LOCUS       sequence-54727-e        5012 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 5012)
  TITLE     OPPF-UK pOPIN vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5012)
  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            "Unpublished"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: pOPINCDM
FEATURES             Location/Qualifiers
     source          1..5012
                     /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"
     CDS             104..1204
                     /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
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1178..1201
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    1278..1299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1314..1319,1320..1337,1338..1344)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1352..1368
                     /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     1357..1379
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1376..1392
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1376..1392
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1388..1561
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(1408..1425)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1408..1424)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1417..1439)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1597..1641
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1679..1697)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1693..1740
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1911..2702)
                     /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(2703..2794)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2842..3580)
                     /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(3790..4872)
                     /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     4834..4853
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4873..4950)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(4996..5012,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4996..5012,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcacacc atcaccacca tcacagcagc ggcatgaaaa tcgaagaagg
      121 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      181 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      241 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      301 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      361 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      421 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      481 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      541 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      601 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      661 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      721 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
      781 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
      841 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
      901 tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
      961 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1021 gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccactatgg aaaacgccca
     1081 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1141 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1201 gactagcagc ggtctggaag ttctgtttca gggtaccgtc gacccgactg gaaagcgggc
     1261 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     1321 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     1381 aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga
     1441 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1501 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1561 atggcgccaa gctttctaga ccagctcgag tctggtaaag aaaccgctgc tgcgaaattt
     1621 gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc
     1681 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1741 ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     1801 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     1861 tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca ttatttgccg
     1921 actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc
     1981 gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc
     2041 tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt
     2101 ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg
     2161 ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga
     2221 tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta
     2281 tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag
     2341 atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga
     2401 taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc
     2461 tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt
     2521 gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg
     2581 ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc
     2641 tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc
     2701 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2761 tacatatttg aatgtattta gaaaaataaa caaatagcta gctcactcgg tcgctacgct
     2821 ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa agttacccac agattccgtg
     2881 gataagcagg ggactaacat gtgaggcaaa acagcagggc cgcgccggtg gcgtttttcc
     2941 ataggctccg ccctcctgcc agagttcaca taaacagacg cttttccggt gcatctgtgg
     3001 gagccgtgag gctcaaccat gaatctgaca gtacgggcga aacccgacag gacttaaaga
     3061 tccccaccgt ttccggcggg tcgctccctc ttgcgctctc ctgttccgac cctgccgttt
     3121 accggatacc tgttccgcct ttctccctta cgggaagtgt ggcgctttct catagctcac
     3181 acactggtat ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta agcaagaact
     3241 ccccgttcag cccgactgct gcgccttatc cggtaactgt tcacttgagt ccaacccgga
     3301 aaagcacggt aaaacgccac tggcagcagc cattggtaac tgggagttcg cagaggattt
     3361 gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa agtccggcta cactggaagg
     3421 acagatttgg ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag ttccccaact
     3481 gacttaacct tcgatcaaac cacctcccca ggtggttttt tcgtttacag ggcaaaagat
     3541 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctcta
     3601 gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa
     3661 gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     3721 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     3781 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3841 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     3901 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     3961 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     4021 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc
     4081 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     4141 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     4201 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     4261 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     4321 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     4381 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     4441 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     4501 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     4561 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     4621 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     4681 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     4741 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     4801 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     4861 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     4921 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     4981 cctgcattag gaaattaata cgactcacta ta
//
