LOCUS       sequence-472834-        5061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hChATp is a promoter fragment with activity in striatal cholinergic 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5061)
  AUTHORS   Hunker AC, Wirthlin ME, Gill G, Johansen NJ, Hooper M, Omstead V, 
            Vargas S, Lerma MN, Taskin N, Weed N, Laird WD, Bishaw YM, Bendrick 
            JL, Gore BB, Ben-Simon Y, Opitz-Araya X, Martinez RA, Way SW, 
            Thyagarajan B, Otto S, Sanchez REA, Alexander JR, Amaya A, Amster A,
            Arbuckle J, Ayala A, Baker PM, Barcelli T, Barta S, Bertagnolli D, 
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chong P, Clark M, Colbert K, 
            Daniel S, Dawe T, Departee M, DiValentin P, Donadio NP, Dotson NI, 
            Dwivedi D, Egdorf T, Fliss T, Gary A, Goldy J, Grasso C, Groce EL, 
            Gudsnuk K, Han W, Haradon Z, Hastings S, Helback O, Ho WV, Huang C, 
            Johnson T, Jones DL, Juneau Z, Kenney J, Leibly M, Li S, Liang E, 
            Loeffler H, Lusk NA, Madigan Z, Malloy J, Malone J, McCue R, Melchor
            J, Mich JK, Moosman S, Morin E, Naidoo R, Newman D, Ngo K, Nguyen K,
            Oster AL, Ouellette B, Oyama AA, Pena N, Pham T, Phillips E, Pom C, 
            Potekhina L, Ransford S, Ray PL, Reding M, Rette DF, Reynoldson C, 
            Rimorin C, Sigler AR, Rocha DB, Ronellenfitch K, Ruiz A, Sawyer L, 
            Sevigny JP, Shapovalova NV, Shepard N, Shulga L, Soliman S, Staats 
            B, Taormina MJ, Tieu M, Wang Y, Wilkes J, Wood T, Zhou T, Williford 
            A, Dee N, Mollenkopf T, Ng L, Esposito L, Kalmbach BE, Yao S, Ariza 
            J, Collman F, Mufti S, Smith K, Waters J, Ersing I, Patrick M, Zeng 
            H, Lein ES, Kojima Y, Horwitz G, Owen SF, Levi BP, Daigle TL, Tasic 
            B, Bakken TE, Ting JT
  TITLE     Enhancer AAV toolbox for accessing and perturbing striatal cell 
            types and circuits.
  JOURNAL   Neuron. 2025 May 21;113(10):1507-1524.e17. doi: 
            10.1016/j.neuron.2025.04.035.
  PUBMED    40403704
REFERENCE   2  (bases 1 to 5061)
  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"; doi: 
            "10.1016/j.neuron.2025.04"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1507-1524.e17"
COMMENT     Sequence Label: AiP12701: pAAV-hChATp-SYFP2-WPRE3-BGHpA (Alias...
FEATURES             Location/Qualifiers
     source          1..5061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      425..434
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(431..433,434..436,437..1147)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(476..497)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(737..756)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1084..1105
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1240..1260)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1447..1558
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1676..1816
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1676..1805
                     /label=AAV2 ITR
     rep_origin      1891..2346
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1978..1997)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2188..2209
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2363..2382)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2482..2504
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2542..2560)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2628..2732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2733..2801,2802..3593)
                     /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(2951..2970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3764..4352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4253..4272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4414..4543
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cactgggagg cgtgggaaca agcctgtgga gagtgtccct gcactgtgcc agcaagggct
       61 ttacattctc tgcccagcac tgcaaggcag ccccaggctc ctccatcctt gggcctggtt
      121 catccatctt tctggactct ctgttcctgg caacaaccct gccagtcccg atacatgggc
      181 ggtgcttgcc tctgggtagg ggtggtgttg ggtggggaga ggggtgagga aattagctaa
      241 acctcagctt tagacttgag agaatagagg gtgggctggt ggaaaacatg agtatgtgcc
      301 tgtggaggct taggttcagg gggatctcag agggtagcaa ttgtgaccca cagcctaata
      361 ataacagtct ttgccctctt ggcctggctc ggatccagat ctttcgaagc tagcgctacc
      421 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      481 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      541 tgccacctac ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc
      601 ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga
      661 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      721 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      781 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      841 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa
      901 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt
      961 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1021 cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga
     1081 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1141 gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga tatcataatc aacctctgga
     1201 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     1261 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     1321 ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg ccgcctgcct
     1381 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg ctcgagagat
     1441 cttcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     1501 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     1561 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     1621 aggattggga agacaatagc aggcatgaga tctcacgtgc ggaccgagcg gccgcaggaa
     1681 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     1741 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     1801 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     1861 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     1921 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     1981 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2041 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2101 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2161 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2221 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     2281 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     2341 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     2401 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2461 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2521 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2581 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2701 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2761 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2821 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2881 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2941 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3001 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3061 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3121 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3181 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3241 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3301 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3361 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3421 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3481 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3541 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3601 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3661 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3721 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3781 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3841 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3901 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3961 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4021 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4081 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4141 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4201 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4261 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4321 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4381 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc
     4441 tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     4501 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg cacgcgtccc
     4561 gggagcaggg ggtggggaag gaggagcagt ggctggtggc aagtctgagc caggacatag
     4621 ccgatgacag cctgtgaatg accggccaga ctggggttgg accttcgggg cagagcaggc
     4681 agcccttgga aagccaggct ggtaggagca tgggtgggag gcacaaaaca cagacaaaca
     4741 gacatatata cacatatatc ctctaggcat ctccgaggag tgatggagga gagagttggt
     4801 aggtgggacc acaggaaacc atcccaccca gctctgcctt cttcaagctc agggtccctg
     4861 aagcctgact agtgtctccc gaatggggga ggggggggct ctgggcagag caaaaggagg
     4921 cattggctgc atgtcttagg gcctttacca actgaccttg tccttgctgt cctctcaaaa
     4981 gactgccatg ctccggtgaa tcctcaccag gacgccacct ctttcacttt gcagttgggg
     5041 gcagccccta cgaacccaca g
//
