LOCUS       sequence-449791-        4918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0219h_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical Vip+ neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4918)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle TL, Dwivedi D, Way SW, 
            Oster A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya
            X, Roth JR, Alexander JR, Allen S, Amster A, Arbuckle J, Ayala A, 
            Baker PM, Bakken TE, Barcelli T, Barta S, Bendrick J, Bertagnolli D,
            Bielstein C, Bishwakarma P, Bowlus J, Boyer G, Brouner K, Casian B, 
            Casper T, Chakka AB, Chakrabarty R, Chance RK, Chavan S, Clark M, 
            Colbert K, Collman F, Daniel S, Departee M, DiValentin P, Donadio N,
            Dotson N, Egdorf T, Fliss T, Gabitto M, Garcia J, Gary A, Gasperini 
            M, Gloe J, Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, 
            Halterman C, Haradon Z, Hastings SD, Helback O, Ho W, Hockemeyer D, 
            Huang C, Huff S, Hunker A, Johansen N, Jones D, Juneau Z, Kalmbach 
            B, Kannan M, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, Lee AY, 
            Leibly M, Lenz GH, Li S, Liang E, Lusk N, Madigan Z, Malloy J, 
            Malone J, McCue R, Melchor J, Mollenkopf T, Moosman S, Morin E, 
            Newman D, Ng L, Ngo K, Omstead V, Otto S, Oyama A, Pena N, Pham T, 
            Phillips E, Pom CA, Potekhina L, Ransford S, Ray PL, Rette D, 
            Reynoldson C, Rimorin C, Rocha D, Ruiz A, Sanchez REA, Sawyer L, 
            Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shepard N, Shulga L, 
            Sigler AR, Siverts L, Soliman S, Somasundaram S, Staats B, Stewart 
            K, Szelenyi E, Tieu M, Trader C, Tran A, van Velthoven CTJ, Walker 
            M, Wang Y, Weed N, Wirthlin M, Wood T, Wynalda B, Yao Z, Zhou T, 
            Ariza J, Dee N, Reding M, Ronellenfitch K, Mufti S, Sunkin SM, Smith
            KA, Esposito L, Waters J, Thyagarajan B, Yao S, Lein ES, Zeng H, 
            Levi BP, Ngai J, Ting JT, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   Cell. 2025 May 16:S0092-8674(25)00513-6. doi: 
            10.1016/j.cell.2025.05.002.
  PUBMED    40403729
REFERENCE   2  (bases 1 to 4918)
  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"; journalName: "Cell. 2025 
            May 16:S0092-8674(25)00513-6. doi: 10.1016/j.cell.2025.05.002."
COMMENT     Sequence Label: AiP15265 -
            pAAV-AiE0219h_3xC2-minBG-SYFP2-WPRE...
FEATURES             Location/Qualifiers
     source          1..4918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      930..939
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(936..938,939..941,942..1652)
                     /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(981..1002)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1242..1261)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1589..1610
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1745..1765)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1952..2063
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2181..2321
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2181..2310
                     /label=AAV2 ITR
     rep_origin      2396..2851
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2483..2502)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2693..2714
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2868..2887)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2987..3009
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3047..3065)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3133..3237
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3238..3306,3307..4098)
                     /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(3456..3475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4269..4857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4758..4777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttggtc ttgatcaatt gtcccagcag
      181 agttggtgca tgggaaagcc tttcttggcc ttatcagcag ctcagcctgg gaggttggcc
      241 cgggggctga ccaagcacaa aatagaaaag ccactaataa gatccagctg gaacaaatag
      301 cctctattta ttttaggaaa ctaatgatat gtcatcatga cacatatcta accttagtat
      361 ttctgattgg acaggccagt tatggtcttg atcaattgtc ccagcagagt tggtgcatgg
      421 gaaagccttt cttggcctta tcagcagctc agcctgggag gttggcccgg gggctgacca
      481 agcacaaaat agaaaagcca ctaataagat ccagctggaa caaatagcct ctatttattt
      541 taggaaacta atgatatgtc atcatgacac atatctaacc ttagtatttc tgattggaca
      601 ggccagttat ggtcttgatc aattgtccca gcagagttgg tgcatgggaa agcctttctt
      661 ggccttatca gcagctcagc ctgggaggtt ggcccggggg ctgaccaagc acaaaataga
      721 aaagccacta ataagatcca gctggaacaa atagcctcta tttattttag gaaactaatg
      781 atatgtcatc atgacacata tctaacctta gtatttctga ttggacaggc cagttagagc
      841 tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc
      901 cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
      961 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1021 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     1081 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
     1141 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1201 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1261 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1321 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1381 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     1441 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1501 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
     1561 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1621 ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa
     1681 ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1741 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1801 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     1861 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     1921 ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     1981 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2041 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2101 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     2161 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2221 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2281 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     2341 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2401 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2461 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2521 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2581 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     2641 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2701 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     2761 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2821 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     2881 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2941 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3001 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3061 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3121 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3181 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3241 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3301 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3361 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3421 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3481 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3541 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3601 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3661 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3721 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3781 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3841 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3901 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3961 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4021 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4081 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4141 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4201 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4261 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4321 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4381 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4441 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4501 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4561 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4621 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4681 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4741 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4801 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4861 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
