LOCUS       sequence-450549-        5562 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 5562)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types (preprint).
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 5562)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP12293 -
            pAAV-3xSP10ins-AiE0219h_3xC2-minRho...
FEATURES             Location/Qualifiers
     source          1..5562
                     /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      1149..1158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1155..1157,1158..1160,1161..1871)
                     /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(1200..1221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1461..1480)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1808..1829
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2389..2409)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2596..2707
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2825..2965
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2825..2954
                     /label=AAV2 ITR
     rep_origin      3040..3495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3127..3146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3337..3358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3512..3531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3631..3653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3691..3709)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3777..3881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3882..3950,3951..4742)
                     /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(4100..4119)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4913..5501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5402..5421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgccggtac cgaagctacc cctaacacac tattctacac
      181 acagaaaatg ctcttcacta ggaagctacc cctaacacac tattctacac acagaaaatg
      241 ctcttcacta ggaagctacc cctaacacac tattctacac acagaaaatg ctcttcacta
      301 gacgcgttgg tcttgatcaa ttgtcccagc agagttggtg catgggaaag cctttcttgg
      361 ccttatcagc agctcagcct gggaggttgg cccgggggct gaccaagcac aaaatagaaa
      421 agccactaat aagatccagc tggaacaaat agcctctatt tattttagga aactaatgat
      481 atgtcatcat gacacatatc taaccttagt atttctgatt ggacaggcca gttatggtct
      541 tgatcaattg tcccagcaga gttggtgcat gggaaagcct ttcttggcct tatcagcagc
      601 tcagcctggg aggttggccc gggggctgac caagcacaaa atagaaaagc cactaataag
      661 atccagctgg aacaaatagc ctctatttat tttaggaaac taatgatatg tcatcatgac
      721 acatatctaa ccttagtatt tctgattgga caggccagtt atggtcttga tcaattgtcc
      781 cagcagagtt ggtgcatggg aaagcctttc ttggccttat cagcagctca gcctgggagg
      841 ttggcccggg ggctgaccaa gcacaaaata gaaaagccac taataagatc cagctggaac
      901 aaatagcctc tatttatttt aggaaactaa tgatatgtca tcatgacaca tatctaacct
      961 tagtatttct gattggacag gccagttaga gctcgattca gccgggagct tagggagggg
     1021 aggtcacttc ataagggctt ggggggggag ttggagccac gagtcgtcca gccggagccc
     1081 cgtgtggctg tgctccggcc tcagaagcat ccccggatcc agatctttcg aagctagcgc
     1141 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1201 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1261 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     1321 tgccctggcc caccctcgtg accaccctgg gctacggcgt gcagtgcttc gcccgctacc
     1381 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1441 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1501 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1561 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     1621 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     1681 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1741 tgcccgacaa ccactacctg agctaccagt ccaagctgag caaagacccc aacgagaagc
     1801 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1861 agctgtacaa gtaaactagt gcggccttaa agagaccggt tcactgtgac agtaaaagag
     1921 accggttcac tgtgagaatg aaagagaccg gttcactgtg atcggaaaag agaccggttc
     1981 actgtgagcg gccttaaaga gaccggttca ctgtgacagt aaaagagacc ggttcactgt
     2041 gagaatgaaa gagaccggtt cactgtgatc ggaaaagaga ccggttcact gtgagcggcc
     2101 ttgaaaccca gcagacaatg tagctcagta gaaacccagc agacaatgta gctgaatgga
     2161 aacccagcag acaatgtagc ttcggagaaa cccagcagac aatgtagctg cggccttgaa
     2221 acccagcaga caatgtagct cagtagaaac ccagcagaca atgtagctga atggaaaccc
     2281 agcagacaat gtagcttcgg agaaacccag cagacaatgt agctgcggcc atcataatca
     2341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2461 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     2521 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggc
     2581 tcgagagatc ttcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2641 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2701 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2761 gcaaggggga ggattgggaa gacaatagca ggcatgagat ctcacgtgcg gaccgagcgg
     2821 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2881 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2941 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     3001 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     3061 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     3121 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3181 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3241 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3301 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3361 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     3421 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3481 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     3541 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3601 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3661 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3721 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3781 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3841 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3901 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3961 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4021 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4081 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4141 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4201 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4261 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4321 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4381 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4441 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4501 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4561 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4621 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4681 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4741 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4801 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4861 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4921 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4981 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5041 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5101 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5161 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5221 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5281 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5341 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5401 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5461 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5521 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
