LOCUS       Exported                6645 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 6645)
  AUTHORS   11111111
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    97..125
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    133..149
                     /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             219..1319
                     /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             1368..1379
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1427..1443)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1792..1878
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1970..1997
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2016..2107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2108..2968
                     /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      complement(3068..3523)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      3634..4222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4408..4550
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4652..4843)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        5339..5416
                     /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             5417..6499
                     /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 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
       61 tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca tcggctcgta
      121 taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagccag tccgtttagg
      181 tgttttcacg agcacttcac caacaaggac catagcatat gaaaatcgaa gaaggtaaac
      241 tggtaatctg gattaacggc gataaaggct ataacggtct cgctgaagtc ggtaagaaat
      301 tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc ggataaactg gaagagaaat
      361 tcccacaggt tgcggcaact ggcgatggcc ctgacattat cttctgggca cacgaccgct
      421 ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg
      481 acaagctgta tccgtttacc tgggatgccg tacgttacaa cggcaagctg attgcttacc
      541 cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga tctgctgccg aacccgccaa
      601 aaacctggga agagatcccg gcgctggata aagaactgaa agcgaaaggt aagagcgcgc
      661 tgatgttcaa cctgcaagaa ccgtacttca cctggccgct gattgctgct gacgggggtt
      721 atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg gataacgctg
      781 gcgcgaaagc gggtctgacc ttcctggttg acctgattaa aaacaaacac atgaatgcag
      841 acaccgatta ctccatcgca gaagctgcct ttaataaagg cgaaacagcg atgaccatca
      901 acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt gaattatggt gtaacggtac
      961 tgccgacctt caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta
     1021 acgccgccag tccgaacaaa gagctggcaa aagagttcct cgaaaactat ctgctgactg
     1081 atgaaggtct ggaagcggtt aataaagaca aaccgctggg tgccgtagcg ctgaagtctt
     1141 acgaggaaga gttggtgaaa gatccgcgga ttgccgccac tatggaaaac gcccagaaag
     1201 gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg
     1261 tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc cctgaaagac gcgcagacta
     1321 attcgagctc gaacaacaac aacaataaca ataacaacaa cctcgggatc gagggaagga
     1381 tttcagaatt cggatcctct agagtcgacc tgcaggcaag cttggcactg gccgtcgttt
     1441 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     1501 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1561 tgcgcagcct gaatggcgaa tggcagcttg gctgttttgg cggatgagat aagattttca
     1621 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     1681 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     1741 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     1801 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1861 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     1921 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     1981 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
     2041 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2101 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2161 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2221 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2281 tcgccccgaa gaacgtttcc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2341 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2401 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2461 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2521 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2581 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2641 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2701 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2761 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2821 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2881 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2941 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3001 gattgattta ccccggttga taatcagaaa agccccaaaa acaggaagat tgtataagca
     3061 aatatttaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3121 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3181 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3241 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3301 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3361 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3421 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3481 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa
     3541 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3601 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3661 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3721 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3781 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3841 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3901 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3961 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4021 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4081 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4141 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4201 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4261 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4321 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4381 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4441 cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4501 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     4561 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     4621 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     4681 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg cagcgattca cagatgtctg
     4741 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     4801 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     4861 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     4921 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     4981 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
     5041 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
     5101 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     5161 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     5221 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     5281 acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gaaattccga
     5341 caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     5401 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     5461 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     5521 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     5581 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     5641 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     5701 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     5761 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     5821 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     5881 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     5941 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     6001 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     6061 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     6121 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     6181 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcagtagtgg gatacgacga
     6241 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     6301 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     6361 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     6421 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     6481 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     6541 aattctcatg tttgacagct tatcatcgac tgcacggtgc accaatgctt ctggcgtcag
     6601 gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcac
//
