LOCUS       sequence-271715-        6414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iAChSnFR (non-binding variant) under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6414)
  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 6414)
  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.iAChSnFR-NULL
FEATURES             Location/Qualifiers
     source          1..6414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        223..526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        528..805
                     /label=chicken beta-actin promoter
     primer_bind     1061..1080
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     sig_peptide     1135..1194
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1647..1668
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1771..1792)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2779..2808
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2812..2958
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     polyA_signal    2963..3097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2987..3006)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3041..3060
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3183..3200)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3189..3396
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3426..3566
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3426..3555
                     /label=AAV2 ITR
     rep_origin      3652..4107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3739..3758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3949..3970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4174..4193)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4293..4315
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4353..4371)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4439..4543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4544..4612,4613..5404)
                     /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(4762..4781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5575..6163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6064..6083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtagatct gaattcggta cctagttatt aatagtaatc
      181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      481 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      541 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      601 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccrgg
      661 gggggsgggg gggggsgggg ggsggggsgg ggsgaggcgg agaggtgcgg cggcagccaa
      721 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      781 taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg
      841 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
      901 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
      961 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagctag agcctctgct
     1021 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1081 ctgtctcatc attttggcaa agaattcctc gaagatccga aggggttcaa gcttatggag
     1141 acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac tggtgacaga
     1201 tctgcgaacg acaccgtagt tgtgggctcg atcatcttta cagaagggat tatcgtcgca
     1261 aacatggtgg cagagatgat tgaggcgcat acagacctta aggtggttcg caaactgaac
     1321 cttggcgggg tcaacgttaa ctttgaagcc attaaacgcg gaggtgcgaa taatggtatt
     1381 gacatttacg tggagtacac tgggcacggt cttgtggata ttctggggtt cccggagccg
     1441 aacgtctata tcaccgccga caagcagaag aacggcatca aggcgaactt caagatccgc
     1501 cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1561 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgtgctgagc
     1621 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1681 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat gagcaagggc
     1741 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1801 cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
     1861 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1921 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     1981 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc
     2041 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2101 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2161 tttccgccgc ccgcgactac tgatccagaa ggtgcatacg aaaccgtgaa gaaggagtac
     2221 aaacgtaaat ggaatattgt atggctcaaa ccactgggat tcaacaatac ggcgacgctt
     2281 accgttaaag acgaactggc gaaacagtat aaccttaaaa ccttcagtga cttagcgaaa
     2341 atctcggata agctgattct gggtgcaacg atgttctttt tagaagggcc cgatggttac
     2401 ccaggcctgc aaaaactgta caatttcaaa ttcaagcaca ccaaaagcat ggacatgggt
     2461 attcgctata ccgccattga taataacgaa gttcaggtaa ttgatgcctg ggccactgat
     2521 ggcttgctgg tgagccacaa attaaaaatt ctggaggatg ataaagcgtt cttcccgccg
     2581 tattatgctg cccccatcat ccgtcaggat gtcttagata agcatcctga actgaaggac
     2641 gtgctgaaca aactcgcgaa tcaaatttca ctggaagaaa tgcagaaact gaattacaag
     2701 gtggacggtg agggtcagga cccagcgaaa gtagctaagg agtttttgaa agagaaaggt
     2761 ttaattctgc aggtcgacga acaaaaactc atctcagaag aggatctgaa tgctgtgggc
     2821 caggacacgc aggaggtcat cgtggtgcca cactccttgc cctttaaggt ggtggtgatc
     2881 tcagccatcc tggccctggt ggtgctcacc atcatctccc ttatcatcct catcatgctt
     2941 tggcagaaga agccacgtta gggatccaga catgataaga tacattgatg agtttggaca
     3001 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3061 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3121 tatgtttcag gttcaggggg aggtgtggga ggttttttag tcgactagag ctcgctgatc
     3181 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3241 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3301 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3361 ggaggattgg gaagacaata gcaggcatgc tggggagaga tctcacgtgc ggaccgagcg
     3421 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     3481 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     3541 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcca gctggcgtaa tagcgaagag
     3601 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     3661 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3721 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3781 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3841 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3901 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3961 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4021 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4081 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     4141 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4201 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4261 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4321 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4381 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4441 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4501 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4561 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4621 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4681 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4741 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4801 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4861 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4921 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4981 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5041 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5101 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5161 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5221 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5281 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5341 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5401 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5461 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5521 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5581 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5641 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5701 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5761 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5821 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5881 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5941 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6001 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6061 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6121 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6181 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6241 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6301 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6361 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggggctcac atgt
//
