LOCUS       sequence-373729-        7608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  green biosensor for cAMP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7608)
  AUTHORS   Wang L, Wu C, Peng W, Zhou Z, Zeng J, Li X, Yang Y, Yu S, Zou Y, 
            Huang M, Liu C, Chen Y, Li Y, Ti P, Liu W, Gao Y, Zheng W, Zhong H, 
            Gao S, Lu Z, Ren PG, Ng HL, He J, Chen S, Xu M, Li Y, Chu J
  TITLE     A high-performance genetically encoded fluorescent indicator for in 
            vivo cAMP imaging.
  JOURNAL   Nat Commun. 2022 Sep 12;13(1):5363. doi: 10.1038/s41467-022-32994-7.
  PUBMED    36097007
REFERENCE   2  (bases 1 to 7608)
  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"; journalName: "Nat 
            Commun."; date: "2022-09-12"; volume: "13(1):5363. doi"; pages: " 
            10.1038/s41467-022-32994-7"
COMMENT     Sequence Label: pCAG-G-Flamp1
FEATURES             Location/Qualifiers
     source          1..7608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        92..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          671..1687
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1611..1633
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1695..1714
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1789..1798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1804..1821
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1825..1857
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1861..1869,1870..1884)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     2325..2346
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2452..2473)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3112..3141
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3157..3174
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3197..3214)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3203..3427
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3473..3901
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3560..3579)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3770..3791
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3910..3930)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3915..4244
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4095..4230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4157..4176
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4311..5105
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4365..4384)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4975..4994
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5281..5402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5318..5337)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5372..5391
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5451..5467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5451..5467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5464..5486)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5475..5491
                     /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(5499..5505,5506..5523,5524..5529))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5544..5565
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5682..5699)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5853..6441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5933..5952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6612..7403,7404..7472))
                     /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     7235..7254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7473..7577)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcaatt ctgcaggaat ctagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
      421 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
      481 tattttgtgc agcgatgggg gcgggggggg ggggggsgcg ccrggsgggg gggggggggg
      541 gggggggggg gggggggsgr ggsggrgagg kgcggcggca gccaatcaga gcggcgcgct
      601 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
      661 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
      721 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
      781 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
      841 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
      901 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg
      961 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag
     1021 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt
     1081 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc
     1141 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg
     1201 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc
     1261 gggcggggcg gggccgcctc gggccggggr gggctcgggg grggggcgcg gcggcccccg
     1321 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg
     1381 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg
     1441 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat
     1501 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg
     1561 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
     1621 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     1681 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     1741 gatttgatac cgcgggcccg ggatcagctt ggtaccgagc tcggatccgc caccatgggt
     1801 tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg
     1861 gatctgtacg acgatgacga taaggatccg atgggcttct atcaagaagt ccgtcgcggg
     1921 gatttcgtcc gcaattggca attggtcgcc gccgtgccgt tgtttcagaa gctcggcccg
     1981 gccgtgctgg tcgagatcgt gcgcgcctta agagcccgca cggtgccggc gggcgccgtg
     2041 atctgccgca ttggcgagcc cggcgatcgg atgttcttcg tcgtggaggg gagcgtcagc
     2101 gtcgcgacga attgggggaa cgtctatatc acagccgaca agcagaagaa cggcatcaag
     2161 gcgaacttca agatccgcca caacgttgag ggcggcggcg tgcagctcgc ctaccactac
     2221 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2281 gtgcagtcca aactttcgaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2341 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga
     2401 gggagcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2461 ctggacggcg acgtaaacgg ccacaagttc agcgtgcgtg gcgagggcga gggcgatgcc
     2521 accaatggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2581 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcgcacgcta ccccgaccac
     2641 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacatcca ggagcgcacc
     2701 atcgttttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac
     2761 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2821 gggcacaagc tggagtacaa ccgcgtgaat ccggtggagc ttggccctgg cgccttcttc
     2881 ggcgagatgg cgctgatcag cggcgaaccg cgtgttgcga ccgtcagcgc cgcaacgacg
     2941 gtctcactcc tgtcgctgca ttcggcggat ttccagatgt tgtgcagcag cagcccggag
     3001 atcgcggaaa tcttccgcaa gaccgcgctc gagcgtcgcg gcgctgcggc gagcgcttaa
     3061 tgaattctgc agatatccag cacagtggcg gccgctcgag gtcacccatt cgaacaaaaa
     3121 ctcatctcag aagaggatct gaatatgcat accggtcatc atcaccatca ccattgagtt
     3181 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3241 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3301 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3361 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3421 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3481 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3541 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3601 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3661 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3721 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3781 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3841 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3901 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3961 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4021 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4081 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4141 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4201 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4261 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4321 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4381 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4441 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4501 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4561 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4621 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4681 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4741 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4801 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4861 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4921 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4981 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5041 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5101 tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca
     5161 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5221 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5281 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5341 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5401 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5461 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5521 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5581 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5641 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5701 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5761 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5821 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5881 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5941 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6001 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6061 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6121 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6181 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6241 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6301 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6361 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6421 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6481 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6541 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6601 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6661 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6721 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6781 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6841 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6901 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6961 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7021 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7081 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7141 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7201 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7261 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7321 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7381 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7441 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7501 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7561 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
