LOCUS       sequence-446785-        5685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1426m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal Sst_Chodl neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5685)
  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 5685)
  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: AiP15050 -
            pAAV-AiE1426m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5685
                     /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      1038..1047
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1044..1046,1047..1049,1050..1760)
                     /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(1350..1369)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1697..1718
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1770..1826
                     /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             1839..1904
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1935..2312
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2399..2419)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2606..2717
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2835..2975
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2835..2964
                     /label=AAV2 ITR
     misc_feature    2982..3009
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    3010..3037
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3106..3561
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3193..3212)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3403..3424
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3578..3597)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3697..3719
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3757..3775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3843..3947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3948..4016,4017..4808)
                     /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(4166..4185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4979..5567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5468..5487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5624..5651
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5652..5679
                     /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 gcgtattatg ctaatatgcc ataattaaca taaattaaca
      181 tagttctttg tgggacccca gttaattcat gtgagaaaac ataaaatgtc agggtcgcgg
      241 cgaggggccg gtaggggtgg gcggaggttt ctgaggaggg agcttttggg agcttcgagt
      301 caacaataaa ggaccgaggg ttcggagcag gagtggcctc ggtaaatgaa ttcagatata
      361 attcaagctt attagagggc tttttaaaaa aataaaaaat tgattcagtg tatgaaagca
      421 agttaactac ttctgttcac cattgagaga cttacaggtg cttgcctgaa ctgcaataaa
      481 ggactcttat tattgatcag gacttattta tctcttcatt ttggagagcc tcataaactg
      541 ttattacagt ttctctactg actttgaaaa gtttgaagtt tgaaagagac acctgtgcag
      601 ctaacaaagc atgagcacgg ccagaacaga gaaccctgtt atcatgggtc tatccagtca
      661 gaacggccag ctgaggggcc ctgtgaaagc cagtgctggc cctggaggag ggggcacaca
      721 gccacagatg aaccagttga agaacaccaa cacaatcaat aacggcactc cgcagcaagc
      781 ccagagcatg gccgccacga ttaagcatgt ctgatgttca taaaggccag aagagtttgt
      841 tgaagccact gaacttaggt tataaatgat gacctggtga tgactggaaa gacattaaaa
      901 ctccctccaa aggatctaag aatcaaaact ctggatgtga cctcgagctc gggctgggca
      961 taaaagtcag ggcagagcca tctattgctt acatttgctt ctgggatcca gatctttcga
     1021 agctagcgct accggtcgcc accatggtct ccaagggcga ggagctgttc accggggtgg
     1081 tgcccatcct ggtcgagctg gacggcgatg tcaacggcca caagttcagc gtgtccggcg
     1141 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca
     1201 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcgtg cagtgcttcg
     1261 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1321 acgtccagga gcgcaccatc ttcttcaaag acgacggcaa ctacaagacc cgcgccgagg
     1381 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1441 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1501 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1561 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1621 ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca
     1681 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1741 gcatggacga gctgtacaag ggctctggtg ctaccaactt ctcactgttg aaacaggcag
     1801 gggatgtaga ggagaatcca gggcctggtg ctagtggaga ctacaaagac catgacggag
     1861 attataaaga tcatgacatc gattacaagg atgacgatga caagtccgga ctcagatctg
     1921 gaggctccgg aggcccagag ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga
     1981 aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga
     2041 gctattccat ctatgtgtat aaggttctga agcaggtcca ccctgacacc ggcatttcgt
     2101 ccaaggccat gggcatcatg aactcgtttg tgaacgacat tttcgagcgc atcgcaggtg
     2161 aggcttcccg cctggcgcat tacaacaagc gctcgaccat cacctccagg gagatccaga
     2221 cggccgtgcg cctgctgctg cctggggagt tggccaagca cgccgtgtcc gagggtacta
     2281 aggccatcac caagtacacc agcgctaagt aatgaggcgc gccgcggccg cgaattcgat
     2341 atcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2401 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2461 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg
     2521 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     2581 attccgtggc tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2641 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2701 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2761 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg
     2821 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2881 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2941 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctatcagcac acaattgccc
     3001 attatacgcg cgtataatgg actattgtgt gctgatatga tgcggtattt tctccttacg
     3061 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3121 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3181 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3241 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3301 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     3361 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3421 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     3481 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3541 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     3601 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3661 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3721 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3781 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5101 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5581 cggccttttt acggttcctg gccttttgct ggccttttgc tcatatcagc acacaattgc
     5641 ccattatacg cgcgtataat ggactattgt gtgctgataa catgt
//
