LOCUS       sequence-446775-        5441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1418m 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 5441)
  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 5441)
  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: AiP15042 -
            pAAV-AiE1418m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5441
                     /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      794..803
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(800..802,803..805,806..1516)
                     /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(1106..1125)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1453..1474
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1526..1582
                     /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             1595..1660
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1691..2068
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2155..2175)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2362..2473
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2591..2731
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2591..2720
                     /label=AAV2 ITR
     misc_feature    2738..2765
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2766..2793
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2862..3317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2949..2968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3159..3180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3334..3353)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3453..3475
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3513..3531)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3599..3703
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3704..3772,3773..4564)
                     /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(3922..3941)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4735..5323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5224..5243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5380..5407
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5408..5435
                     /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 gcgtggtcaa accaagagaa tgaaaactgt aaaacacccc
      181 ccattgactc aactctttaa aatatcacga ttttccattc agaccttata tgcaaaaaag
      241 tttttttttt gtcctgtctt atacaccgag attaatggtg atccttgaaa gctaattatg
      301 gtactgaaag gcttcagata catttctgct aaaaataaac tggaggcaga atagtgccct
      361 ggattctcaa ttgcttacat ctggttccta tttctgagaa acacgtttgg ttttcagaca
      421 gttcattcac agccattgga ggtcctgatt cctgtccccc aacggggaga gcttgtgaca
      481 tcaagcaggt tttgatggat ccgttgtgaa cctatcctct aatagctcca gccagatctg
      541 tggatctcta ttttcatgtg gatgctgcaa ttttctgcac gagtagatga gaaacacttc
      601 atcagctcta atctaagcat tctgggacca gaccgtgtgg tgatggtgcg ctacaaccag
      661 gttcagagga gaaactggcg agttataaca tagaagccac gagctcgggc tgggcataaa
      721 agtcagggca gagccatcta ttgcttacat ttgcttctgg gatccagatc tttcgaagct
      781 agcgctaccg gtcgccacca tggtctccaa gggcgaggag ctgttcaccg gggtggtgcc
      841 catcctggtc gagctggacg gcgatgtcaa cggccacaag ttcagcgtgt ccggcgaggg
      901 cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct
      961 gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcgtgcagt gcttcgcccg
     1021 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1081 ccaggagcgc accatcttct tcaaagacga cggcaactac aagacccgcg ccgaggtgaa
     1141 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1201 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     1261 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     1321 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1381 gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga
     1441 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1501 ggacgagctg tacaagggct ctggtgctac caacttctca ctgttgaaac aggcagggga
     1561 tgtagaggag aatccagggc ctggtgctag tggagactac aaagaccatg acggagatta
     1621 taaagatcat gacatcgatt acaaggatga cgatgacaag tccggactca gatctggagg
     1681 ctccggaggc ccagagccag cgaagtctgc tcccgccccg aaaaagggct ccaagaaggc
     1741 ggtgactaag gcgcagaaga aaggcggcaa gaagcgcaag cgcagccgca aggagagcta
     1801 ttccatctat gtgtataagg ttctgaagca ggtccaccct gacaccggca tttcgtccaa
     1861 ggccatgggc atcatgaact cgtttgtgaa cgacattttc gagcgcatcg caggtgaggc
     1921 ttcccgcctg gcgcattaca acaagcgctc gaccatcacc tccagggaga tccagacggc
     1981 cgtgcgcctg ctgctgcctg gggagttggc caagcacgcc gtgtccgagg gtactaaggc
     2041 catcaccaag tacaccagcg ctaagtaatg aggcgcgccg cggccgcgaa ttcgatatca
     2101 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2161 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2221 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     2281 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2341 cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2401 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2461 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2521 aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc
     2581 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2641 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2701 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctat cagcacacaa ttgcccatta
     2761 tacgcgcgta taatggacta ttgtgtgctg atatgatgcg gtattttctc cttacgcatc
     2821 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     2881 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     2941 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3001 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3061 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3121 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3181 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3241 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3301 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3361 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3421 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3481 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3541 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3601 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3661 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3721 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3781 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3841 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3901 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3961 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4021 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4081 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4141 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4201 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4261 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4321 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4381 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4441 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4501 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4561 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4621 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4681 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4741 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4801 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4861 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4921 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4981 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5041 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5101 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5161 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5221 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5281 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5341 ctttttacgg ttcctggcct tttgctggcc ttttgctcat atcagcacac aattgcccat
     5401 tatacgcgcg tataatggac tattgtgtgc tgataacatg t
//
