LOCUS       sequence-271580-        6181 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses FLEXed iAChSnFR (yellow version) under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6181)
  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 6181)
  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..6181
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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)."
     primer_bind     1812..1833
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1936..1957)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2944..2973
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2977..3123
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(3159..3192)
                     /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(3255..3288)
                     /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(3326..3345)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3407..3429)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     repeat_region   3594..3723
                     /label=AAV2 ITR
     rep_origin      3795..4250
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3882..3901)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4092..4113
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4276..4380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4381..4449,4450..5241)
                     /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(4599..4618)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5412..6000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5901..5920
                     /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 ggagacaaga cacactcctg
     1321 ctatgggtac tgctgctctg ggttccaggt tccactggtg acagatctgc gaacgacacc
     1381 gtagttgtgg gctcgatcat ctttacagaa gggattatcg tcgcaaacat ggtggcagag
     1441 atgattgagg cgcatacaga ccttaaggtg gttcgcaaac tgaaccttgg cggggtcaac
     1501 gttaactttg aagccattaa acgcggaggt gcgaataatg gtattgacat ttacgtggag
     1561 tacactgggc acggtcttgt ggatattctg gggttcccgg agccgaacgt ctatatcacc
     1621 gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa cgtggaggac
     1681 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1741 ctgctgcccg acaaccacta cctgagctat cagtccgtgc tgagcaaaga ccccaacgag
     1801 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1861 aacgagctgt acaagggcgg taccggaggg agcatgagca agggcgagga gctgttcacc
     1921 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1981 cgcggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagct cctgtgcacc
     2041 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     2101 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     2161 gaaggctacg tccaggagcg caccatcagc ttcaaggacg acggcaccta caagacccgc
     2221 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2281 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactttcc gccgcccgcg
     2341 actactgatc cagaaggtgc atacgaaacc gtgaagaagg agtacaaacg taaatggaat
     2401 attgtatggc tcaaaccact gggattcaac aatacgtata cgcttaccgt taaagacgaa
     2461 ctggcgaaac agtataacct taaaaccttc agtgacttag cgaaaatctc ggataagctg
     2521 attctgggtg caacgatgtt ctttttagaa gggcccgatg gttacccagg cctgcaaaaa
     2581 ctgtacaatt tcaaattcaa gcacaccaaa agcatggaca tgggtattcg ctataccgcc
     2641 attgataata acgaagttca ggtaattgat gcctgggcca ctgatggctt gctggtgagc
     2701 cacaaattaa aaattctgga ggatgataaa gcgttcttcc cgccgtatta tgctgccccc
     2761 atcatccgtc aggatgtctt agataagcat cctgaactga aggacgtgct gaacaaactc
     2821 gcgaatcaaa tttcactgga agaaatgcag aaactgaatt acaaggtgga cggtgagggt
     2881 caggacccag cgaaagtagc taaggagttt ttgaaagaga aaggtttaat tctgcaggtc
     2941 gacgaacaaa aactcatctc agaagaggat ctgaatgctg tgggccagga cacgcaggag
     3001 gtcatcgtgg tgccacactc cttgcccttt aaggtggtgg tgatctcagc catcctggcc
     3061 ctggtggtgc tcaccatcat ctcccttatc atcctcatca tgctttggca gaagaagcca
     3121 cgttaggata tcaccggtgg cgcgccaagc ttgctagtat aacttcgtat agcatacatt
     3181 atacgaagtt atgggtcgat ggtgatgctt ggcaattcgg gtcgatggtg aagcattggt
     3241 cttcctattt tgatataact tcgtatagga tactttatac gaagttatgg ggatgatcca
     3301 attctttgcc aaaatgatga gacagcacaa taaccagcac gttgcccagg agctgtagga
     3361 aaaagaagaa ggcatgaaca tggttagcag aggctctaga gccgccggtc acacgccaga
     3421 agccgaaccc cgccctgccc cgtccccccc gaaggcagcc gtccccccgc ggacagcccc
     3481 gagccgctcc cccgcgttac tatgggaaca tacgtcatta ttgacgtcaa tggtactctt
     3541 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     3601 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3661 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3721 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     3781 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3841 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3901 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3961 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4021 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4081 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4141 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4201 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     4261 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4321 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4381 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4441 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4501 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4561 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4621 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4681 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4741 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4801 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4861 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4921 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4981 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5041 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5101 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5161 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5221 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5281 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5341 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5401 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5461 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5521 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5581 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5641 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5701 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5761 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5821 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5881 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5941 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6001 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6061 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6181 g
//
