LOCUS       sequence-446916-        5339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0869m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal cholinergic neurons (weak).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5339)
  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 5339)
  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: AiP13363 -
            pAAV-AiE0869m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      805..814
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(811..813,814..816,817..1527)
                     /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(1117..1136)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1464..1485
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1537..1593
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1606..1671
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1702..2079
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2166..2186)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2373..2484
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2602..2742
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2602..2731
                     /label=AAV2 ITR
     rep_origin      2817..3272
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2904..2923)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3114..3135
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3289..3308)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3408..3430
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3468..3486)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3554..3658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3659..3727,3728..4519)
                     /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(3877..3896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4690..5278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5179..5198
                     /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 gcgtagaaca ctttctccag gcacagcgga ctcggcacag
      181 gagccaggat tcccggcact tggtctagac ctggggacac aaacagtcat tagtcacagg
      241 ttattataag caggactgtt tcaattgttt taagtgtttc ctcttgcccc tgggaatacc
      301 aggagtgtca aacagcagca acacccacat ttatagctaa gcccatttat gtgaggcaat
      361 attaattgtt gtccactagc actgtttgaa gcagtttcta agcgattccc tgcttcaaat
      421 agccatgttt tggctgggaa gatgctccag tcagcatgtg gtattcactg attgcaagca
      481 cataaaatct gtttacttta ttgctcagct gcgtacaaac acaaggaatg gagtttgatt
      541 cccagaaaaa agccaggtgt gctgggtaat cactggccag actctgatct aatcatcagg
      601 aagccccagg tccctgtgag atcctggttc aaaaaccagg gcggaacagg ctactgagat
      661 aactcagtgg gtagaggcac ttgccaccga gcttgacacc ctgggttcaa tgagctcggg
      721 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      781 ctttcgaagc tagcgctacc ggtcgccacc atggtctcca agggcgagga gctgttcacc
      841 ggggtggtgc ccatcctggt cgagctggac ggcgatgtca acggccacaa gttcagcgtg
      901 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
      961 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1021 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1081 gaaggctacg tccaggagcg caccatcttc ttcaaagacg acggcaacta caagacccgc
     1141 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1201 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1261 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1321 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1381 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1441 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1501 actctcggca tggacgagct gtacaagggc tctggtgcta ccaacttctc actgttgaaa
     1561 caggcagggg atgtagagga gaatccaggg cctggtgcta gtggagacta caaagaccat
     1621 gacggagatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gtccggactc
     1681 agatctggag gctccggagg cccagagcca gcgaagtctg ctcccgcccc gaaaaagggc
     1741 tccaagaagg cggtgactaa ggcgcagaag aaaggcggca agaagcgcaa gcgcagccgc
     1801 aaggagagct attccatcta tgtgtacaag gttctgaagc aggtccaccc tgacaccggc
     1861 atttcgtcca aggccatggg catcatgaat tcgtttgtga acgacatttt cgagcgcatc
     1921 gcaggagagg cttcccgcct ggcgcattac aacaagcgct cgaccatcac ctcccgggag
     1981 atccagacgg ccgtgcgcct gctgctgcct ggggagttgg ccaagcacgc cgtgtccgag
     2041 ggtactaagg ccatcaccaa gtacaccagc gctaagtaat gaggcgcgcc gcggccgcga
     2101 attcgatatc ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2161 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2221 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     2281 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2341 actgacaatt ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt
     2401 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2461 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2521 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc
     2581 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2641 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2701 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     2761 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     2821 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2881 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2941 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3001 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     3061 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3121 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     3181 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3241 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     3301 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3361 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3421 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3481 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3541 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3601 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3661 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3721 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3781 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3841 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3901 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3961 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4021 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4081 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4141 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4201 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4261 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4321 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4381 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4441 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4501 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4561 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4621 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4681 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4741 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4801 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4861 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4921 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4981 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5041 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5101 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5161 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5221 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5281 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
