LOCUS       sequence-266350-        6885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLEXed iAChSnFR (yellow version) under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6885)
  AUTHORS   Borden PM, Zhang P, Shivange AV, Marvin JS, Cichon J, Dan C, 
            Podgorski K, Figueiredo A, Novak O, Tanimoto M, Shigetomi E, Lobas 
            MA, Kim H, Zhu PK, Zhang Y, Zheng WS, Fan C, Wang G, Xiang B, Gan L,
            Zhang G, Guo K, Lin L, Cai Y, Yee AG, Aggarwal A, Ford CP, Rees DC, 
            Dietrich D, Khakh BS, Dittman JS, Gan W, Koyama M, Jayaraman V, 
            Cheer JF, Lester HA, Zhu JJ, Looger LL
  TITLE     A fast genetically encoded fluorescent sensor for faithful in vivo 
            acetylcholine detection in mice, fish, worms and flies
  JOURNAL   bioRxiv 2020
REFERENCE   2  (bases 1 to 6885)
  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: "bioRxiv 
            2020"
COMMENT     Sequence Label: pAAV.CAG-FLEX.iAChSnFR-Venus
FEATURES             Location/Qualifiers
     source          1..6885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1133..1166
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1237..1270
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     sig_peptide     1299..1358
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1811..1832
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1935..1956)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2943..2972
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2976..3122
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(3158..3191)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3254..3287)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(3325..3344)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3406..3428)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3708..3985)
                     /label=chicken beta-actin promoter
     repeat_region   4298..4427
                     /label=AAV2 ITR
     rep_origin      4499..4954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4586..4605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4796..4817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4980..5084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5085..5153,5154..5945)
                     /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(5303..5322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6116..6704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6605..6624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct aattcgaacc tcttcgaggg acataacttc
     1141 gtataatgta tgctatacga agttatgggt cgatggtgag atctggacta gagggtcgat
     1201 ggtgatgctt ggatcaaaat aggaagaatc ccaatgataa cttcgtataa agtatcctat
     1261 acgaagttat ggatccgaat tcactagtct cgagcaaaat ggagacagac acactcctgc
     1321 tatgggtact gctgctctgg gttccaggtt ccactggtga cagatctgcg aacgacaccg
     1381 tagttgtggg ctcgatcatc tttacagaag ggattatcgt cgcaaacatg gtggcagaga
     1441 tgattgaggc gcatacagac cttaaggtgg ttcgcaaact gaaccttggc ggggtcaacg
     1501 ttaactttga agccattaaa cgcggaggtg cgaataatgg tattgacatt tacgtggagt
     1561 acactgggca cggtcttgtg gatattctgg ggttcccgga gccgaacgtc tatatcaccg
     1621 ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac gtggaggacg
     1681 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1741 tgctgcccga caaccactac ctgagctatc agtccgtgct gagcaaagac cccaacgaga
     1801 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatga
     1861 acgagctgta caagggcggt accggaggga gcatgagcaa gggcgaggag ctgttcaccg
     1921 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc
     1981 gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagctc ctgtgcacca
     2041 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt
     2101 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2161 aaggctacgt ccaggagcgc accatcagct tcaaggacga cggcacctac aagacccgcg
     2221 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2281 tcaaggagga cggcaacatc ctggggcaca agctggagta caactttccg ccgcccgcga
     2341 ctactgatcc agaaggtgca tacgaaaccg tgaagaagga gtacaaacgt aaatggaata
     2401 ttgtatggct caaaccactg ggattcaaca atacgtatac gcttaccgtt aaagacgaac
     2461 tggcgaaaca gtataacctt aaaaccttca gtgacttagc gaaaatctcg gataagctga
     2521 ttctgggtgc aacgatgttc tttttagaag ggcccgatgg ttacccaggc ctgcaaaaac
     2581 tgtacaattt caaattcaag cacaccaaaa gcatggacat gggtattcgc tataccgcca
     2641 ttgataataa cgaagttcag gtaattgatg cctgggccac tgatggcttg ctggtgagcc
     2701 acaaattaaa aattctggag gatgataaag cgttcttccc gccgtattat gctgccccca
     2761 tcatccgtca ggatgtctta gataagcatc ctgaactgaa ggacgtgctg aacaaactcg
     2821 cgaatcaaat ttcactggaa gaaatgcaga aactgaatta caaggtggac ggtgagggtc
     2881 aggacccagc gaaagtagct aaggagtttt tgaaagagaa aggtttaatt ctgcaggtcg
     2941 acgaacaaaa actcatctca gaagaggatc tgaatgctgt gggccaggac acgcaggagg
     3001 tcatcgtggt gccacactcc ttgcccttta aggtggtggt gatctcagcc atcctggccc
     3061 tggtggtgct caccatcatc tcccttatca tcctcatcat gctttggcag aagaagccac
     3121 gttaggatat caccggtggc gcgccaagct tgctagtata acttcgtata gcatacatta
     3181 tacgaagtta tgggtcgatg gtgatgcttg gcaattcggg tcgatggtga agcattggtc
     3241 ttcctatttt gatataactt cgtataggat actttatacg aagttatggg gatgatccaa
     3301 ttctttgcca aaatgatgag acagcacaat aaccagcacg ttgcccagga gctgtaggaa
     3361 aaagaagaag gcatgaacat ggttagcaga ggctctagag ccgccggtca cacgccagaa
     3421 gccgaacccc gccctgcccc gtcccccccg aaggcagccg tccccccgcg gacagccccg
     3481 agccgctccc cccgcacaaa gggccctccc ggagcccctc aaggctttca cgcagccaca
     3541 gaaaagaaac aagccgtcat taaaccaagc gctaattaca gcccggagga gaagggccgt
     3601 cccgcccgct cacctgtggg agtaacgcgg tcagtcagag ccggggcggg cggcgcgagg
     3661 cggcggcgga gcggggcacg gggcgaaggc agcgtcgcag cgactcccgc ccgccgcgcg
     3721 cttcgctttt tatagggccg ccgccgccgc cgcctcgcca taaaaggaaa ctttcggagc
     3781 gcgccgctct gattggctgc cgccgcacct ctccgcctcg ccccgccccg cccctcgccc
     3841 cgccccgccc cgcctggcgc gcgccccccc cccccccccg cccccatcgc tgcacaaaat
     3901 aattaaaaaa taaataaata caaaattggg ggtggggagg ggggggagat ggggagagtg
     3961 aagcagaacg tggggctcac ctcgaccatg gtaatagcga tgactaatac gtagatgtac
     4021 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     4081 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     4141 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     4201 actatgggaa catacgtcat tattgacgtc aatggtacga tcttcctaga gcatggctac
     4261 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     4321 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     4381 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg gcgtaatagc
     4441 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     4501 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4561 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4621 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4681 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4741 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4801 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4861 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4921 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     4981 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5041 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5101 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5161 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5221 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5281 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5341 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5401 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5461 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5521 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5581 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5641 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5701 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5761 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5821 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5881 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5941 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6001 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6061 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6121 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6181 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6241 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6481 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6661 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6721 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6781 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6841 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
