LOCUS       sequence-325642-        4793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0058h is an enhancer sequence designed to drive AAV-mediated 
            transgene expression in L4/5_IT cortical neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4793)
  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 4793)
  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: AiP11402-pAAV-AiE0058h-minBG-SYFP2-WPRE3-BGHpA...
FEATURES             Location/Qualifiers
     source          1..4793
                     /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      805..814
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(811..813,814..816,817..1527)
                     /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(856..877)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1117..1136)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1464..1485
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1620..1640)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1827..1938
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2056..2196
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2056..2185
                     /label=AAV2 ITR
     rep_origin      2271..2726
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2358..2377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2568..2589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2743..2762)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2862..2884
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2922..2940)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3008..3112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3113..3181,3182..3973)
                     /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(3331..3350)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4144..4732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4633..4652
                     /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 gcgtcaaaag atggaagttg ggaggttgaa gaagtgcagg
      181 atggcattcc aagtgatggg ggcaatggca tggaggtagg aaagcataag gtatattcag
      241 gctataaata atagttagat ttggctggat cctggatttg agaagccagg aaatgagata
      301 acactggtca ctttcactaa agctcatgaa aaaaaaaata catacatata tatatatata
      361 aaataaatat acatatatat ttttaagccc catatgacta gaggaggcag cccatctgtt
      421 ctctgggctt cacttttctt gtctgggaaa tgagtaggtt ggactgcatg gtctttaagg
      481 tctctttagt attatcttgt ttgactccgt aaagagaaaa acaaaggttc ctcctgacat
      541 cttgtgttgc cttccaacgt ccagtccagt gtgattgttt taagtactct ttggatattt
      601 tactgttata aaaagtgaag aaaaagactg attttgccaa gtcttatgga tccaaattag
      661 tactcattgc actatggtca tttagttgag gacgatactc cagcttcaaa ggagctcggg
      721 ctgggcataa aagtcagggc agagccatct attgcttaca tttgcttctg ggatccagat
      781 ctttcgaagc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
      841 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      901 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct gatctgcacc
      961 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta cggcgtgcag
     1021 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1081 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1141 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1201 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1261 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1321 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1381 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa gctgagcaaa
     1441 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1501 actctcggca tggacgagct gtacaagtaa gtcgacggcg cgccgcggcc gcgaattcga
     1561 tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1621 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1681 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     1741 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     1801 aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat ctgttgtttg
     1861 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1921 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1981 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga tctcacgtgc
     2041 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2101 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2161 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2221 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2281 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2341 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2401 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2461 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     2521 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2581 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     2641 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2701 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     2761 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2821 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     2881 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     2941 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3001 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3061 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3121 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3181 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3241 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3301 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3361 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3421 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3481 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3541 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3601 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3661 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3721 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3781 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3841 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3901 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3961 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4021 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4081 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4141 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4201 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4261 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4321 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4381 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4441 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4501 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4561 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4621 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4681 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4741 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
