LOCUS       sequence-54712-e        3950 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 3950)
  TITLE     OPPF-UK pOPIN vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3950)
  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            "Unpublished"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: pOPINRSF
FEATURES             Location/Qualifiers
     source          1..3950
                     /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(860..1675)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(860..1675)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1583..1602
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1676..1767)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1783..2532)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(1793..1810)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2033..2052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2728..3810)
                     /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     3772..3791
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3811..3888)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3934..3950,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3934..3950,1..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 gacgacaagc tgacgaccgg gtctccgcaa gtggcacttt
      781 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      841 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
      901 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      961 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     1021 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     1081 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     1141 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     1201 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcggtcgctg ttaaaaggac
     1261 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1321 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     1381 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1441 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1501 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1561 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1621 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct
     1681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1741 ttgaatgtat ttagaaaaat aaacaaatag gcatgcagcg ctcttccgct tcctcgctca
     1801 ctgactcgct acgctcggtc gttcgactgc ggcgagcggt gtcagctcac tcaaaagcgg
     1861 taatacggtt atccacagaa tcaggggata aagccggaaa gaacatgtga gcaaaaagca
     1921 aagcaccgga agaagccaac gccgcaggcg tttttccata ggctccgccc ccctgacgag
     1981 catcacaaaa atcgacgctc aagccagagg tggcgaaacc cgacaggact ataaagatac
     2041 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2101 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2161 tggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2221 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2281 cacgacttat cgccactggc agcagccatt ggtaactgat ttagaggact ttgtcttgaa
     2341 gttatgcacc tgttaaggct aaactgaaag aacagatttt ggtgagtgcg gtcctccaac
     2401 ccacttacct tggttcaaag agttggtagc tcagcgaacc ttgagaaaac caccgttggt
     2461 agcggtggtt tttctttatt tatgagatga tgaatcaatc ggtctatcaa gtcaacgaac
     2521 agctattccg ttactctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
     2581 cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag
     2641 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     2701 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2761 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2821 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     2881 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     2941 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     3001 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     3061 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     3121 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     3181 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     3241 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     3301 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     3361 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     3421 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     3481 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     3541 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     3601 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     3661 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     3721 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     3781 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     3841 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     3901 gctctccctt atgcgactcc tgcattagga aattaatacg actcactata
//
