LOCUS       sequence-374170-        8018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  control probe of G-Flamp1, insensitive to cAMP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8018)
  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 8018)
  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-mut
FEATURES             Location/Qualifiers
     source          1..8018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        132..435
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1121..2137
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2061..2083
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2145..2164
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2239..2248
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2254..2271
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2275..2307
                     /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(2311..2319,2320..2334)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     2775..2796
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2902..2923)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3562..3591
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3607..3624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3647..3664)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3653..3877
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3923..4351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4010..4029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4220..4241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4360..4380)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4365..4694
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4545..4680
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4607..4626
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4761..5555
                     /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(4815..4834)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5425..5444
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5731..5852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5768..5787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5822..5841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5901..5917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5901..5917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5914..5936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5925..5941
                     /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(5949..5955,5956..5973,5974..5979))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5994..6015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6132..6149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6303..6891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6383..6402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7062..7853,7854..7922))
                     /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     7685..7704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7923..8018,1..9))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaatt
       61 ctgcaggaat ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      121 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      181 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      241 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      301 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      361 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      421 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
      481 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
      541 gcgggggggg ggggggggss sscscsrggg gggggggggg gggggggggg gggggggggg
      601 gggggggggg gggggggggg gggggscmmm mmrrgggggg ssscccssrr rkttyytttt
      661 ttkkssgggg gggggggggg ggggscccyw waaaaargrr rsssssgggg ggggggggnn
      721 nnnngsmrss scssstgnnn nnnngkgwaa wgssccscck ggmwttwtsc ccrktmcwwk
      781 vccttwtggg vytttccyyy ttggsmktwc myythcktwt twkyymycsc ywttyccmkg
      841 gkcgrggkgr sccccccbtt ytbctycccc cccccccycc cccccccccc ccccccccca
      901 attttktwtt tttttttttt ttwattwttt tgggsggggg gggggggggg gggggggggg
      961 gggncgcgcg ccrggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
     1021 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
     1081 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
     1141 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     1201 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
     1261 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
     1321 gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
     1381 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
     1441 ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
     1501 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
     1561 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga
     1621 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc
     1681 gggcgggggg tggcggcagg kgggggtgcc gggcggggcg gggccgccyc gggcsggggr
     1741 gggckcgggg grggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
     1801 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
     1861 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
     1921 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
     1981 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
     2041 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     2101 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     2161 tgtgctgtct catcattttg gcaaagaatt gatttgatac cgcgggcccg ggatcagctt
     2221 ggtaccgagc tcggatccgc caccatgggt tctcatcatc atcatcatca tggtatggct
     2281 agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga taaggatccg
     2341 atgggcttct atcaagaagt ccgtcgcggg gatttcgtcc gcaattggca attggtcgcc
     2401 gccgtgccgt tgtttcagaa gctcggcccg gccgtgctgg tcgagatcgt gcgcgcctta
     2461 agagcccgca cggtgccggc gggcgccgtg atctgccgca ttggcgagcc cggcgatcgg
     2521 atgttcttcg tcgtggaggg gagcgtcagc gtcgcgacga attgggggaa cgtctatatc
     2581 acagccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca caacgttgag
     2641 ggcggcggcg tgcagctcgc ctaccactac cagcagaaca cccccatcgg cgacggcccc
     2701 gtgctgctgc ccgacaacca ctacctgagc gtgcagtcca aactttcgaa agaccccaac
     2761 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2821 atggacgagc tgtacaaggg cggtaccgga gggagcatgg tgagcaaggg cgaggagctg
     2881 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2941 agcgtgcgtg gcgagggcga gggcgatgcc accaatggca agctgaccct gaagttcatc
     3001 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     3061 gtgcagtgct tcgcacgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3121 atgcccgaag gctacatcca ggagcgcacc atcgttttca aggacgacgg cacctacaag
     3181 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3241 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ccgcgtgaat
     3301 ccggtggagc ttggccctgg cgccttcttc ggcgagatgg cgctgatcag cggcgaaccg
     3361 gaggttgcga ccgtcagcgc cgcaacgacg gtctcactcc tgtcgctgca ttcggcggat
     3421 ttccagatgt tgtgcagcag cagcccggag atcgcggaaa tcttccgcaa gaccgcgctc
     3481 gagcgtcgcg gcgctgcggc gagcgcttaa tgaattctgc agatatccag cacagtggcg
     3541 gccgctcgag gtcacccatt cgaacaaaaa ctcatctcag aagaggatct gaatatgcat
     3601 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
     3661 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3721 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3781 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3841 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3901 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3961 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4021 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4081 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4141 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4201 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4261 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4321 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4381 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4441 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4501 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4561 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4621 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4681 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4741 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4801 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4861 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4921 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4981 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5041 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5101 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5161 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5221 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5281 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5341 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5401 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5461 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5521 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     5581 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5641 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5701 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5821 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5881 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     6001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     6061 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6721 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7201 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7981 tacatatttg aatgtattta gaaaaataaa caaatagg
//
