LOCUS       sequence-325688-        4805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2004m 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 4805)
  AUTHORS   Graybuck LT, Daigle TL, Sedeno-Cortes AE, Walker M, Kalmbach B, Lenz
            GH, Morin E, Nguyen TN, Garren E, Bendrick JL, Kim TK, Zhou T, 
            Mortrud M, Yao S, Siverts LA, Larsen R, Gore BB, Szelenyi ER, Trader
            C, Balaram P, van Velthoven CTJ, Chiang M, Mich JK, Dee N, Goldy J, 
            Cetin AH, Smith K, Way SW, Esposito L, Yao Z, Gradinaru V, Sunkin 
            SM, Lein E, Levi BP, Ting JT, Zeng H, Tasic B
  TITLE     Enhancer viruses for combinatorial cell-subclass-specific labeling.
  JOURNAL   Neuron. 2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011.
  PUBMED    33789083
REFERENCE   2  (bases 1 to 4805)
  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: "Neuron. 
            2021 Mar 23. pii: S0896-6273(21)00159-8. doi: 
            10.1016/j.neuron.2021.03.011."
COMMENT     Sequence Label: AiP12014-pAAV-AiE2004m-minCMV-SYFP2-WPRE3-BGHp...
FEATURES             Location/Qualifiers
     source          1..4805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        717..755
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      817..826
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(823..825,826..828,829..1539)
                     /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(868..889)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1129..1148)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1476..1497
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1632..1652)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1839..1950
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2068..2208
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2068..2197
                     /label=AAV2 ITR
     rep_origin      2283..2738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2370..2389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2580..2601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2755..2774)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2874..2896
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2934..2952)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3020..3124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3125..3193,3194..3985)
                     /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(3343..3362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4156..4744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4645..4664
                     /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 gcgtcgcgta taagccttgg gggcaatcaa actattacat
      181 tgagtccttg gatttgctac aaattacatt ttaaatgcaa tcattttata aaagcttcaa
      241 cactcacact tggaagcgtt accctgttga atatcactga ctcactaact tgcattgcca
      301 tgctaacttg ctttcagaga gatctcagaa cacatcatct tctgctattt caatacatgc
      361 acattaattt cctatcaacg tgtgctgatc aggaactctg taatctggca ccggtgttta
      421 tttttattcc tgtctattcc tgttggctca cgaaaagatt gtttgagcaa gtgttttatg
      481 gtgagttgta tcatatgtac attgatttaa tctgcccaca ttcagttcta caagcggagc
      541 caaaaaaata gagacaagca taattttcat tcaacatgag cccctcaatg caagccaagt
      601 acctcatctg gtgctcagct aaagcaacag caatctgttc caccctggag acacaactgg
      661 ccacagaaaa cttagtgaaa agaggcaatg ctatgcacag gacaaatgag ctcagaggta
      721 ggcgtgtacg gtgggaggcc tatataagca gagctggttt agtgaaccgt cagatcgcct
      781 ggggatccag atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag
      841 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      901 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      961 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     1021 tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1081 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1141 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1201 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1261 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1321 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     1381 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1441 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1501 gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg
     1561 ccgcgaattc gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     1621 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1681 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     1741 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     1801 ttgggcactg acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc
     1861 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1921 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1981 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga
     2041 gatctcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     2101 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     2161 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     2221 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2281 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2341 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2401 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2461 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2521 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2581 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     2641 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     2701 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     2761 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     2821 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     2881 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     2941 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3001 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3061 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3121 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3181 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3241 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3301 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3361 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3421 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3481 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3541 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3601 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3661 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3721 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3781 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3841 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3901 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3961 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4021 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4081 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4141 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4201 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4261 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4321 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4381 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4441 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4501 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4561 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4621 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4681 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4741 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4801 catgt
//
