LOCUS       sequence-474064-        5046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0277h 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 5046)
  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 5046)
  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: AiP11791 -
            pAAV-hsA2-AiE0277h-minRho-SYFP2-WPR...
FEATURES             Location/Qualifiers
     source          1..5046
                     /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      1083..1092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1089..1091,1092..1094,1095..1805)
                     /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(1134..1155)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1395..1414)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1742..1763
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1873..1893)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2080..2191
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2309..2449
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2309..2438
                     /label=AAV2 ITR
     rep_origin      2524..2979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2611..2630)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2821..2842
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2996..3015)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3115..3137
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3175..3193)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3261..3365
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3366..3434,3435..4226)
                     /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(3584..3603)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4397..4985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4886..4905
                     /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 gcggccgcaa cgcgtttaga acaatggctg gcccatagta aatgccgtgt
      181 tagtgtgtta gttgctgttc ttccacgtca gaagaggcac agacaaatta ccaccaggtg
      241 gcgctcagag tctgcggagg catcacaaca gccctgaatt tgaatcctgc tctgccactg
      301 cctagttgag accttttact acctgactag ctgtttgtgt attttaggtg tttgtttgtg
      361 tattttaggt gtttgtttag agagtcatgg agattagcga ttttgttcag tgatagctga
      421 agatcacagg atgctcagtt aaattctaat aatttcagta aaattataat agtttgtcta
      481 ctaaaaatca ttttgcttga tagctctggg aacttagaaa tttcatggga aaattatttt
      541 tctccctttc agagaataga aagcaaagcc agtaaacagt caggccttct gccaaaggaa
      601 aattatctat gcctagaatg gataaattcc cagctcataa agattttttt cttcatctcc
      661 acggcatagt cacatcttat tctaagagcc tctggcagtt gctggaaaat tgttggacca
      721 ctgggtatgt cattaagacc atcttcaccc ttaaaaagaa ttgtacatca aagtagtgca
      781 tgcacagtga aacttctcat gatatctaaa gcctgaaaat aattattcag aatataggag
      841 agaataaatt tatacctgtt gagttgctaa tgcatgctag ggagtgtgct gaattcgata
      901 tcataatcaa ccataggtac cgagctcggg attcagccgg gagcttaggg aggggaggtc
      961 acttcataag ggcctggggg gggagttgga gccacgagtc gtccagccgg agccccgtgt
     1021 ggctgagctc cggcctcaga agcatccccg ggttggatcc ttcgaagcta gcgctaccgg
     1081 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1141 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1201 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     1261 ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc
     1321 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1381 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1441 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1501 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     1561 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     1621 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1681 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1741 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1801 acaagtaagt cgacatcata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1861 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1921 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt
     1981 tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2041 gttgggcact gacaattccg tggctcgaga gatcttcgac tgtgccttct agttgccagc
     2101 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2161 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2221 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2281 agatctcacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     2341 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2401 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     2461 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     2521 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2581 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2641 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2701 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2761 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2821 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     2881 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     2941 ttaacgggaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     3001 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3061 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3121 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3181 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3241 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3301 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3361 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3421 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3481 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3541 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3601 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3661 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3721 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3781 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3841 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3901 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3961 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4021 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4081 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4141 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4201 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4261 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4321 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4381 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4441 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4501 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4561 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4621 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4681 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4741 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4801 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4861 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4921 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4981 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5041 acatgt
//
