LOCUS       sequence-54722-e        3971 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 3971)
  TITLE     OPPF-UK pOPIN vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3971)
  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            "Unpublished"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: pOPINRSE
FEATURES             Location/Qualifiers
     source          1..3971
                     /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             471..488
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             508..552
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(590..608)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      604..651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(881..1696)
                     /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(881..1696)
                     /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     1604..1623
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1697..1788)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1804..2553)
                     /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(1814..1831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2054..2073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2749..3831)
                     /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     3793..3812
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3832..3909)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(3955..3971,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(3955..3971,1..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 gctgtttaaa catcaccatc
      481 accatcacta agcttctcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag
      541 cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa
      601 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct
      661 caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag
      721 caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgacaag ctgacgaccg
      781 ggtctccgca agtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      841 ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca
      901 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
      961 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     1021 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     1081 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     1141 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     1201 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     1261 cgcggtcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     1321 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     1381 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     1441 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     1501 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     1561 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     1621 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     1681 gttgaatatg gctcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggcatgcagc
     1801 gctcttccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg
     1861 tgtcagctca ctcaaaagcg gtaatacggt tatccacaga atcaggggat aaagccggaa
     1921 agaacatgtg agcaaaaagc aaagcaccgg aagaagccaa cgccgcaggc gtttttccat
     1981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagccagag gtggcgaaac
     2041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2161 ctttctcata gctcacgctg ttggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccat tggtaactga
     2341 tttagaggac tttgtcttga agttatgcac ctgttaaggc taaactgaaa gaacagattt
     2401 tggtgagtgc ggtcctccaa cccacttacc ttggttcaaa gagttggtag ctcagcgaac
     2461 cttgagaaaa ccaccgttgg tagcggtggt ttttctttat ttatgagatg atgaatcaat
     2521 cggtctatca agtcaacgaa cagctattcc gttactctag atttcagtgc aatttatctc
     2581 ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc
     2641 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     2701 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     2761 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2821 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     2881 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     2941 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     3001 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     3061 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     3121 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     3181 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     3241 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     3301 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     3361 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     3421 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     3481 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     3541 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     3601 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     3661 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     3721 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     3781 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     3841 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     3901 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aaattaatac
     3961 gactcactat a
//
