LOCUS       sequence-266281-        7598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cGMP FRET biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7598)
  AUTHORS   Ros O, Zagar Y, Ribes S, Baudet S, Loulier K, Couvet S, Ladarre D, 
            Aghaie A, Louail A, Petit C, Mechulam Y, Lenkei Z, Nicol X
  TITLE     SponGee: A Genetic Tool for Subcellular and Cell-Specific cGMP 
            Manipulation.
  JOURNAL   Cell Rep. 2019 Jun 25;27(13):4003-4012.e6. doi: 
            10.1016/j.celrep.2019.05.102.
  PUBMED    31242429
REFERENCE   2  (bases 1 to 7598)
  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: 
            "10.1016/j.celrep.2019.05"; journalName: "Cell Rep"; date: 
            "2019-06-25- 25"; volume: "27"; issue: "13"; pages: "4003-4012.e6"
COMMENT     Sequence Label: pCX-ThPDE5VV
FEATURES             Location/Qualifiers
     source          1..7598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..660
                     /label=chicken beta-actin promoter
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1818..1820,1821..1823,1824..2336)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK
                     ANFKIRHNIE"
     CDS             2985..3719
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     CDS             join(3777..3779,3780..3782,3783..4493)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4619..4638)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4684..4739
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4738..4757)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5100..5116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5100..5116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5113..5135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5124..5140
                     /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(5148..5154,5155..5172,5173..5178))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5193..5214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5273..5468
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5319..5454
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5474..5608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5511..5530)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5565..5584
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5676..5693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5847..6435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5927..5946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6606..7397,7398..7466))
                     /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     7229..7248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7467..7571)
                     /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 tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
      541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ctgataaact
     1741 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca
     1801 gcacagtggc ggccgcaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1861 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1921 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1981 ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct
     2041 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2101 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2161 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2221 gcaacatcct ggggcacaag ctggagtaca actacatcag cgacaacgtc tatatcaccg
     2281 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     2341 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2401 tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga
     2461 agcgcgatca catggtcctg ctggagttcg tgaccgccgc ccgatgggaa ttagtgaagg
     2521 atatttctag tcatttggat gtcacagcct tatgtcacaa aattttcttg catatccatg
     2581 gactgatatc tgctgaccgc tattccctgt tccttgtctg tgaagacagc tccaatgaca
     2641 agtttcttat cagccgcctc tttgatgttg ctgaaggttc aacactggaa gaagtttcaa
     2701 ataactgtat ccgcttagaa tggaacaaag gcattgtggg acatgtggca gcgcttggtg
     2761 agcccttgaa catcaaagat gcatatgagg atcctcggtt caatgcagaa gttgaccaaa
     2821 ttacaggcta caagacacaa agcattcttt gtatgccaat taagaatcat agggaagagg
     2881 ttgttggtgt agcccaggcc atcaacaaga aatcaggaaa cggtgggaca tttactgaaa
     2941 aagatgaaaa ggactttgct gcttatttgg cagctagcga gctcatggac ggcggcgtgc
     3001 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3061 acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3121 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3181 acaagggtgg cagcggtggc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     3241 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3301 gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc
     3361 tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc
     3421 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3481 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3541 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3601 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3661 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagc
     3721 cgaattttgt cttcctgatc ggcgccgcag gaatactctt cgtatctagc gagctcatgg
     3781 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     3841 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     3901 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     3961 tgaccactct gggctacggc ctacagtgct tcgcccgcta ccccgaccac atgaagcagc
     4021 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     4081 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     4141 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     4201 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
     4261 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     4321 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     4381 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     4441 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagccgaatt
     4501 ccctcgaggt taacgcggcg cgcctctaat ctagcgaatt cactcctcag gtgcaggctg
     4561 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     4621 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     4681 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     4741 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     4801 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     4861 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     4921 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     4981 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     5041 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     5101 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5161 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5221 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     5281 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5341 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5401 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5461 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5521 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5581 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     5641 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5701 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5761 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5821 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5881 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5941 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6001 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6061 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6121 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6181 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6241 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6301 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6361 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6421 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6481 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6541 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6601 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6661 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6721 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6781 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6841 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6901 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6961 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7021 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7081 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7141 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7201 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7261 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7321 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7381 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7441 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7501 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7561 ggggttccgc gcacatttcc ccgaaaagtg ccacctgg
//
