LOCUS       sequence-446777-        5728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1425m 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 5728)
  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 5728)
  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: AiP15049 -
            pAAV-AiE1425m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5728
                     /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      1081..1090
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1087..1089,1090..1092,1093..1803)
                     /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(1393..1412)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1740..1761
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1813..1869
                     /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             1882..1947
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1978..2355
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2442..2462)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2649..2760
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2878..3018
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2878..3007
                     /label=AAV2 ITR
     misc_feature    3025..3052
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    3053..3080
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      3149..3604
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3236..3255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3446..3467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3621..3640)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3740..3762
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3800..3818)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3886..3990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3991..4059,4060..4851)
                     /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(4209..4228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5022..5610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5511..5530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5667..5694
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5695..5722
                     /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 gcgtcctaga accagtttcc aagtcacccc cagcaaatct
      181 gcacctccag attacaagcc tgcccccaga cacttcactt cctaacaacc accagtacct
      241 ccagattata agcctgcccc cagacacttc acttcctaac aaccaccagt caagaggaaa
      301 tcagaagtca agtttgtggt ttggctctca gcaccagtca attatgttaa aggctataat
      361 ggccctgcca tcaaatgttt gtcaggctag ataccccctt cttgcttcta taaatgcttg
      421 actaaaattc ttagcctctc tgtcattgga ctagccagat cttagcctat aatacacact
      481 gataaaatac ctaattaagt acccagaaca cacttttcag tttgctgaca tctgcagtta
      541 gtggaggtct tctgttagca tagaaagcac taaaaattca cgatgtgacc catttcagaa
      601 cttactgaga taaatacatt tgattatctt attcacctcc tgtgaatcaa gacatttagt
      661 aacataaaac tctggcaatt cgtggagaaa tacggaaaat aactgctggc tggttgcttc
      721 tagcattttt agataaatta acaaagaaaa aactgagccc catgataaaa ttaagtaatc
      781 ccatgataca gtgtaaaagg gggaaaatga acttggtgat ataatcaaca ggatccaggt
      841 gcagataaat agtctaaatg tttctaagtg tgccctagaa aataaaattc tttccatcag
      901 ctatggagct caagttcagg aaaatcaaag accttactat aattttggat gaattacaat
      961 gaaaattgaa gtctcagcct caaaagggag ctcgggctgg gcataaaagt cagggcagag
     1021 ccatctattg cttacatttg cttctgggat ccagatcttt cgaagctagc gctaccggtc
     1081 gccaccatgg tctccaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1141 ctggacggcg atgtcaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1201 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
     1261 cccaccctcg tgaccaccct gggctacggc gtgcagtgct tcgcccgcta ccccgaccac
     1321 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1381 atcttcttca aagacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1441 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1501 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     1561 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     1621 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1681 aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     1741 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1801 aagggctctg gtgctaccaa cttctcactg ttgaaacagg caggggatgt agaggagaat
     1861 ccagggcctg gtgctagtgg agactacaaa gaccatgacg gagattataa agatcatgac
     1921 atcgattaca aggatgacga tgacaagtcc ggactcagat ctggaggctc cggaggccca
     1981 gagccagcga agtctgctcc cgccccgaaa aagggctcca agaaggcggt gactaaggcg
     2041 cagaagaaag gcggcaagaa gcgcaagcgc agccgcaagg agagctattc catctatgtg
     2101 tataaggttc tgaagcaggt ccaccctgac accggcattt cgtccaaggc catgggcatc
     2161 atgaactcgt ttgtgaacga cattttcgag cgcatcgcag gtgaggcttc ccgcctggcg
     2221 cattacaaca agcgctcgac catcacctcc agggagatcc agacggccgt gcgcctgctg
     2281 ctgcctgggg agttggccaa gcacgccgtg tccgagggta ctaaggccat caccaagtac
     2341 accagcgcta agtaatgagg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     2401 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     2461 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     2521 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     2581 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagag
     2641 atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2701 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2761 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2821 ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag cggccgcagg
     2881 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2941 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3001 cgcgcagctg cctgcagggg cgcctatcag cacacaattg cccattatac gcgcgtataa
     3061 tggactattg tgtgctgata tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3121 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     3181 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     3241 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3301 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3361 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3421 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3481 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     3541 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3601 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3661 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3721 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3781 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3841 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3901 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3961 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4021 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4081 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4141 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4201 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4261 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4321 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4381 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4441 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4501 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4561 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4621 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4681 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4741 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4801 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4861 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4921 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4981 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5041 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5101 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5161 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5221 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5281 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5341 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5401 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5461 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5521 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5581 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5641 ctggcctttt gctggccttt tgctcatatc agcacacaat tgcccattat acgcgcgtat
     5701 aatggactat tgtgtgctga taacatgt
//
