LOCUS       Exported                5755 bp DNA     circular SYN 10-SEP-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5755)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    69..97
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    105..121
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             191..1366
                     /codon_start=1
                     /gene="malE"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This gene encodes a signal sequence that can direct 
                     MBP to the bacterial periplasm."
                     /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
                     LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
                     ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
                     ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
                     DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
                     KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKD
                     PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             1415..1429
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     terminator      1504..1590
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1682..1709
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1737..1827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1828..2688
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2776..3364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3734..3925)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        4421..4498
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4499..5581
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
       61 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca
      121 atttcacaca ggaaacagcc agtccgttta ggtgttttca cgagcaattg accaacaagg
      181 accatagatt atgaaaataa aaacaggtgc acgcatcctc gcattatccg cattaacgac
      241 gatgatgttt tccgcctcgg ctctcgccaa aatcgaagaa ggtaaactgg taatctggat
      301 taacggcgat aaaggctata acggtctcgc tgaagtcggt aagaaattcg agaaagatac
      361 cggaattaaa gtcaccgttg agcatccgga taaactggaa gagaaattcc cacaggttgc
      421 ggcaactggc gatggccctg acattatctt ctgggcacac gaccgctttg gtggctacgc
      481 tcaatctggc ctgttggctg aaatcacccc ggacaaagcg ttccaggaca agctgtatcc
      541 gtttacctgg gatgccgtac gttacaacgg caagctgatt gcttacccga tcgctgttga
      601 agcgttatcg ctgatttata acaaagatct gctgccgaac ccgccaaaaa cctgggaaga
      661 gatcccggcg ctggataaag aactgaaagc gaaaggtaag agcgcgctga tgttcaacct
      721 gcaagaaccg tacttcacct ggccgctgat tgctgctgac gggggttatg cgttcaagta
      781 tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat aacgctggcg cgaaagcggg
      841 tctgaccttc ctggttgacc tgattaaaaa caaacacatg aatgcagaca ccgattactc
      901 catcgcagaa gctgccttta ataaaggcga aacagcgatg accatcaacg gcccgtgggc
      961 atggtccaac atcgacacca gcaaagtgaa ttatggtgta acggtactgc cgaccttcaa
     1021 gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg ccgccagtcc
     1081 gaacaaagag ctggcaaaag agttcctcga aaactatctg ctgactgatg aaggtctgga
     1141 agcggttaat aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg aggaagagtt
     1201 ggtgaaagat ccgcgtattg ccgccactat ggaaaacgcc cagaaaggtg aaatcatgcc
     1261 gaacatcccg cagatgtccg ctttctggta tgccgtgcgt actgcggtga tcaacgccgc
     1321 cagcggtcgt cagactgtcg atgaagccct gaaagacgcg cagactaatt cgagctcgaa
     1381 caacaacaac aataacaata acaacaacct cggggatgac gatgacaagg taccgcatat
     1441 gtccatgggc ggccgcgata tcgtcgacgg atccgaattc cctgcaggta attaaataag
     1501 cttcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     1561 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     1621 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     1681 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttc ggtccgttgt
     1741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1801 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1861 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1921 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1981 ggtaagatcc ttgagagttt tcgccccgaa gaacgtttcc caatgatgag cacttttaaa
     2041 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     2101 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2161 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2221 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2281 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2341 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     2401 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2461 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2521 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2581 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2641 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2701 gtttactcat atatacttta gattgatttc cttaggactg agcgtcaacc ccgtagaaaa
     2761 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2821 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2881 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2941 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3001 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3061 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3421 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3481 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3541 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3601 ataaggtgca ctgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     3661 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3721 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     3781 aaagctcatc agcgtggtcg tgcagcgatt cacagatgtc tgcctgttca tccgcgtcca
     3841 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     3901 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     3961 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     4021 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     4081 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     4141 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     4201 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     4261 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     4321 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     4381 tgcgcacccg tggccaggac ccaacgctgc ccgaaattcc gacaccatcg aatggtgcaa
     4441 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
     4501 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     4561 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     4621 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     4681 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     4741 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     4801 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     4861 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     4921 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     4981 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     5041 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     5101 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     5161 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     5221 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     5281 gctgcgcgtt ggtgcggata tttcggtagt gggatacgac gataccgaag acagctcatg
     5341 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     5401 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     5461 ctcactggtg aaaagaaaaa ccaccctggc gcccaataca caaaccgcct ctccccgcgc
     5521 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5581 agcgcaacgc aattaatgta agttagctca ctcattaggc acaattctca tgtttgacag
     5641 cttatcatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
     5701 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcac
//
