LOCUS       sequence-271681-        6212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iAChSnFR under hSynapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6212)
  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 6212)
  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
FEATURES             Location/Qualifiers
     source          1..6212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   114..243
                     /label=AAV2 ITR
     promoter        325..772
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      801..810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     807..866
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1319..1340
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1443..1464)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2451..2480
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2484..2630
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2646..3234
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2699..2719)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3117..3128)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3237..3253)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3238..3254)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3283..3404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3294..3313)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3348..3367
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3566..3695
                     /label=AAV2 ITR
     rep_origin      3767..4222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3854..3873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4064..4085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4248..4352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4353..4421,4422..5213)
                     /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(4571..4590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5384..5972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5873..5892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
       61 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg
      121 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
      181 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      241 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
      301 tctctgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga tgaggcgggg
      361 tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc
      421 ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg
      481 cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca
      541 ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct
      601 tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag
      661 gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct
      721 gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc agtcgaattc
      781 aagctgctag caaggatccc gccaccatgg agacagacac actcctgcta tgggtactgc
      841 tgctctgggt tccaggttcc actggtgaca gatctgcgaa cgacaccgta gttgtgggct
      901 cgatcatctt tacagaaggg attatcgtcg caaacatggt ggcagagatg attgaggcgc
      961 atacagacct taaggtggtt cgcaaactga accttggcgg ggtcaacgtt aactttgaag
     1021 ccattaaacg cggaggtgcg aataatggta ttgacattta cgtggagtac actgggcacg
     1081 gtcttgtgga tattctgggg ttcccggagc cgaacgtcta tatcaccgcc gacaagcaga
     1141 agaacggcat caaggcgaac ttcaagatcc gccacaacgt ggaggacggc agcgtgcagc
     1201 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1261 accactacct gagcacccag tccgtgctga gcaaagaccc caacgagaag cgcgatcaca
     1321 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1381 agggcggtac cggagggagc atgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1441 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgcgc ggcgagggcg
     1501 agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1561 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     1621 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1681 aggagcgcac catcagcttc aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt
     1741 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1801 gcaacatcct ggggcacaag ctggagtaca actttccgcc gcccgcgact actgatccag
     1861 aaggtgcata cgaaaccgtg aagaaggagt acaaacgtaa atggaatatt gtatggctca
     1921 aaccactggg attcaacaat acgtatacgc ttaccgttaa agacgaactg gcgaaacagt
     1981 ataaccttaa aaccttcagt gacttagcga aaatctcgga taagctgatt ctgggtgcaa
     2041 cgatgttctt tttagaaggg cccgatggtt acccaggcct gcaaaaactg tacaatttca
     2101 aattcaagca caccaaaagc atggacatgg gtattcgcta taccgccatt gataataacg
     2161 aagttcaggt aattgatgcc tgggccactg atggcttgct ggtgagccac aaattaaaaa
     2221 ttctggagga tgataaagcg ttcttcccgc cgtattatgc tgcccccatc atccgtcagg
     2281 atgtcttaga taagcatcct gaactgaagg acgtgctgaa caaactcgcg aatcaaattt
     2341 cactggaaga aatgcagaaa ctgaattaca aggtggacgg tgagggtcag gacccagcga
     2401 aagtagctaa ggagtttttg aaagagaaag gtttaattct gcaggtcgac gaacaaaaac
     2461 tcatctcaga agaggatctg aatgctgtgg gccaggacac gcaggaggtc atcgtggtgc
     2521 cacactcctt gccctttaag gtggtggtga tctcagccat cctggccctg gtggtgctca
     2581 ccatcatctc ccttatcatc ctcatcatgc tttggcagaa gaagccacgt tagaagctta
     2641 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2701 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2761 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2821 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2881 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2941 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3001 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3061 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3121 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3181 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3241 accgtcgacc tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt
     3301 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3361 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3421 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     3481 tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag
     3541 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     3601 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3661 gcccgggcgg cctcagtgag cgagcgagcg cgcagctggc gtaatagcga agaggcccgc
     3721 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     3781 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3841 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3901 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3961 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4021 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4081 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4141 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4201 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4261 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4321 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4381 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4441 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4501 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4561 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4621 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4681 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4741 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4801 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4861 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4921 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4981 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5041 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5101 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5161 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5221 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5281 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5341 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5401 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5461 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5521 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5581 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5641 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5701 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5761 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5821 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5881 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5941 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6001 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6061 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6121 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     6181 ccgcgcgttg gccgattcat taatgcagct gg
//
