LOCUS       sequence-374171-        5750 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 5750)
  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 5750)
  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: AAV2-hSyn-G-Flamp1-mut
FEATURES             Location/Qualifiers
     source          1..5750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      579..588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             594..611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(651..659,660..674)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1115..1136
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1242..1263)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1928..1948)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2346..2357)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2495..2971
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2694..2713)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3011..3151
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3011..3140
                     /label=AAV2 ITR
     rep_origin      3226..3681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3313..3332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3523..3544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3698..3717)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3817..3839
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3877..3895)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3963..4067
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4068..4136,4137..4928)
                     /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     complement(4286..4305)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5099..5687
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5588..5607
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5749..5750,1..128)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg
       61 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
      121 gggttcctgc ggccgcacgc gtatgaggcg gggtgggggt gcctacctga cgaccgaccc
      181 cgacccactg gacaagcacc caacccccat tccccaaatt gcgcatcccc tatcagagag
      241 ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac
      301 agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct
      361 gacgtcactc gccggtcccc cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc
      421 cgccgccggc ccagccggac cgcaccacgc gaggcgcgag ataggggggc acgggcgcga
      481 ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg gcagcggagg
      541 agtcgtgtcg tgcctgagag cgcagtcgag aaaccggtgc caccatgggc tcccaccacc
      601 accatcacca tggcatggca agcatgacag gggggcagca gatgggacgg gacctctatg
      661 atgatgacga caaagatccc atgggcttct accaggaggt gagaagaggg gactttgtga
      721 ggaactggca gctagtggca gctgtgcctc tcttccagaa gctgggccca gccgtcctgg
      781 tagaaattgt gagagccctg agagcccgga ccgttcctgc tggggctgtc atctgtcgca
      841 ttggagagcc aggagacagg atgtttttcg ttgtggaagg cagtgtgagt gtggctacca
      901 actggggcaa cgtctatatc acagccgaca agcagaagaa cggcatcaag gcgaacttca
      961 agatccgcca caacgttgag ggcggcggcg tgcagctcgc ctaccactac cagcagaaca
     1021 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc gtgcagtcca
     1081 aactttcgaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1141 ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatgg
     1201 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1261 acgtaaacgg ccacaagttc agcgtgcgtg gcgagggcga gggcgatgcc accaatggca
     1321 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1381 tgaccaccct gacctacggc gtgcagtgct tcgcacgcta ccccgaccac atgaagcagc
     1441 acgacttctt caagtccgcc atgcccgaag gctacatcca ggagcgcacc atcgttttca
     1501 aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1561 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1621 tggagtacaa ccgggtgaat cctgtggagc tgggccctgg agccttcttt ggggagatgg
     1681 ccttgatctc tggagaaccc gaggtggcca ctgtctcagc tgccaccacg gtgagcctgc
     1741 tgtctctgca ctccgctgat ttccaaatgc tgtgcagctc ttccccagag atcgcagaaa
     1801 tcttcaggaa gacagccctg gagagacgag gagcagctgc ctcagcctaa gaattcgata
     1861 tcaagcttat cgatcatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1921 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1981 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2041 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2101 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2161 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2221 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     2281 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2341 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2401 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     2461 cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc
     2521 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     2581 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     2641 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     2701 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     2761 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     2821 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     2881 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     2941 ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc
     3001 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3061 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3121 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3181 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3241 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3301 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3361 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3421 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     3481 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3541 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     3601 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3661 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     3721 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3781 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3841 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3901 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3961 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4021 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4081 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4141 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4201 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4261 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4321 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4381 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4441 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4501 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4561 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4621 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4681 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4741 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4801 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4861 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4921 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4981 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5041 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5101 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5161 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5221 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5281 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5341 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5401 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5461 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5521 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5581 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5641 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5701 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc
//
