LOCUS       sequence-7770-ex        6803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of enhanced form of Flp recombinase fused to 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 16;104(3):1027-32. doi: 
            10.1073/pnas.0610155104
  PUBMED    17209010
REFERENCE   2  (bases 1 to 6803)
  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-Flpe:GFP
FEATURES             Location/Qualifiers
     source          1..6803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3021..3023,3024..3026,3027..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             join(3021..3023,3024..3026,3027..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(join(4354..4360,4361..4378,4379..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"
     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(join(5812..6603,6604..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
//
