LOCUS Exported 7029 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCAG-FlpeERT2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7029) 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 7029) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7029 /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" CDS 1737..3005 /codon_start=1 /product="site-specific recombinase" /label=FLP /note="FLP is a site-specific recombinase from Saccharomyces cerevisiae. Recombination occurs at FRT sequences." /translation="MSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAAELTYLCWMI THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI IPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR SYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR YPAWNGIISQEVLDYLSSYINRRI" CDS 3024..3959 /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(4053..4072) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4118..4173 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4172..4191) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4532..4548) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4532..4548) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4545..4567) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4556..4572 /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(4580..4610) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4625..4646 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4705..4900 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4751..4886 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4813..4832 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4906..5040 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4943..4962) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4997..5016 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5108..5125) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5279..5867) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5359..5378) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6038..6898) /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 6661..6680 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6899..7003) /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 gccaccatga 1741 gccaatttga tatattatgt aaaacaccac ctaaggtcct ggttcgtcag tttgtggaaa 1801 ggtttgaaag accttcaggg gaaaaaatag catcatgtgc tgctgaacta acctatttat 1861 gttggatgat tactcataac ggaacagcaa tcaagagagc cacattcatg agctataata 1921 ctatcataag caattcgctg agtttcgata ttgtcaacaa atcactccag tttaaataca 1981 agacgcaaaa agcaacaatt ctggaagcct cattaaagaa attaattcct gcttgggaat 2041 ttacaattat tccttacaat ggacaaaaac atcaatctga tatcactgat attgtaagta 2101 gtttgcaatt acagttcgaa tcatcggaag aagcagataa gggaaatagc cacagtaaaa 2161 aaatgcttaa agcacttcta agtgagggtg aaagcatctg ggagatcact gagaaaatac 2221 taaattcgtt tgagtatacc tcgagattta caaaaacaaa aactttatac caattcctct 2281 tcctagctac tttcatcaat tgtggaagat tcagcgatat taagaacgtt gatccgaaat 2341 catttaaatt agtccaaaat aagtatctgg gagtaataat ccagtgttta gtgacagaga 2401 caaagacaag cgttagtagg cacatatact tctttagcgc aaggggtagg atcgatccac 2461 ttgtatattt ggatgaattt ttgaggaact ctgaaccagt cctaaaacga gtaaatagga 2521 ccggcaattc ttcaagcaac aaacaggaat accaattatt aaaagataac ttagtcagat 2581 cgtacaacaa ggctttgaag aaaaatgcgc cttatccaat ctttgctata aagaatggcc 2641 caaaatctca cattggaaga catttgatga cctcatttct gtcaatgaag ggcctaacgg 2701 agttgactaa tgttgtggga aattggagcg ataagcgtgc ttctgccgtg gccaggacaa 2761 cgtatactca tcagataaca gcaatacctg atcactactt cgcactagtt tctcggtact 2821 atgcatatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg 2881 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg 2941 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac 3001 gcatatgcgt acgcggatcc tctgctggag acatgagagc tgccaacctt tggccaagcc 3061 cgctcatgat caaacgctct aagaagaaca gcctggcctt gtccctgacg gccgaccaga 3121 tggtcagtgc cttgttggat gctgagcccc ccatactcta ttccgagtat gatcctacca 3181 gacccttcag tgaagcttcg atgatgggct tactgaccaa cctggcagac agggagctgg 3241 ttcacatgat caactgggcg aagagggtgc caggctttgt ggatttgacc ctccatgatc 3301 aggtccacct tctagaatgt gcctggctag agatcctgat gattggtctc gtctggcgct 3361 ccatggagca cccagtgaag ctactgtttg ctcctaactt gctcttggac aggaaccagg 3421 gaaaatgtgt agagggcatg gtggagatct tcgacatgct gctggctaca tcatctcggt 3481 tccgcatgat gaatctgcag ggagaggagt ttgtgtgcct caaatctatt attttgctta 3541 attctggagt gtacacattt ctgtccagca ccctgaagtc tctggaagag aaggaccata 3601 tccaccgagt cctggacaag atcacagaca ctttgatcca cctgatggcc aaggcaggcc 3661 tgaccctgca gcagcagcac cagcggctgg cccagctcct cctcatcctc tcccacatca 3721 ggcacatgag taacaaaggc atggagcatc tgtacagcat gaagtgcaag aacgtggtgc 3781 ccctctatga cctgctgctg gaggcggcgg acgcccaccg cctacatgcg cccactagcc 3841 gtggaggggc atccgtggag gagacggacc aaagccactt ggccactgcg ggctctactt 3901 catcgcattc cttgcaaaag tattacatca cgggggaggc agagggtttc cctgccacag 3961 cttgatagcg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg 4021 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg 4081 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt 4141 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat 4201 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc catatgctgg 4261 ctgccatgaa caaaggtggc tataaagagg tcatcagtat atgaaacagc cccctgctgt 4321 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt 4381 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt 4441 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat gaagatccct 4501 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 4561 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 4621 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 4681 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct 4741 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 4801 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 4861 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca 4921 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 4981 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc 5041 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 5101 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 5161 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 5221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 5281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 5341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 5401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 5461 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 5521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 5581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 5641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 5701 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 5761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 5821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 5881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 5941 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 6001 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 6061 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 6121 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 6181 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 6241 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 6301 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 6361 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 6421 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 6481 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 6541 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 6601 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 6661 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 6721 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 6781 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 6841 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 6901 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 6961 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 7021 tgccacctg //