LOCUS       sequence-271575-        6882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLEXed iAChSnFR (non-binding yellow version) under CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6882)
  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 6882)
  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-NULL
FEATURES             Location/Qualifiers
     source          1..6882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        181..458
                     /label=chicken beta-actin promoter
     primer_bind     737..759
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     821..840
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    878..911
                     /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)."
     protein_bind    974..1007
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1043..1189)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1193..1222)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     2209..2230
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2333..2354)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2807..2866)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    complement(2895..2928)
                     /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)."
     protein_bind    complement(2999..3032)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3066..3654
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3119..3139)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3537..3548)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3697..3818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3708..3727)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3762..3781
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3980..4109
                     /label=AAV2 ITR
     rep_origin      complement(4346..4934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4426..4445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5105..5896,5897..5965))
                     /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     5728..5747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5966..6070)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6096..6551)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6233..6254)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6445..6464
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6623..6752
                     /label=AAV2 ITR
ORIGIN
        1 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
       61 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      121 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      181 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
      241 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      301 ggcgcgcgcc rggggggggg gggsggggsg rggggsgggg sggggsgrgg cggagaggtg
      361 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
      421 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
      481 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      541 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
      601 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
      661 gccctttgtg cggggggagc ggctcggggc tgtccgcggg gggacggctg ccttcggggg
      721 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
      781 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
      841 ctgtctcatc attttggcaa agaattggat catccccata acttcgtata aagtatccta
      901 tacgaagtta tatcaaaata ggaagaccaa tgcttcacca tcgacccgaa ttgccaagca
      961 tcaccatcga cccataactt cgtataatgt atgctatacg aagttatact agcaagcttg
     1021 gcgcgccacc ggtgatatcc taacgtggct tcttctgcca aagcatgatg aggatgataa
     1081 gggagatgat ggtgagcacc accagggcca ggatggctga gatcaccacc accttaaagg
     1141 gcaaggagtg tggcaccacg atgacctcct gcgtgtcctg gcccacagca ttcagatcct
     1201 cttctgagat gagtttttgt tcgtcgacct gcagaattaa acctttctct ttcaaaaact
     1261 ccttagctac tttcgctggg tcctgaccct caccgtccac cttgtaattc agtttctgca
     1321 tttcttccag tgaaatttga ttcgcgagtt tgttcagcac gtccttcagt tcaggatgct
     1381 tatctaagac atcctgacgg atgatggggg cagcataata cggcgggaag aacgctttat
     1441 catcctccag aatttttaat ttgtggctca ccagcaagcc atcagtggcc caggcatcaa
     1501 ttacctgaac ttcgttatta tcaatggcgg tatagcgaat acccatgtcc atgcttttgg
     1561 tgtgcttgaa tttgaaattg tacagttttt gcaggcctgg gtaaccatcg ggcccttcta
     1621 aaaagaacat cgttgcaccc agaatcagct tatccgagat tttcgctaag tcactgaagg
     1681 ttttaaggtt atactgtttc gccagttcgt ctttaacggt aagcgtcgcc gtattgttga
     1741 atcccagtgg tttgagccat acaatattcc atttacgttt gtactccttc ttcacggttt
     1801 cgtatgcacc ttctggatca gtagtcgcgg gcggcggaaa gttgtactcc agcttgtgcc
     1861 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     1921 tgtcgccctc gaacttcacc tcggcgcggg tcttgtaggt gccgtcgtcc ttgaagctga
     1981 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     2041 tgtggtcggg gtagcgggcg aagcactgca cgccgtagcc cagggtggtc acgagggtgg
     2101 gccagggcac gggcagcttg ccggtggtgc acaggagctt cagggtcagc ttgccgttgg
     2161 tggcatcgcc ctcgccctcg ccgcgcacgc tgaacttgtg gccgtttacg tcgccgtcca
     2221 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc atgctccctc
     2281 cggtaccgcc cttgtacagc tcgttcatgc cgagagtgat cccggcggcg gtcacgaact
     2341 ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagcacg gactgatagc
     2401 tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt
     2461 agtggtcggc gagctgcacg ctgccgtcct ccacgttgtg gcggatcttg aagttcgcct
     2521 tgatgccgtt cttctgcttg tcggcggtga tatagacgtt cggctccggg aaccccagaa
     2581 tatccacaag accgtgccca gtgtactcca cgtaaatgtc aataccatta ttcgcacctc
     2641 cgcgtttaat ggcttcaaag ttaacgttga ccccgccaag gttcagtttg cgaaccacct
     2701 taaggtctgt atgcgcctca atcatctctg ccaccatgtt tgcgacgata atcccttctg
     2761 taaagatgat cgagcccaca actacggtgt cgttcgcaga tctgtcacca gtggaacctg
     2821 gaacccagag cagcagtacc catagcagga gtgtgtctgt ctccattttg ctcgagacta
     2881 gtgaattcgg atccataact tcgtatagga tactttatac gaagttatca ttgggattct
     2941 tcctattttg atccaagcat caccatcgac cctctagtcc agatctcacc atcgacccat
     3001 aacttcgtat agcatacatt atacgaagtt atgtccctcg aagaggttcg aattagctta
     3061 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3121 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3181 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3241 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3301 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3361 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3421 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3481 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3541 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3601 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3661 accgtcgacc cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     3721 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     3781 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     3841 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     3901 aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc
     3961 gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4021 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4081 gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga
     4141 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4201 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4261 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4321 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4381 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4441 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4501 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4561 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4621 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4681 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4741 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4801 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4861 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4921 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4981 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5041 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5101 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5161 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5221 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5281 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5341 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5401 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5461 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5521 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5581 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5641 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5701 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5761 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5821 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5881 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5941 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6001 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6061 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     6121 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     6181 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     6241 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     6301 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     6361 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     6421 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     6481 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     6541 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6601 ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
     6661 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
     6721 gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta
     6781 cttatctacg tagccatgct ctaggaagag taccattgac gtcaataatg acgtatgttc
     6841 ccatagtaac gccaataggg actttccatt gacgtcaatg gg
//
