LOCUS       sequence-446773-        5451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1417m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal Pvalb+/Pthlh+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5451)
  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 5451)
  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: AiP15041 -
            pAAV-AiE1417m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5451
                     /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      804..813
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(810..812,813..815,816..1526)
                     /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(1116..1135)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1463..1484
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1536..1592
                     /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             1605..1670
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1701..2078
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2165..2185)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2372..2483
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2601..2741
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2601..2730
                     /label=AAV2 ITR
     misc_feature    2748..2775
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2776..2803
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2872..3327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2959..2978)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3169..3190
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3344..3363)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3463..3485
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3523..3541)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3609..3713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3714..3782,3783..4574)
                     /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(3932..3951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4745..5333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5234..5253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5390..5417
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5418..5445
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtgca ggaatagaaa ccctgactaa gacactcact
      181 ttcccctctt tgttatcaaa gctgctgcca gccactgggc tcctctcctg tgaccgatct
      241 atagttttca agtgttttct ttgagcaggc ttttgtgtgt gtatagtgtg tgatcagaaa
      301 gattatctta ttccagagga catgtgtgtg cttaacctga tatcttttcc caggaagcta
      361 gactcctctg cctagggatg ttcaacaccc ctcctgaggc ccggtccatg ctccccgctt
      421 taatgtgttc agaaaccctc agctcagcac caggcactct ccaggcactg ctcttatcct
      481 cacttcctga gattgagcac agtgaacttg ctgagccagc agcctgggga tgctctgctc
      541 tctctggaag tgcttttggg cttaaagaat ggtttctgca gctttgtgag cagaagccac
      601 acagcctttt cttctcccct ggaccacagg atttactggc acgcacaagg acggtcacaa
      661 aggtcgccca cccctgcccc caagctttag tgatttatat tcccaagtca gagctcgggc
      721 tgggcataaa agtcagggca gagccatcta ttgcttacat ttgcttctgg gatccagatc
      781 tttcgaagct agcgctaccg gtcgccacca tggtctccaa gggcgaggag ctgttcaccg
      841 gggtggtgcc catcctggtc gagctggacg gcgatgtcaa cggccacaag ttcagcgtgt
      901 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca
      961 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt
     1021 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1081 aaggctacgt ccaggagcgc accatcttct tcaaagacga cggcaactac aagacccgcg
     1141 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1201 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1261 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     1321 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1381 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag
     1441 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1501 ctctcggcat ggacgagctg tacaagggct ctggtgctac caacttctca ctgttgaaac
     1561 aggcagggga tgtagaggag aatccagggc ctggtgctag tggagactac aaagaccatg
     1621 acggagatta taaagatcat gacatcgatt acaaggatga cgatgacaag tccggactca
     1681 gatctggagg ctccggaggc ccagagccag cgaagtctgc tcccgccccg aaaaagggct
     1741 ccaagaaggc ggtgactaag gcgcagaaga aaggcggcaa gaagcgcaag cgcagccgca
     1801 aggagagcta ttccatctat gtgtataagg ttctgaagca ggtccaccct gacaccggca
     1861 tttcgtccaa ggccatgggc atcatgaact cgtttgtgaa cgacattttc gagcgcatcg
     1921 caggtgaggc ttcccgcctg gcgcattaca acaagcgctc gaccatcacc tccagggaga
     1981 tccagacggc cgtgcgcctg ctgctgcctg gggagttggc caagcacgcc gtgtccgagg
     2041 gtactaaggc catcaccaag tacaccagcg ctaagtaatg aggcgcgccg cggccgcgaa
     2101 ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2161 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2221 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     2281 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2341 ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     2401 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2461 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2521 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     2581 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2641 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2701 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctat cagcacacaa
     2761 ttgcccatta tacgcgcgta taatggacta ttgtgtgctg atatgatgcg gtattttctc
     2821 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     2881 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2941 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3001 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3061 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3121 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3181 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     3241 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3301 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3361 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3421 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3481 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3541 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcat atcagcacac
     5401 aattgcccat tatacgcgcg tataatggac tattgtgtgc tgataacatg t
//
