LOCUS       Exported                6591 bp DNA     circular SYN 12-AUG-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6591)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6591)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6591)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(5..61)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      269..355
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      269..355
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      447..474
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      447..474
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        493..584
                     /label=AmpR promoter
     rep_origin      complement(971..1559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             2979..4184
                     /codon_start=1
                     /label=repBl1
                     /translation="MYKITNSKALAGCHRWRRDEAVAVSWSSNGASQFEGLQNSHSRWG
                     SPLAELEVMGERRIELAIATKNHLAAGGALMMFVGTVRHNRSQSFAQVEAGIKTAYSSM
                     VKTSQWKKERARYGVEHTYSDYEVTDSWANGWHLHRNMLLFLDRPLSDDELKAFEDSMF
                     SRWSAGVVKAGMDAPLREHGVKLDQVSTWGGDAAKMATYLAKGMSQELTGSATKTASKG
                     SYTPFQMLDMLADQSDAGEDMDAVLVARWREYEVGSKNLRSSWSRGAKRALGIDYIDAD
                     VRREMEEELYKLAGLEAPERVESTRVAVALVKPDDWKLIQSDFAVRQYVLDCVDKAKDV
                     AAAQRVANEVLASLGVDSTPCMIVMDDVDLDAVLPTHGDATKRDLNAAVFAGNEQTILR
                     TH"
     CDS             complement(4400..5059)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(5272..6087)
                     /codon_start=1
                     /label=LacI
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTTWTACCNSLRARR"
     promoter        complement(6211..6288)
                     /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        6518..6546
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    6554..6570
                     /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 gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg agctcgaatt
       61 cagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
      121 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
      181 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
      241 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
      301 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
      361 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
      421 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
      481 tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      541 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
      601 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
      661 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
      721 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
      781 tgatgagcac ttttgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      841 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      901 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      961 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1021 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1081 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1141 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1201 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1261 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1321 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1381 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1441 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1501 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1561 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1621 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg gggtgggcga
     1681 agaactccag catgagatcc ccgcgctgga ggatcatcca gccattcggg gtcgttcact
     1741 ggttcccctt tctgatttct ggcatagaag aacccccgtg aactgtgtgg ttccgggggt
     1801 tgctgatttt tgcgagactt ctcgcgcaat tccctagctt aggtgaaaac accatgaaac
     1861 actagggaaa cacccatgaa acacccatta gggcagtagg gcggcttctt cgtctagggc
     1921 ttgcatttgg gcggtgatct ggtctttagc gtgtgaaagt gtgtcgtagg tggcgtgctc
     1981 aatgcactcg aacgtcacgt catttaccgg gtcacggtgg gcaaagagaa ctagtgggtt
     2041 agacattgtt ttcctcgttg tcggtggtgg tgagcttttc tagccgctcg gtaaacgcgg
     2101 cgatcatgaa ctcttggagg ttttcaccgt tctgcatgcc tgcgcgcttc atgtcctcac
     2161 gtagtgccaa aggaacgcgt gcggtgacca cgacgggctt agcctttgcc tgcgcttcta
     2221 gtgcttcgat ggtggcttgt gcctgcgctt gctgcgcctg tagtgcctgt tgagcttctt
     2281 gtagttgctg ttctagctgt gccttggttg ccatgcttta agactctagt agctttcctg
     2341 cgatatgtca tgcgcatgcg tagcaaacat tgtcctgcaa ctcattcatt atgtgcagtg
     2401 ctcctgttac tagtcgtaca tactcatatt tacctagtct gcatgcagtg catgcacatg
     2461 cagtcatgtc gtgctaatgt gtaaaacatg tacatgcaga ttgctggggg tgcagggggc
