg
     1081 cgctcacctc caacccgaag gtgtgggcga agaccgccct gttcatcacg tacgacgaga
     1141 acgacggctt cttcgaccac gtggtgccgc cgctgccgcc gaagtccgcc gcgcagggcc
     1201 ggtccaccgt cgacgtctcc ctggacgtct tcgagggcag cgccaagcac cgcgagggct
     1261 tctacgggct cggcccgcgc gtgcccatgc tggtcgtctc gccgtggagc aagggcggct
     1321 tcgtctgctc cgagaccttc gaccacacct cggtcatccg cttcatggaa cgccgcttcg
     1381 gcgtccacga gccgaacatc tcaccctggc ggcgcgccgt ctgcggcgac ctgacctccg
     1441 ccttcgactt ctcccgcacg gacagccgcc cggcccggct gccggacacc gacgcctacg
     1501 agccgccgga ccgcgaacgc caccccgact accgtccgac gccgcccgcg gaccccggca
     1561 tgccccggca ggaacgcggg acacgcccca cccggccgct gcgctacgcc ccgcacgtcg
     1621 acggcgccgt ggacacggcg gccgggaagt tcacgctggc cttcgcctcc gggccccggg
     1681 ccggtgccgc cttcctcgtc acctccggca accgcaccga cggcccctgg acgtacacca
     1741 ccgaggccgg caaggacatc gcggacacct ggaactcggc ctactccggc ggctcctacg
     1801 acctgaccgt gcacggcccg gccggcttcc tgcgggtctt ccggggtgcc aacgaggtcg
     1861 ccggacccgc cgggcccgag gtcaccgcac ggcaccgcgg cgacgacctc cggctgaccc
     1921 tcacccacac cgggaccggc accgtccggc tccggatcgc gaacgcccac gacggccgca
     1981 cccggtccgt caccgtccgc cccggcgcca ccgtgcacca caccgtcggc ctcgcgcgca
     2041 gccggcgctg gtacgacgtg acggtcacct ccgagaccgc cccggccttc ctgcgccgct
     2101 tcgccgggca cgtggagaac ggccggccgg ggaccagcga cccggcgatc gtcacggacc
     2161 atcatcacca tcaccattga gaattcagct tggctgtttt ggcggatgag agaagatttt
     2221 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     2281 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     2341 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     2401 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2461 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     2521 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     2581 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
     2641 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2701 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2761 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2821 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2881 tttcgccccg aagaacgttt tccaatgatg agcacttttg cttcctcgct cactgactcg
     2941 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3001 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3061 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3121 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3181 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3241 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3301 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3361 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3421 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3481 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3541 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3601 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3661 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3721 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3781 acctagatcc ttttggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     3841 atccagccat tcggggtcgt tcactggttc ccctttctga tttctggcat agaagaaccc
     3901 ccgtgaactg tgtggttccg ggggttgctg atttttgcga gacttctcgc gcaattccct
     3961 agcttaggtg aaaacaccat gaaacactag ggaaacaccc atgaaacacc cattagggca
     4021 gtagggcggc ttcttcgtct agggcttgca tttgggcggt gatctggtct ttagcgtgtg
     4081 aaagtgtgtc gtaggtggcg tgctcaatgc actcgaacgt cacgtcattt accgggtcac
     4141 ggtgggcaaa gagaactagt gggttagaca ttgttttcct cgttgtcggt ggtggtgagc
     4201 ttttctagcc gctcggtaaa cgcggcgatc atgaactctt ggaggttttc accgttctgc
     4261 atgcctgcgc gcttcatgtc ctcacgtagt gccaaaggaa cgcgtgcggt gaccacgacg
     4321 ggcttagcct ttgcctgcgc ttctagtgct tcgatggtgg cttgtgcctg cgcttgctgc
     4381 gcctgtagtg cctgttgagc ttcttgtagt tgctgttcta gctgtgcctt ggttgccatg
     4441 ctttaagact ctagtagctt tcctgcgata tgtcatgcgc atgcgtagca aacattgtcc
     4501 tgcaactcat tcattatgtg cagtgctcct gttactagtc gtacatactc atatttacct
     4561 agtctgcatg cagtgcatgc acatgcagtc atgtcgtgct aatgtgtaaa acatgtacat
     4621 gcagattgct gggggtgcag ggggcggagc caccctgtcc atgcggggtg tggggcttgc
     4681 cccgccggta cagacagtga gcaccggggc acctagtcgc ggataccccc cctaggtatc
     4741 ggacacgtaa ccctcccatg tcgatgcaaa tctttaacat tgagtacggg taagctggca
     4801 cgcatagcca agctaggcgg ccaccaaaca ccactaaaaa ttaatagtcc ctagacaaga
     4861 caaacccccg tgcgagctac caactcatat gcacgggggc cacataaccc gaaggggttt
