LOCUS       Exported                5680 bp DNA     circular SYN 11-SEP-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5680)
  AUTHORS   11111111
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5680)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 11, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(4356..5680,1..4355)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    81..109
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    117..133
                     /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             203..1303
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             1352..1366
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     terminator      1441..1527
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1619..1646
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1674..1764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1765..2625
                     /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      2713..3301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3671..3862)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        4358..4435
                     /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             4436..5518
                     /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 ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc
       61 aaatattctg aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt
      121 gagcggataa caatttcaca caggaaacag ccagtccgtt taggtgtttt cacgagcaat
      181 tgaccaacaa ggaccataga ttatgaaaat cgaagaaggt aaactggtaa tctggattaa
      241 cggcgataaa ggctataacg gtctcgctga agtcggtaag aaattcgaga aagataccgg
      301 aattaaagtc accgttgagc atccggataa actggaagag aaattcccac aggttgcggc
      361 aactggcgat ggccctgaca ttatcttctg ggcacacgac cgctttggtg gctacgctca
      421 atctggcctg ttggctgaaa tcaccccgga caaagcgttc caggacaagc tgtatccgtt
      481 tacctgggat gccgtacgtt acaacggcaa gctgattgct tacccgatcg ctgttgaagc
      541 gttatcgctg atttataaca aagatctgct gccgaacccg ccaaaaacct gggaagagat
      601 cccggcgctg gataaagaac tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca
      661 agaaccgtac ttcacctggc cgctgattgc tgctgacggg ggttatgcgt tcaagtatga
      721 aaacggcaag tacgacatta aagacgtggg cgtggataac gctggcgcga aagcgggtct
      781 gaccttcctg gttgacctga ttaaaaacaa acacatgaat gcagacaccg attactccat
      841 cgcagaagct gcctttaata aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg
      901 gtccaacatc gacaccagca aagtgaatta tggtgtaacg gtactgccga ccttcaaggg
      961 tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa
     1021 caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg actgatgaag gtctggaagc
     1081 ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag tcttacgagg aagagttggt
     1141 gaaagatccg cgtattgccg ccactatgga aaacgcccag aaaggtgaaa tcatgccgaa
     1201 catcccgcag atgtccgctt tctggtatgc cgtgcgtact gcggtgatca acgccgccag
     1261 cggtcgtcag actgtcgatg aagccctgaa agacgcgcag actaattcga gctcgaacaa
     1321 caacaacaat aacaataaca acaacctcgg ggatgacgat gacaaggtac cgcatatgtc
     1381 catgggcggc cgcgatatcg tcgacggatc cgaattccct gcaggtaatt aaataagctt
     1441 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     1501 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     1561 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
     1621 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctctttcggt ccgttgttta
     1681 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1741 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1801 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1861 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1921 aagatccttg agagttttcg ccccgaagaa cgtttcccaa tgatgagcac ttttaaagtt
     1981 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     2041 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2101 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2161 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2221 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2281 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2341 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2401 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2461 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2521 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2581 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2641 tactcatata tactttagat tgatttcctt aggactgagc gtcaaccccg tagaaaagat
     2701 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2761 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2821 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2881 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2941 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3001 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3061 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3121 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3181 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3241 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3301 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3361 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3421 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3481 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatata
     3541 aggtgcactg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3601 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3661 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3721 gctcatcagc gtggtcgtgc agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3781 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3841 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     3901 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     3961 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     4021 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     4081 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     4141 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     4201 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     4261 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     4321 gcacccgtgg ccaggaccca acgctgcccg aaattccgac accatcgaat ggtgcaaaac
     4381 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     4441 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     4501 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     4561 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     4621 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     4681 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     4741 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     4801 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     4861 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     4921 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     4981 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     5041 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     5101 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     5161 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     5221 gcgcgttggt gcggatattt cggtagtggg atacgacgat accgaagaca gctcatgtta
     5281 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     5341 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     5401 actggtgaaa agaaaaacca ccctggcgcc caatacacaa accgcctctc cccgcgcgtt
     5461 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5521 gcaacgcaat taatgtaagt tagctcactc attaggcaca attctcatgt ttgacagctt
     5581 atcatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt
     5641 atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg
//