     2521 ggagccaccc tgtccatgcg gggtgtgggg cttgccccgc cggtacagac agtgagcacc
     2581 ggggcaccta gtcgcggata ccccccctag gtatcggaca cgtaaccctc ccatgtcgat
     2641 gcaaatcttt aacattgagt acgggtaagc tggcacgcat agccaagcta ggcggccacc
     2701 aaacaccact aaaaattaat agtccctaga caagacaaac ccccgtgcga gctaccaact
     2761 catatgcacg ggggccacat aacccgaagg ggtttcaatt gacaaccata gcactagcta
     2821 agacaacggg cacaacaccc gcacaaactc gcactgcgca accccgcaca acatcgggtc
     2881 taggtaacac tgaaatagaa gtgaacacct ctaaggaacc gcaggtcaat gagggttcta
     2941 aggtcactcg cgctagggcg tggcgtaggc aaaacgtcat gtacaagatc accaatagta
     3001 aggctctggc ggggtgccat aggtggcgca gggacgaagc tgttgcggtg tcctggtcgt
     3061 ctaacggtgc ttcgcagttt gagggtctgc aaaactctca ctctcgctgg gggtcacctc
     3121 tggctgaatt ggaagtcatg ggcgaacgcc gcattgagct ggctattgct actaagaatc
     3181 acttggcggc gggtggcgcg ctcatgatgt ttgtgggcac tgttcgacac aaccgctcac
     3241 agtcatttgc gcaggttgaa gcgggtatta agactgcgta ctcttcgatg gtgaaaacat
     3301 ctcagtggaa gaaagaacgt gcacggtacg gggtggagca cacctatagt gactatgagg
     3361 tcacagactc ttgggcgaac ggttggcact tgcaccgcaa catgctgttg ttcttggatc
     3421 gtccactgtc tgacgatgaa ctcaaggcgt ttgaggattc catgttttcc cgctggtctg
     3481 ctggtgtggt taaggccggt atggacgcgc cactgcgtga gcacggggtc aaacttgatc
     3541 aggtgtctac ctggggtgga gacgctgcga aaatggcaac ctacctcgct aagggcatgt
     3601 ctcaggaact gactggctcc gctactaaaa ccgcgtctaa ggggtcgtac acgccgtttc
     3661 agatgttgga tatgttggcc gatcaaagcg acgccggcga ggatatggac gctgttttgg
     3721 tggctcggtg gcgtgagtat gaggttggtt ctaaaaacct gcgttcgtcc tggtcacgtg
     3781 gggctaagcg tgctttgggc attgattaca tagacgctga tgtacgtcgt gaaatggaag
     3841 aagaactgta caagctcgcc ggtctggaag caccggaacg ggtcgaatca acccgcgttg
     3901 ctgttgcttt ggtgaagccc gatgattgga aactgattca gtctgatttc gcggttaggc
     3961 agtacgttct cgattgcgtg gataaggcta aggacgtggc cgctgcgcaa cgtgtcgcta
     4021 atgaggtgct ggcaagtctg ggtgtggatt ccaccccgtg catgatcgtt atggatgatg
     4081 tggacttgga cgcggttctg cctactcatg gggacgctac taagcgtgat ctgaatgcgg
     4141 cggtgttcgc gggtaatgag cagactattc ttcgcaccca ctaaaagcgg cataaacccc
     4201 gttcgatatt ttgtgcgatg aatttatggt caatgtcgcg ggggcaaact atgatgggtc
     4261 ttgttgttgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     4321 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaagggcacc
     4381 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     4441 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     4501 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     4561 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     4621 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     4681 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     4741 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     4801 tatcccatat caccagctca ccgtctttca ttgccatacg gaactccgga tgagcattca
     4861 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     4921 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     4981 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     5041 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgtc gaagctcggc
     5101 ggatttgtcc tactcaagct gatccgacaa aatccacaca ttatcccagg tgtccggatc
     5161 ggtcaaatac gctgccagct catagaccgt atccaaagca tccggggctg atccccggcg
     5221 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     5281 ggccctgaga gagttgcagc aagcggtcca cgtggtttgc cccagcaggc gaaaatcctg
     5341 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     5401 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     5461 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     5521 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     5581 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     5641 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     5701 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     5761 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     5821 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     5881 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     5941 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     6001 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     6061 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     6121 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     6181 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     6241 gcgctatcat gccataccgc gaaaggtttt gcaccattcg atggtgtcaa cgtaaatgcc
     6301 gcttcgcctt cgcgcgcgaa ttgcaagctg atccgggctt atcgactgca cggtgcacca
     6361 atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca
     6421 ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac
     6481 atcataacgg ttctggcaaa tattctgaaa tgagctgttg acaattaatc atcggctcgt
     6541 ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagaat t
//
