LOCUS       sequence-461854-        4775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0646h is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in specific populations of brain cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4775)
  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 4775)
  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: AiP12685 - pAAV-AiE0646h-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4775
                     /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      787..796
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(793..795,796..798,799..1509)
                     /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(838..859)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1099..1118)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1446..1467
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1602..1622)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1809..1920
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2038..2178
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2038..2167
                     /label=AAV2 ITR
     rep_origin      2253..2708
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2340..2359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2550..2571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2725..2744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2844..2866
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2904..2922)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2990..3094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3095..3163,3164..3955)
                     /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(3313..3332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4126..4714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4615..4634
                     /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 gcgtacaccc gggctcacta ttagttcccc tacatccacc
      181 ccgcagcctg ctcttaacct ctcctccccg gcggctcaga cccaattctc agtgtcctta
      241 gcgccccctt cgggccacag cgttaagcca cgccccccgg gcccctcatc cgttgccaag
      301 ttcgctgagc gtccgcgtct ggttgcctct ccgccccaca gggcatgttc gtgctgggcc
      361 taccctacgc catcctgcac ggcggctacc tggggttgtt tctcatcatc ttcgccgccg
      421 ttgtgtgctg ctacaccggc aagatcctca tcgcgtgcct gtacgaggag aatgaagacg
      481 gcgaggtggt gcgcgtgcgg gactcgtacg tggccatagc caacgcctgc tgcgccccgc
      541 gcttcccaac gctgggcggc cgagtggtga acgtagcgca gatcatcgag ctggtgatga
      601 cgtgcatcct gtacgtggtg gtgagtggca acctcatgta caacagcttc ccggggctgc
      661 ccgtgtcgca gaagtcctgg tccattatcg ccagagctcg ggctgggcat aaaagtcagg
      721 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
      781 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      841 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      901 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
      961 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     1021 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1081 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1141 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1201 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1261 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1321 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1381 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     1441 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1501 ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     1561 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1621 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1681 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     1741 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagag
     1801 atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1861 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     1921 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     1981 ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag cggccgcagg
     2041 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2101 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     2161 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2221 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2281 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     2341 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2401 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2461 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2521 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2581 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     2641 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     2701 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     2761 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2821 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2881 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2941 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3001 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3061 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3121 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3181 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3241 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3301 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3361 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3421 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3481 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3541 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3601 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3661 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3721 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3781 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3841 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3901 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3961 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4021 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4081 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4141 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4201 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4261 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4321 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4381 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4441 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4501 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4561 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4621 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4681 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4741 gttcctggcc ttttgctggc cttttgctca catgt
//
