LOCUS Exported 6783 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of conditionally active form of Cre recombinase (not as tightly regulated as pCAG-ERT2CreERT2). ACCESSION . VERSION . KEYWORDS pCAG-CreERT2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6783) 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 5. ():. PUBMED 17209010 REFERENCE 2 (bases 1 to 6783) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6783 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 386..661 /label=chicken beta-actin promoter intron 662..1670 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1594..1616 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1678..1697 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1731..1740 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1737..2765 /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="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE DGD" CDS 2778..3713 /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" primer_bind complement(3807..3826) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3872..3927 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3926..3945) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4286..4302) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4286..4302) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4299..4321) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4310..4326 /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(4334..4364) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4379..4400 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4459..4654 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4505..4640 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4567..4586 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4660..4794 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4697..4716) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4751..4770 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4862..4879) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5033..5621) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5113..5132) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5792..6652) /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 6415..6434 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6653..6757) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg 541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc 721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag 841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg 961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct 1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc 1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt 1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg 1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg 1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca 1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgagcc gccaccatgg 1741 ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca acgagtgatg 1801 aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct 1861 ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga 1921 aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt caggcgcgcg 1981 gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt catcgtcggt 2041 ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg cggatccgaa 2101 aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt 2161 tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata cgtaatctgg 2221 catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc aggatcaggg 2281 ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc agaacgaaaa 2341 cgctggttag caccgcaggt gtagagaagg cacttagcct gggggtaact aaactggtcg 2401 agcgatggat ttccgtctct ggtgtagctg atgatccgaa taactacctg ttttgccggg 2461 tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact cgcgccctgg 2521 aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac tctggtcaga 2581 gatacctggc ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg 2641 ctggagtttc aataccggag atcatgcaag ctggtggctg gaccaatgta aatattgtca 2701 tgaactatat ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg 2761 gcgatctcga gccatctgct ggagacatga gagctgccaa cctttggcca agcccgctca 2821 tgatcaaacg ctctaagaag aacagcctgg ccttgtccct gacggccgac cagatggtca 2881 gtgccttgtt ggatgctgag ccccccatac tctattccga gtatgatcct accagaccct 2941 tcagtgaagc ttcgatgatg ggcttactga ccaacctggc agacagggag ctggttcaca 3001 tgatcaactg ggcgaagagg gtgccaggct ttgtggattt gaccctccat gatcaggtcc 3061 accttctaga atgtgcctgg ctagagatcc tgatgattgg tctcgtctgg cgctccatgg 3121 agcacccagt gaagctactg tttgctccta acttgctctt ggacaggaac cagggaaaat 3181 gtgtagaggg catggtggag atcttcgaca tgctgctggc tacatcatct cggttccgca 3241 tgatgaatct gcagggagag gagtttgtgt gcctcaaatc tattattttg cttaattctg 3301 gagtgtacac atttctgtcc agcaccctga agtctctgga agagaaggac catatccacc 3361 gagtcctgga caagatcaca gacactttga tccacctgat ggccaaggca ggcctgaccc 3421 tgcagcagca gcaccagcgg ctggcccagc tcctcctcat cctctcccac atcaggcaca 3481 tgagtaacaa aggcatggag catctgtaca gcatgaagtg caagaacgtg gtgcccctct 3541 atgacctgct gctggaggcg gcggacgccc accgcctaca tgcgcccact agccgtggag 3601 gggcatccgt ggaggagacg gaccaaagcc acttggccac tgcgggctct acttcatcgc 3661 attccttgca aaagtattac atcacggggg aggcagaggg tttccctgcc acagcttgat 3721 agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat 3781 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc 3841 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata 3901 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta 3961 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgccatatg ctggctgcca 4021 tgaacaaagg tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc 4081 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt 4141 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct 4201 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatgaagat ccctcgacct 4261 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 4321 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 4381 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 4441 tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 4501 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 4561 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg 4621 aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat 4681 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 4741 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc 4801 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 4861 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 4921 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 4981 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 5041 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 5101 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 5161 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 5221 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 5281 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 5341 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 5401 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 5461 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 5521 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 5581 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 5641 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 5701 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 5761 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 5821 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 5881 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 5941 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 6001 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 6061 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 6121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 6181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 6241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 6301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 6361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 6421 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 6481 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 6541 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 6601 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 6661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 6721 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 6781 ctg //