LOCUS       sequence-72719-e        5664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for cGMP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  AUTHORS   Bhargava Y, Hampden-Smith K, Chachlaki K, Wood KC, Vernon J, 
            Allerston CK, Batchelor AM, Garthwaite J
  TITLE     Improved genetically-encoded, FlincG-type fluorescent biosensors for
            neural cGMP imaging.
  JOURNAL   Front Mol Neurosci. 2013 Sep 24;6:26. doi: 10.3389/fnmol.2013.00026.
  PUBMED    24068983
REFERENCE   2  (bases 1 to 5664)
  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: "Front Mol 
            Neurosci."; date: "2013-09-24"; volume: "6:26. doi"; pages: " 
            10.3389/fnmol.2013.00026"
COMMENT     Sequence Label: pC1-FGB (FlincG2)
FEATURES             Location/Qualifiers
     source          1..5664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1664..1685
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1791..1812)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2052..2071)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2452..2573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2489..2508)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2543..2562
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2580..3035)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2717..2738)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2929..2948
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3062..3166
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3168..3525
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3168..3359
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3191..3211)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3376..3511
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3560..4354
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3560..4354
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3614..3633)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4224..4243
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4542..4561)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4586..4633
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4962..5550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5451..5470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgact
      601 ggatccggac catggcactc cgacaggcat tccggaagtt caccaaatcc gaaaggtcca
      661 aggatctcat aaaggaggcc atccttgaca atgactttat gaagaacttg gagctgtcac
      721 agatccaaga gattgtggat tgtatgtacc cagtggagta cggcaaagac agctgcatca
      781 tcaaagaagg agatgtgggg tcactggtgt atgtcatgga agatggtaag gttgaagtta
      841 caaaagaagg cgtgaagctg tgcacaatgg gtcctggtaa agtgtttgga gagttggcta
      901 tcctttacaa ctgtacccgg acggcgaccg tcaaaactct tgtaaatgtg aaactctggg
      961 ccattgatcg acaatgtttt cagacgataa tgatgaggac aggacttatc aagcataccg
     1021 agtatatgga atttttaaaa agcgttccaa cattccagag ccttcctgaa gagatcctca
     1081 gtaagcttgc tgacgtcctt gaagagaccc actatgaaaa tggggaatat atcatcaggc
     1141 aaggtgcaag aggggacacc ttctttatca tcagtaaagg aaaggttaat gtcactcgtg
     1201 aagactcgcc caatgaagac ccagtctttc ttagaacctt aggaaaagga gattggtttg
     1261 gagagaaagc cttgcagggg gaagatgtga gaacagcgaa tgtaattgcg gcagaagctg
     1321 taacctgcct tgtgatcgac agagactctt tcaaacattt gattggagga ttagatgatg
     1381 tttctaataa agcatatgaa gatgcagaag ctaaggcaaa atatgaagct gaagctgctt
     1441 tcttcgccaa cctgaagaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1501 cgaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc taccactacc
     1561 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1621 cccagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1681 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggtaccggag
     1741 ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1801 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca
     1861 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1921 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1981 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag gagcgcacca
     2041 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2101 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2161 ggcacaagct ggagtacaac ctgtctgatt taacatcatt gacacccttg gagttggagg
     2221 tttcggacga gtagaactgg tccagttaaa aagtgaagaa tccaaaacct ttgcaatgaa
     2281 gattctcaag aaacgccaca attcgcggcc gcactcgaga tgggatccac cggatctaga
     2341 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2401 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2461 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2521 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     2581 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2641 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2701 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2761 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2821 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2881 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2941 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3001 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3061 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3121 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3181 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3241 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3301 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3361 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3421 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3481 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3541 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3601 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3661 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3721 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3781 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3841 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3901 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3961 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4021 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4081 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4141 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4201 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4261 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4321 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4381 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4441 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4501 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4561 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4621 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4681 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4741 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4801 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4861 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4921 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4981 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5041 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5101 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5161 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5221 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5281 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5341 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5401 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5461 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5521 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5581 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5641 gataaccgta ttaccgccat gcat
//