     4921 caattgacaa ccatagcact agctaagaca acgggcacaa cacccgcaca aactcgcact
     4981 gcgcaacccc gcacaacatc gggtctaggt aacactgaaa tagaagtgaa cacctctaag
     5041 gaaccgcagg tcaatgaggg ttctaaggtc actcgcgcta gggcgtggcg taggcaaaac
     5101 gtcatgtaca agatcaccaa tagtaaggct ctggcggggt gccataggtg gcgcagggac
     5161 gaagctgttg cggtgtcctg gtcgtctaac ggtgcttcgc agtttgaggg tctgcaaaac
     5221 tctcactctc gctgggggtc acctctggct gaattggaag tcatgggcga acgccgcatt
     5281 gagctggcta ttgctactaa gaatcacttg gcggcgggtg gcgcgctcat gatgtttgtg
     5341 ggcactgttc gacacaaccg ctcacagtca tttgcgcagg ttgaagcggg tattaagact
     5401 gcgtactctt cgatggtgaa aacatctcag tggaagaaag aacgtgcacg gtacggggtg
     5461 gagcacacct atagtgacta tgaggtcaca gactcttggg cgaacggttg gcacttgcac
     5521 cgcaacatgc tgttgttctt ggatcgtcca ctgtctgacg atgaactcaa ggcgtttgag
     5581 gattccatgt tttcccgctg gtctgctggt gtggttaagg ccggtatgga cgcgccactg
     5641 cgtgagcacg gggtcaaact tgatcaggtg tctacctggg gtggagacgc tgcgaaaatg
     5701 gcaacctacc tcgctaaggg catgtctcag gaactgactg gctccgctac taaaaccgcg
     5761 tctaaggggt cgtacacgcc gtttcagatg ttggatatgt tggccgatca aagcgacgcc
     5821 ggcgaggata tggacgctgt tttggtggct cggtggcgtg agtatgaggt tggttctaaa
     5881 aacctgcgtt cgtcctggtc acgtggggct aagcgtgctt tgggcattga ttacatagac
     5941 gctgatgtac gtcgtgaaat ggaagaagaa ctgtacaagc tcgccggtct ggaagcaccg
     6001 gaacgggtcg aatcaacccg cgttgctgtt gctttggtga agcccgatga ttggaaactg
     6061 attcagtctg atttcgcggt taggcagtac gttctcgatt gcgtggataa ggctaaggac
     6121 gtggccgctg cgcaacgtgt cgctaatgag gtgctggcaa gtctgggtgt ggattccacc
     6181 ccgtgcatga tcgttatgga tgatgtggac ttggacgcgg ttctgcctac tcatggggac
     6241 gctactaagc gtgatctgaa tgcggcggtg ttcgcgggta atgagcagac tattcttcgc
     6301 acccactaaa agcggcataa accccgttcg atattttgtg cgatgaattt atggtcaatg
     6361 tcgcgggggc aaactatgat gggtcttgtt gttggcgtcc cggaaaacga ttccgaagcc
     6421 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
     6481 ttggtcggtc atttcgaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
     6541 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     6601 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     6661 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     6721 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     6781 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     6841 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     6901 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     6961 atacgtaact ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     7021 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     7081 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     7141 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     7201 cctgaaaatc tcgtcgaagc tcggcggatt tgtcctactc aagctgatcc gacaaaatcc
     7261 acacattatc ccaggtgtcc ggatcggtca aatacgctgc cagctcatag accgtatcca
     7321 aagcatccgg ggctgatccc cggcgccagg gtggtttttc ttttcaccag tgagacgggc
     7381 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     7441 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     7501 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     7561 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     7621 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     7681 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     7741 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     7801 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     7861 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     7921 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     7981 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     8041 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     8101 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     8161 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     8221 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     8281 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     8341 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac
     8401 cattcgatgg tgtcaacgta aatgccgctt cgccttcgcg cgcgaattgc aagctgatcc
     8461 gggcttatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
     8521 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg
     8581 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag
     8641 ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc
     8701 acacaggaaa cagaatt
//
