LOCUS       sequence-450093-        5755 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 5755)
  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 5755)
  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: AiP12546 -
            pAAV-AiE0050m_3xC2-minBG-tdTomato-W...
FEATURES             Location/Qualifiers
     source          1..5755
                     /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      1056..1065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1062..2492
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGIGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(2582..2602)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2789..2900
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3018..3158
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3018..3147
                     /label=AAV2 ITR
     rep_origin      3233..3688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3320..3339)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3530..3551
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3705..3724)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3824..3846
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3884..3902)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3970..4074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4075..4143,4144..4935)
                     /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(4293..4312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5106..5694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5595..5614
                     /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 gcggcgccac catggtgagc aagggcgagg
     1081 aggtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg
     1141 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     1201 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca
     1261 tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt
     1321 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga
     1381 ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg
     1441 gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     1501 ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga
     1561 agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc
     1621 ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg
     1681 aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgg
     1741 ggcatggcat cggcagcacc ggcagcggca gctccggcac cgcctcctcc gaggacaaca
     1801 acatggccgt catcaaagag ttcatgcgct tcaaggtgcg catggagggc tccatgaacg
     1861 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1921 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc
     1981 agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc gattacaaga
     2041 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc
     2101 tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac aaggtgaaga
     2161 tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg
     2221 aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag atccaccagg
     2281 ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc tacatggcca
     2341 agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac atcacctccc
     2401 acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc caccacctgt
     2461 tcctgtacgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc
     2521 gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2581 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2641 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg
     2701 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2761 acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt
     2821 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2881 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2941 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt
     3001 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3061 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     3121 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     3181 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     3241 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3301 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3361 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3421 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3481 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3541 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     3601 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3661 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3721 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3781 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3841 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3901 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3961 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4021 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4081 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4141 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4201 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4261 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4321 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4381 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4441 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4501 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4561 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4621 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4681 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4741 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4801 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4861 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4921 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4981 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5041 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5101 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5161 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5221 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5281 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5341 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5401 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5461 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5521 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5581 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5641 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5701 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
