LOCUS       Exported                5489 bp ds-DNA     circular SYN 13-SEP-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5489)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5489)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 13, 2025 from SnapGene 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5489
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          join(461..5489,1..460)
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        84..102
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    103..127
                     /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)."
     RBS             142..164
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     172..237
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /note="/codon_start=1"
                     /note="/translation=MKYLLPTAAAGLLLLAAQPAMA"
     CDS             304..321
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      388..435
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      472..927
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        953..1057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1058..1918
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2089..2677
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2863..3002
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3104..3295)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4104..5186)
                     /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"
     promoter        complement(5187..5264)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
       61 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
      121 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgaaatac
      181 ctgctgccga ccgctgctgc tggtctgctg ctcctcgctg cccagccggc gatggccatg
      241 gatatcggaa ttaattcgga tccgaattcg agctccgtcg acaagcttgc ggccgcactc
      301 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag
      361 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
      421 ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct
      481 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
      541 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
      601 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
      661 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
      721 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
      781 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
      841 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
      901 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
      961 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1021 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1081 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1141 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1201 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1261 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     1321 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1381 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1441 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1501 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1561 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     1621 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1681 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1741 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     1801 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1861 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1921 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1981 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2041 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2101 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2161 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2221 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2281 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2341 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2401 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2461 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2521 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2581 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2641 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2701 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2761 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2821 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     2881 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     2941 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3001 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3061 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3121 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3181 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3241 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     3301 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     3361 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     3421 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     3481 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     3541 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     3601 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     3661 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     3721 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga
     3781 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     3841 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     3901 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     3961 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4021 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     4081 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4141 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     4201 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     4261 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     4321 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4381 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4441 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4501 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4561 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4621 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4681 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4741 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4801 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     4861 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     4921 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     4981 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     5041 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     5101 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     5161 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     5221 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     5281 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5341 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     5401 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5461 ttccccatcg gtgatgtcgg cgatatagg
//
