LOCUS       sequence-266337-        6200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iAChSnFR (non-binding) under hSynapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6200)
  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 6200)
  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.hSynap.iAChSnFR-NULL
FEATURES             Location/Qualifiers
     source          1..6200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   55..184
                     /label=AAV2 ITR
     promoter        266..713
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      742..751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     748..807
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1260..1281
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1384..1405)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2392..2421
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2425..2571
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2587..3175
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2640..2660)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3058..3069)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3178..3194)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3179..3195)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3224..3345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3235..3254)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3289..3308
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3507..3636
                     /label=AAV2 ITR
     primer_bind     complement(3655..3672)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3655..3671)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3664..3686)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3813..4268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3900..3919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4110..4131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4294..4398
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4399..4467,4468..5259)
                     /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(4617..4636)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5430..6018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5919..5938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgcgc
       61 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
      121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
      181 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag
      241 atctctgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg
      301 gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca acccccattc
      361 cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt
      421 gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc
      481 accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc
      541 ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga
      601 ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc
      661 tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt
      721 caagctgcta gcaaggatcc cgccaccatg gagacagaca cactcctgct atgggtactg
      781 ctgctctggg ttccaggttc cactggtgac agatctgcga acgacaccgt agttgtgggc
      841 tcgatcatct ttacagaagg gattatcgtc gcaaacatgg tggcagagat gattgaggcg
      901 catacagacc ttaaggtggt tcgcaaactg aaccttggcg gggtcaacgt taactttgaa
      961 gccattaaac gcggaggtgc gaataatggt attgacattt acgtggagta cactgggcac
     1021 ggtcttgtgg atattctggg gttcccggag ccgaacgtct atatcaccgc cgacaagcag
     1081 aagaacggca tcaaggcgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag
     1141 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1201 aaccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac
     1261 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1321 aagggcggta ccggagggag catgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1381 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc
     1441 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1501 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1561 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1621 caggagcgca ccatcagctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     1681 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1741 ggcaacatcc tggggcacaa gctggagtac aactttccgc cgcccgcgac tactgatcca
     1801 gaaggtgcat acgaaaccgt gaagaaggag tacaaacgta aatggaatat tgtatggctc
     1861 aaaccactgg gattcaacaa tacggcgacg cttaccgtta aagacgaact ggcgaaacag
     1921 tataacctta aaaccttcag tgacttagcg aaaatctcgg ataagctgat tctgggtgca
     1981 acgatgttct ttttagaagg gcccgatggt tacccaggcc tgcaaaaact gtacaatttc
     2041 aaattcaagc acaccaaaag catggacatg ggtattcgct ataccgccat tgataataac
     2101 gaagttcagg taattgatgc ctgggccact gatggcttgc tggtgagcca caaattaaaa
     2161 attctggagg atgataaagc gttcttcccg ccgtattatg ctgcccccat catccgtcag
     2221 gatgtcttag ataagcatcc tgaactgaag gacgtgctga acaaactcgc gaatcaaatt
     2281 tcactggaag aaatgcagaa actgaattac aaggtggacg gtgagggtca ggacccagcg
     2341 aaagtagcta aggagttttt gaaagagaaa ggtttaattc tgcaggtcga cgaacaaaaa
     2401 ctcatctcag aagaggatct gaatgctgtg ggccaggaca cgcaggaggt catcgtggtg
     2461 ccacactcct tgccctttaa ggtggtggtg atctcagcca tcctggccct ggtggtgctc
     2521 accatcatct cccttatcat cctcatcatg ctttggcaga agaagccacg ttagaagctt
     2581 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2641 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2701 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2761 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2821 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2881 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2941 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     3001 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3061 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3121 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     3181 taccgtcgac ctcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag
     3241 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3301 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3361 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     3421 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta
     3481 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     3541 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     3601 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc
     3661 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     3721 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     3781 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     3841 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3901 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3961 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4021 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4081 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4141 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4201 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     4261 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4321 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4381 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4441 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4501 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4561 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4621 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4681 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4741 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4801 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4861 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4921 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4981 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5041 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5101 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5161 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5221 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5281 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5341 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6121 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6181 gtgagcgagg aagcggaaga
//
