LOCUS       sequence-446820-        5380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1191m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in striatal MSNs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5380)
  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 5380)
  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: AiP14428 -
            pAAV-AiE1191m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5380
                     /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      733..742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(739..741,742..744,745..1455)
                     /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(1045..1064)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1392..1413
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1465..1521
                     /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             1534..1599
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1630..2007
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2094..2114)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2301..2412
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2530..2670
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2530..2659
                     /label=AAV2 ITR
     misc_feature    2677..2704
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2705..2732
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2801..3256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2888..2907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3098..3119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3273..3292)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3392..3414
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3452..3470)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3538..3642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3643..3711,3712..4503)
                     /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(3861..3880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4674..5262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5163..5182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5319..5346
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5347..5374
                     /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 gcgtcacaca aagcctgtct cggttttgtt ttgttttgtt
      181 ttgtcctccc tgtgtcgcat ccacaagtcc caagcgtgta gctcctggga agctatgtgg
      241 ctgttttcac caaggatgac ttctcagaga ctgacaccca taagagaggc ccctccaccc
      301 gagggtggga ggactctctg tggcatcacc gggaggggag gccaggtgtg ggcactgcag
      361 ctatttctaa tccaccagag gaaagagagc tttgactgct tgaaaagcag gctgattgtc
      421 aggggcggaa cccagctcag agctgtcagg gaagctcctg gcagggggtg gagacagggg
      481 gcttggactg acaggcctgt gattttgaaa gaaatgtctt taagagagtg acttaaagta
      541 ggagagaaat attattatta ggtaatgcct gtcaggttat atattttttt ttgatgtgtt
      601 cttggtattt cataatcatt ttgcctttag atggtaagcg agctcgggct gggcataaaa
      661 gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct ttcgaagcta
      721 gcgctaccgg tcgccaccat ggtctccaag ggcgaggagc tgttcaccgg ggtggtgccc
      781 atcctggtcg agctggacgg cgatgtcaac ggccacaagt tcagcgtgtc cggcgagggc
      841 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
      901 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc
      961 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1021 caggagcgca ccatcttctt caaagacgac ggcaactaca agacccgcgc cgaggtgaag
     1081 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1141 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
     1201 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
     1261 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1321 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
     1381 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1441 gacgagctgt acaagggctc tggtgctacc aacttctcac tgttgaaaca ggcaggggat
     1501 gtagaggaga atccagggcc tggtgctagt ggagactaca aagaccatga cggagattat
     1561 aaagatcatg acatcgatta caaggatgac gatgacaagt ccggactcag atctggaggc
     1621 tccggaggcc cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg
     1681 gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat
     1741 tccatctatg tgtataaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag
     1801 gccatgggca tcatgaactc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct
     1861 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc
     1921 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc
     1981 atcaccaagt acaccagcgc taagtaatga ggcgcgccgc ggccgcgaat tcgatatcat
     2041 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2101 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2161 atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc
     2221 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2281 gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2341 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2401 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2461 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac gtgcggaccg
     2521 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     2581 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2641 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctatc agcacacaat tgcccattat
     2701 acgcgcgtat aatggactat tgtgtgctga tatgatgcgg tattttctcc ttacgcatct
     2761 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     2821 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     2881 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2941 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3001 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3061 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3121 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     3181 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3241 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3301 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3361 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3421 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3481 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3541 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3601 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3661 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3721 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3781 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3841 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3901 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3961 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4021 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4081 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4141 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4201 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4261 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4321 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4381 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4441 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4501 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4561 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4621 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4681 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4741 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4801 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4861 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4921 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4981 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5041 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5101 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5161 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5221 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5281 tttttacggt tcctggcctt ttgctggcct tttgctcata tcagcacaca attgcccatt
     5341 atacgcgcgt ataatggact attgtgtgct gataacatgt
//
