LOCUS       sequence-454363-        5115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0050m_3xC2 is an enhancer sequence, designed to drive 
            AAV-mediated transgene expression in cortical L4/5_IT neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5115)
  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 5115)
  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: AiP13152 -
            pAAV-AiE0050m_3xC2-minBG-iCre(R297T...
FEATURES             Location/Qualifiers
     source          1..5115
                     /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      1051..1060
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1063..1083
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2149..2260
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2378..2518
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2378..2507
                     /label=AAV2 ITR
     rep_origin      2593..3048
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2680..2699)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2890..2911
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3065..3084)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3184..3206
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3244..3262)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3330..3434
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3435..3503,3504..4295)
                     /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(3653..3672)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4466..5054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4955..4974
                     /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 gcgtaggcaa cttgttaaaa atctcagtca aacttacata
      181 ctatatagaa cagcatggtg aatttagggc acatggatat aaaatggaag tttcttattc
      241 agtagcagca acttgtgggc acaggagttg gcaaagataa aaatgtccaa agtcacaaat
      301 acaatgtata gttagtcata ggtgctgtta tttgcctcaa aaaatagact tttattttgc
      361 ctttcttttc tttaaccaca ctcaaaatta gagaacagag acaaaaccca gcaggaaata
      421 ggcaacttgt taaaaatctc agtcaaactt acatactata tagaacagca tggtgaattt
      481 agggcacatg gatataaaat ggaagtttct tattcagtag cagcaacttg tgggcacagg
      541 agttggcaaa gataaaaatg tccaaagtca caaatacaat gtatagttag tcataggtgc
      601 tgttatttgc ctcaaaaaat agacttttat tttgcctttc ttttctttaa ccacactcaa
      661 aattagagaa cagagacaaa acccagcagg aaataggcaa cttgttaaaa atctcagtca
      721 aacttacata ctatatagaa cagcatggtg aatttagggc acatggatat aaaatggaag
      781 tttcttattc agtagcagca acttgtgggc acaggagttg gcaaagataa aaatgtccaa
      841 agtcacaaat acaatgtata gttagtcata ggtgctgtta tttgcctcaa aaaatagact
      901 tttattttgc ctttcttttc tttaaccaca ctcaaaatta gagaacagag acaaaaccca
      961 gcaggaaatg agctcgggct gggcataaaa gtcagggcag agccatctat tgcttacatt
     1021 tgcttctggg atccagatct ttcgaagcta gccaccatgg tgcccaagaa gaagaggaaa
     1081 gtctccaacc tgctgactgt gcaccaaaac ctgcctgccc tccctgtgga tgccacctct
     1141 gatgaagtca ggaagaacct gatggacatg ttcagggaca ggcaggcctt ctctgaacac
     1201 acctggaaga tgctcctgtc tgtgtgcaga tcctgggctg cctggtgcaa gctgaacaac
     1261 aggaaatggt tccctgctga acctgaggat gtgagggact acctcctgta cctgcaagcc
     1321 agaggcctgg ctgtgaagac catccaacag cacctgggcc agctcaacat gctgcacagg
     1381 agatctggcc tgcctcgccc ttctgactcc aatgctgtgt ccctggtgat gaggagaatc
     1441 agaaaggaga atgtggatgc tggggagaga gccaagcagg ccctggcctt tgaacgcact
     1501 gactttgacc aagtcagatc cctgatggag aactctgaca gatgccagga catcaggaac
     1561 ctggccttcc tgggcattgc ctacaacacc ctgctgcgca ttgccgaaat tgccagaatc
     1621 agagtgaagg acatctcccg caccgatggt gggagaatgc tgatccacat tggcaggacc
     1681 aagaccctgg tgtccacagc tggtgtggag aaggccctgt ccctgggggt taccaagctg
     1741 gtggagagat ggatctctgt gtctggtgtg gctgatgacc ccaacaacta cctgttctgc
     1801 cgggtcagaa agaatggtgt ggctgcccct tctgccacct cccaactgtc cacccgggcc
     1861 ctggaaggga tctttgaggc cacccaccgc ctgatctatg gtgccaagga tgactctggg
     1921 cagagatacc tggcctggtc tggccactct gccagagtgg gtgctgccac cgacatggcc
     1981 agggctggtg tgtccatccc tgaaatcatg caggctggtg gctggaccaa tgtgaacatt
     2041 gtgatgaact acatcagaaa cctggactct gagactgggg ccatggtgag gctgctcgaa
     2101 gatggggact gaggcgcgcc gaattcaagc ttctcgagag atcttcgact gtgccttcta
     2161 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2221 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2281 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2341 gcaggcatga gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     2401 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2461 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     2521 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     2581 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2641 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     2701 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2761 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2821 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2881 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     2941 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     3001 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3061 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3121 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3181 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3241 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3301 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3361 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3421 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3481 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3541 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3601 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3661 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3721 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3781 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3841 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3901 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3961 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4021 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4081 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4141 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4201 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4261 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4321 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4381 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4441 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4501 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4561 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4621 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4681 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4741 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4801 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4861 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4921 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4981 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5041 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5101 cttttgctca catgt
//
