LOCUS       sequence-54724-e        3902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3902)
  TITLE     OPPF-UK pOPIN vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3902)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pOPINCDF
FEATURES             Location/Qualifiers
     source          1..3902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             77..94
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    168..189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(204..209,210..227,228..234)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    242..258
                     /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)."
     primer_bind     247..269
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     266..282
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     266..282
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             278..451
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(298..315)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(298..314)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(307..329)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             487..531
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(569..587)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      583..630
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(801..1592)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1593..1684)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1732..2470)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(2680..3762)
                     /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"
     primer_bind     3724..3743
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3763..3840)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3886..3902,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3886..3902,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcacacc atcaccacca tcacagcagc ggtctggaag ttctgtttca
      121 gggtaccgtc gacccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      181 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      241 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cggattcact
      301 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
      361 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
      421 ttcccaacag ttgcgcagcc tgaatggcga atggcgccaa gctttctaga ccagctcgag
      481 tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc acatggactc gtctactagc
      541 gcagcttaat taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg
      601 gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg
      661 gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt
      721 taacgaccct gccctgaacc gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg
      781 cttgaacgaa ttgttagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag
      841 tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga
      901 taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc
      961 cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg
     1021 gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc
     1081 gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc
     1141 tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc
     1201 agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat
     1261 tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca
     1321 acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc
     1381 aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc
     1441 agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt
     1501 acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga
     1561 gtcgatactt cggcgatcac cgcttccctc atactcttcc tttttcaata ttattgaagc
     1621 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     1681 caaatagcta gctcactcgg tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg
     1741 acacatacaa agttacccac agattccgtg gataagcagg ggactaacat gtgaggcaaa
     1801 acagcagggc cgcgccggtg gcgtttttcc ataggctccg ccctcctgcc agagttcaca
     1861 taaacagacg cttttccggt gcatctgtgg gagccgtgag gctcaaccat gaatctgaca
     1921 gtacgggcga aacccgacag gacttaaaga tccccaccgt ttccggcggg tcgctccctc
     1981 ttgcgctctc ctgttccgac cctgccgttt accggatacc tgttccgcct ttctccctta
     2041 cgggaagtgt ggcgctttct catagctcac acactggtat ctcggctcgg tgtaggtcgt
     2101 tcgctccaag ctgggctgta agcaagaact ccccgttcag cccgactgct gcgccttatc
     2161 cggtaactgt tcacttgagt ccaacccgga aaagcacggt aaaacgccac tggcagcagc
     2221 cattggtaac tgggagttcg cagaggattt gtttagctaa acacgcggtt gctcttgaag
     2281 tgtgcgccaa agtccggcta cactggaagg acagatttgg ttgctgtgct ctgcgaaagc
     2341 cagttaccac ggttaagcag ttccccaact gacttaacct tcgatcaaac cacctcccca
     2401 ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga aaaaaaggat ctcaagaaga
     2461 tcctttgatc ttttctactg aaccgctcta gatttcagtg caatttatct cttcaaatgt
     2521 agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc
     2581 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     2641 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2701 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2761 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     2821 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     2881 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     2941 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
     3001 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     3061 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     3121 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     3181 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     3241 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     3301 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     3361 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     3421 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     3481 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     3541 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     3601 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     3661 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     3721 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     3781 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     3841 cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta
     3901 ta
//
