LOCUS       sequence-326722-        7740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of conditionally active form of Cre recombinase
            (activated in response to tamoxifen).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7740)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Controlled expression of transgenes introduced by in vivo 
            electroporation.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):1027-32. doi: 
            10.1073/pnas.0610155104
  PUBMED    17209010
REFERENCE   2  (bases 1 to 7740)
  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"; doi: "10.1073/pnas"; 
            journalName: "Proc Natl Acad Sci U S A"; date: "2007-01-16- 16"; 
            volume: "104"; issue: "3"; pages: "1027-32"
COMMENT     Sequence Label: pCAG-ERT2CreERT2
FEATURES             Location/Qualifiers
     source          1..7740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1744..1753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1753..2688
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2698..3720
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="NLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLS
                     VCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPR
                     PSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLG
                     IAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERW
                     ISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRY
                     LAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLEDG
                     D"
     CDS             3733..4668
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             3781..4674
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     primer_bind     complement(4762..4781)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4827..4882
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4881..4900)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5243..5259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5243..5259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5256..5278)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5267..5283
                     /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(5291..5297,5298..5315,5316..5321))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5336..5357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5416..5611
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5462..5597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5617..5751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5654..5673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5708..5727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5819..5836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5990..6578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6070..6089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6749..7540,7541..7609))
                     /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     7372..7391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7610..7714)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccrggsggg gsggggcggg gcgrggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcccgggtga
     1741 gccgccacca tggctggaga catgagagct gccaaccttt ggccaagccc gctcatgatc
     1801 aaacgctcta agaagaacag cctggccttg tccctgacgg ccgaccagat ggtcagtgcc
     1861 ttgttggatg ctgagccccc catactctat tccgagtatg atcctaccag acccttcagt
     1921 gaagcttcga tgatgggctt actgaccaac ctggcagaca gggagctggt tcacatgatc
     1981 aactgggcga agagggtgcc aggctttgtg gatttgaccc tccatgatca ggtccacctt
     2041 ctagaatgtg cctggctaga gatcctgatg attggtctcg tctggcgctc catggagcac
     2101 ccagtgaagc tactgtttgc tcctaacttg ctcttggaca ggaaccaggg aaaatgtgta
     2161 gagggcatgg tggagatctt cgacatgctg ctggctacat catctcggtt ccgcatgatg
     2221 aatctgcagg gagaggagtt tgtgtgcctc aaatctatta ttttgcttaa ttctggagtg
     2281 tacacatttc tgtccagcac cctgaagtct ctggaagaga aggaccatat ccaccgagtc
     2341 ctggacaaga tcacagacac tttgatccac ctgatggcca aggcaggcct gaccctgcag
     2401 cagcagcacc agcggctggc ccagctcctc ctcatcctct cccacatcag gcacatgagt
     2461 aacaaaggca tggagcatct gtacagcatg aagtgcaaga acgtggtgcc cctctatgac
     2521 ctgctgctgg aggcggcgga cgcccaccgc ctacatgcgc ccactagccg tggaggggca
     2581 tccgtggagg agacggacca aagccacttg gccactgcgg gctctacttc atcgcattcc
     2641 ttgcaaaagt attacatcac gggggaggca gagggtttcc ctgccacagc tgtcgacaat
     2701 ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag tgatgaggtt
     2761 cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca tacctggaaa
     2821 atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa ccggaaatgg
     2881 tttcccgcag aacctgaaga tgttcgcgat tatcttctat atcttcaggc gcgcggtctg
     2941 gcagtaaaaa ctatccagca acatttgggc cagctaaaca tgcttcatcg tcggtccggg
     3001 ctgccacgac caagtgacag caatgctgtt tcactggtta tgcggcggat ccgaaaagaa
     3061 aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac tgatttcgac
     3121 caggttcgtt cactcatgga aaatagcgat cgctgccagg atatacgtaa tctggcattt
     3181 ctggggattg cttataacac cctgttacgt atagccgaaa ttgccaggat cagggttaaa
     3241 gatatctcac gtactgacgg tgggagaatg ttaatccata ttggcagaac gaaaacgctg
     3301 gttagcaccg caggtgtaga gaaggcactt agcctggggg taactaaact ggtcgagcga
     3361 tggatttccg tctctggtgt agctgatgat ccgaataact acctgttttg ccgggtcaga
     3421 aaaaatggtg ttgccgcgcc atctgccacc agccagctat caactcgcgc cctggaaggg
     3481 atttttgaag caactcatcg attgatttac ggcgctaagg atgactctgg tcagagatac
     3541 ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc gagatatggc ccgcgctgga
     3601 gtttcaatac cggagatcat gcaagctggt ggctggacca atgtaaatat tgtcatgaac
     3661 tatatccgta acctggatag tgaaacaggg gcaatggtgc gcctgctgga agatggcgat
     3721 ctcgagccat ctgctggaga catgagagct gccaaccttt ggccaagccc gctcatgatc
     3781 aaacgctcta agaagaacag cctggccttg tccctgacgg ccgaccagat ggtcagtgcc
     3841 ttgttggatg ctgagccccc catactctat tccgagtatg atcctaccag acccttcagt
     3901 gaagcttcga tgatgggctt actgaccaac ctggcagaca gggagctggt tcacatgatc
     3961 aactgggcga agagggtgcc aggctttgtg gatttgaccc tccatgatca ggtccacctt
     4021 ctagaatgtg cctggctaga gatcctgatg attggtctcg tctggcgctc catggagcac
     4081 ccagtgaagc tactgtttgc tcctaacttg ctcttggaca ggaaccaggg aaaatgtgta
     4141 gagggcatgg tggagatctt cgacatgctg ctggctacat catctcggtt ccgcatgatg
     4201 aatctgcagg gagaggagtt tgtgtgcctc aaatctatta ttttgcttaa ttctggagtg
     4261 tacacatttc tgtccagcac cctgaagtct ctggaagaga aggaccatat ccaccgagtc
     4321 ctggacaaga tcacagacac tttgatccac ctgatggcca aggcaggcct gaccctgcag
     4381 cagcagcacc agcggctggc ccagctcctc ctcatcctct cccacatcag gcacatgagt
     4441 aacaaaggca tggagcatct gtacagcatg aagtgcaaga acgtggtgcc cctctatgac
     4501 ctgctgctgg aggcggcgga cgcccaccgc ctacatgcgc ccactagccg tggaggggca
     4561 tccgtggagg agacggacca aagccacttg gccactgcgg gctctacttc atcgcattcc
     4621 ttgcaaaagt attacatcac gggggaggca gagggtttcc ctgccacagc ttgatagcgg
     4681 ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct
     4741 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     4801 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     4861 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     4921 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     4981 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     5041 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     5101 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     5161 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     5221 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5281 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5341 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5401 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5461 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5521 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5581 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
     5641 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5701 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
     5761 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5821 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5881 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5941 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6001 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6061 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6121 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6181 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6241 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6301 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6361 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6421 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6481 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6541 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6601 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6661 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6721 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6781 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6841 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6901 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6961 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7021 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7081 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7141 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7201 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7261 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7321 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7381 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7441 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7501 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7561 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7621 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7681 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
