LOCUS       sequence-233646-        6874 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 6874)
  AUTHORS   Ornitz DM, Moreadith RW, Leder P
  TITLE     Binary system for regulating transgene expression in mice: targeting
            int-2 gene expression with yeast GAL4/UAS control elements.
  JOURNAL   Proc Natl Acad Sci U S A 1991 Feb 1;88(3):698-702.
  PUBMED    1846961
REFERENCE   2  (bases 1 to 6874)
  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: "Proc Natl 
            Acad Sci U S A"; date: "1991-02-1"; volume: "88(3)"; pages: 
            "698-702"
COMMENT     Sequence Label: MMTV-SV40-Bssk
FEATURES             Location/Qualifiers
     source          1..6874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     271..290
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     intron          3416..3481
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3611..3631
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3903..4037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3940..3959)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3994..4013
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4034..4050)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4034..4050)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4087..4105)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4126..4142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4126..4142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4139..4161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4150..4166
                     /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)."
     promoter        complement(join(4174..4180,4181..4198,4199..4204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4219..4240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4357..4374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4528..5116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4608..4627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5287..6078,6079..6147))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5910..5929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6148..6252)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6278..6733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6415..6436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6627..6646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6860..6874,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6874,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg ggtaccgggc
       61 cccccctcga ggtcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
      121 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
      181 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
      241 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
      301 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcccaatgat
      361 agagatttta ctgctctagt tccccataca gaattgtttc gcttagttgc agcctcaaga
      421 tatcttattc tcaaaaggcc aggatttcaa gaacatgaca tgattcctac atctgcctgt
      481 gttacttacc cttatgccat attattagga ttacctcagc taatagatat agagaaaaga
      541 ggatctactt ttcatatttc ctgttcttct tgtagattga ctaattgttt agattcttct
      601 gcctacgact atgcagcgat catagtcaag aggccgccat acgtgctgct acctgtagat
      661 attggtgatg aaccatggtt tgatgattct gccattcaaa cctttaggta tgccacagat
      721 ttaattcgag ccaagcgatt cgtcgctgcc attattctgg gcatatctgc tttaattgct
      781 attatcactt cctttgctgt agctactact gctttagtta aggagatgca aactgctacg
      841 tttgttaata atcttcatag gaatgttaca ttagccttat ctgaacaaag aataatagat
      901 ttaaaattag aagctagact taatgcttta gaagaagtag ttttagagtt gggacaagat
      961 gtggcaaact taaagaccag aatgtccacc aggtgtcatg caaattatga ttttatctgc
     1021 gttacacctt taccatataa tgcttctgag agctgggaaa gaaccaaagc tcatttattg
     1081 ggcatttgga atgacaatga gatttcatat aacatacaag aattaaccaa cctgattagt
     1141 gatatgagca aacaacatat tgacacagtg gacctcagtg gcttggctca gtcctttgcc
     1201 aatggagtaa aggctttaaa tccattagat tggacacaat atttcatttt tataggtgtt
     1261 ggagccctgc ttttagtcat agtgcttatg attttcccca ttgttttcca gtgccttgcg
     1321 aagagccttg accaagtgca gtcagatctt aacgtgcttc ttttaaaaaa gaaaaaaggg
     1381 ggaaatgccg cgcctgcagc agaaatggtt gaactcccga gagtgtccta cacctagggg
     1441 agaagcagcc aaggggttgt ttcccaccaa ggacgacccg tctgcgcaca aacggatgag
     1501 cccatcagac aaagacatat tcattctctg ctgcaaactt ggcatagctc tgctttgcct
     1561 ggggctattg ggggaagttg cggttcgtgc tcgcagggct ctcacccttg actcttttaa
     1621 tagctcttct gtgcaagatt acaatctaaa caattcggag aactcgacct tcctcctgag
     1681 gcaaggacca cagccaactt cctcttacaa gccgcatcga ttttgtcctt cagaaataga
     1741 aataagaatg cttgctaaaa attatatttt taccaataag accaatccaa taggtagatt
     1801 attagttact atgttaagaa atgaatcatt atcttttagt actattttta ctcaaattca
     1861 gaagttagaa atgggaatag aaaatagaaa gagacgctca acctcaattg aagaacaggt
     1921 gcaaggacta ttgaccacag gcctagaagt aaaaaaggga aaaaagagtg tttttgtcaa
     1981 aataggagac aggtggtggc aaccagggac ttatagggga ccttacatct acagaccaac
     2041 agatgccccc ttaccatata caggaagata tgacttaaat tgggataggt gggttacagt
     2101 caatggctat aaagtgttat atagatccct cccttttcgt gaaagactcg ccagagctag
     2161 acctccttgg tgtatgttgt ctcaagaaga aaaagacgac atgaaacaac aggtacatga
     2221 ttatatttat ctaggaacag gaatgcactt ttggggaaag attttccata ccaaggaggg
     2281 gacagtggct ggactaatag aacattattc tgcaaaaact tatggcatga gttattatga
     2341 atagccttta ttggcccaac cttgcggttc ccagggctta agtaagtttt tggttacaaa
     2401 ctgttcttaa aacgaggatg tgagacaagt ggtttcctga cttggtttgg tatcaaaggt
     2461 tctgatctga gctctgagtg ttctattttc ctatgttctt ttggaattta tccaaatctt
     2521 atgtaaatgc ttatgtaaac caagatataa aagagtgctg attttttgag taaacttgca
     2581 acagtcctaa cattcacctc ttgtgtgttt gtgtctgttc gccatcccgt ctccgctcgt
     2641 cacttatcct tcactttcca gagggtcccc ccgcagaccc cggcgtagag gatccgcacc
     2701 cttgatgact ccgtctgaat ttttggtttc agtttggtac cgaagctgcg cggcgcgtct
     2761 gcttgttact tgtttgactg ttggaattgt ttgtcttctt tgtgacctga ctgtggtttt
     2821 ctggacgtgt tgtgtctgtt agtgtctttt tgacttttgt ttcgtgtttg aatttggact
     2881 gacgactgtg tttaaaatct tagaccgacg actgtgtttg aaatcatgaa actgtttgct
     2941 ttgttcgtcg aagagtttta cttggtcccc ttaacgctta gtgagtaaga aacttaattt
     3001 tgtagacccc gctctagtgg cagtgtgttg gttgatagcc aaagttaatt tttaaaacat
     3061 agtgttttgg gggttgggga tttagctcag tgatagagct cttgcctagc aagcgcaagg
     3121 ccctgggttc ggtccccagc tctgaaaaaa aggaaagaga aacaaaacaa aaacatatag
     3181 tgttttatct gtgcttatgc ccgcagcccg agccgcaccc gccgcggacg gagcccatgc
     3241 gcgggcccag tcggcgcccg tccgcgcccc cgccctgccc cggccccggc ccccaagctt
     3301 gatatcgaat tcctgcaggt cgcggccgcg actctagagg atctttgtga aggaacctta
     3361 cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa
     3421 tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta
     3481 gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa
     3541 cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca
     3601 ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt
     3661 gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta
     3721 caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat atttgatgta
     3781 tagtgccttg actagagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3841 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3901 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3961 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4021 cttatcatgt ctggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt
     4081 ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc
     4141 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4201 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4261 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4321 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4381 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4441 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5221 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5281 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5401 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5581 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6121 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6241 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     6301 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6361 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     6421 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6481 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     6541 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     6601 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     6661 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     6721 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6781 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6841 tgggtaacgc cagggttttc ccagtcacga cgtt
//
