LOCUS       sequence-446774-        5390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE1419m 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 5390)
  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 5390)
  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: AiP15043 -
            pAAV-AiE1419m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5390
                     /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      743..752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(749..751,752..754,755..1465)
                     /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(1055..1074)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1402..1423
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1475..1531
                     /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             1544..1609
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1640..2017
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(2104..2124)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2311..2422
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2540..2680
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2540..2669
                     /label=AAV2 ITR
     misc_feature    2687..2714
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    2715..2742
                     /gene="telL"
                     /label=telL
                     /note="telomeric sequence from coliphage N15"
     rep_origin      2811..3266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2898..2917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3108..3129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3283..3302)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3402..3424
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3462..3480)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3548..3652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3653..3721,3722..4513)
                     /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(3871..3890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4684..5272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5173..5192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5329..5356
                     /gene="telR"
                     /label=telR
                     /note="telomeric sequence from coliphage N15"
     misc_feature    5357..5384
                     /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 gcgtaacttg acagtggtgg gcaaaagtaa aacttgaaag
      181 acaaaacttg gaagtgctgg aactgtaact ccagaggtct ctgagagagc ctggcttcga
      241 gaactcttta actccctctt ctcagcctct gagcagcaaa cccagcacag ctgttctcgc
      301 tgtaaagggc agctatgcag ggcagagttc ccagaaggct tctatgctct caaatccgct
      361 ctataaagta ggaagtgtgc tgagtaccag gccatccata ctgacaacca cagcagatcc
      421 accgccaagc atttttattg ttgcggcaga tggtggcctg attgtgaagc gtcccagagc
      481 aagggtgccc ttcaacattg gaaattaata gctaagctac tcatctccct tcaaatcaga
      541 ggcaatgtca ggctccgaga caaccttcag ggtagtttaa ctgagtccgg aagctgccag
      601 agcaggaccc atgtgggcca gccactgtcc ctctttctgt agctgactgg agctcgggct
      661 gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctggg atccagatct
      721 ttcgaagcta gcgctaccgg tcgccaccat ggtctccaag ggcgaggagc tgttcaccgg
      781 ggtggtgccc atcctggtcg agctggacgg cgatgtcaac ggccacaagt tcagcgtgtc
      841 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
      901 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg
      961 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1021 aggctacgtc caggagcgca ccatcttctt caaagacgac ggcaactaca agacccgcgc
     1081 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1141 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1201 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1261 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1321 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga
     1381 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1441 tctcggcatg gacgagctgt acaagggctc tggtgctacc aacttctcac tgttgaaaca
     1501 ggcaggggat gtagaggaga atccagggcc tggtgctagt ggagactaca aagaccatga
     1561 cggagattat aaagatcatg acatcgatta caaggatgac gatgacaagt ccggactcag
     1621 atctggaggc tccggaggcc cagagccagc gaagtctgct cccgccccga aaaagggctc
     1681 caagaaggcg gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa
     1741 ggagagctat tccatctatg tgtataaggt tctgaagcag gtccaccctg acaccggcat
     1801 ttcgtccaag gccatgggca tcatgaactc gtttgtgaac gacattttcg agcgcatcgc
     1861 aggtgaggct tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat
     1921 ccagacggcc gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg
     1981 tactaaggcc atcaccaagt acaccagcgc taagtaatga ggcgcgccgc ggccgcgaat
     2041 tcgatatcat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2101 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2161 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac
     2221 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2281 tgacaattcc gtggctcgag agatcttcga ctgtgccttc tagttgccag ccatctgttg
     2341 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2401 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2461 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gagatctcac
     2521 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     2581 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     2641 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctatc agcacacaat
     2701 tgcccattat acgcgcgtat aatggactat tgtgtgctga tatgatgcgg tattttctcc
     2761 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2821 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2881 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2941 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3001 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3061 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3121 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     3181 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3241 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3301 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3361 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3421 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3481 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3541 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3601 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3661 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3721 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3781 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3841 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3901 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3961 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4021 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4081 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4141 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4201 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4261 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4321 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4381 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4441 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4501 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4561 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4621 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4681 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4741 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4801 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4861 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4921 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4981 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5041 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5101 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5161 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5221 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5281 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcata tcagcacaca
     5341 attgcccatt atacgcgcgt ataatggact attgtgtgct gataacatgt
//
