LOCUS Exported 6803 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of enhanced form of Flp recombinase fused to GFP. ACCESSION . VERSION . KEYWORDS pCAG-Flpe:GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6803) 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 6803) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6803 /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" regulatory 3015..3024 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3021..3740 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3021..3737 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3066..3087) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3327..3346) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3674..3695 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3827..3846) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3892..3947 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3946..3965) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4306..4322) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4306..4322) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4319..4341) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4330..4346 /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(4354..4384) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4399..4420 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4479..4674 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4525..4660 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4587..4606 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4680..4814 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4717..4736) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4771..4790 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4882..4899) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5053..5641) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5133..5152) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5812..6672) /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 6435..6454 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6673..6777) /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 gcataggacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc 3061 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 3121 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 3181 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc 3241 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 3301 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 3361 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 3421 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca 3481 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg 3541 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 3601 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 3661 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 3721 tggacgagct gtacaagtaa agcggccgca ctcctcaggt gcaggctgcc tatcagaagg 3781 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg 3841 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga 3901 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat 3961 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat 4021 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa 4081 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt 4141 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt 4201 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc 4261 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc 4321 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 4381 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 4441 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc 4501 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 4561 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 4621 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta 4681 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4741 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4801 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 4861 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 5041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5221 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta 5281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5461 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 5521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 5641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 5701 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 5761 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 5821 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 5881 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 5941 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 6001 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 6061 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 6121 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 6181 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 6241 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 6301 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 6361 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 6421 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 6481 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 6541 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 6601 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 6661 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 6721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 6781 atttccccga aaagtgccac ctg //