LOCUS       sequence-461991-        4425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2052m_C2 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 4425)
  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 4425)
  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: AiP1418 - pAAV-AiE2052m_C2-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4425
                     /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      437..446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(443..445,446..448,449..1159)
                     /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(488..509)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(749..768)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1096..1117
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1252..1272)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1459..1570
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1688..1828
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1688..1817
                     /label=AAV2 ITR
     rep_origin      1903..2358
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1990..2009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2200..2221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2375..2394)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2494..2516
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2554..2572)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2640..2744
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2745..2813,2814..3605)
                     /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(2963..2982)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3776..4364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4265..4284
                     /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 gcgttaagga ccagctccta ctgaggacat aaatgttatg
      181 tttcttaaaa gtctatacca taagaatctt cttcagatct tggtgtgaag attcttaccc
      241 cagattggtt acagaagcgg taccaggtgg gtggatccca tcaaaagctt tagtgagaaa
      301 tcaccctttt ccttcacagc agcccagctg actggtgaac tttgagctcg ggctgggcat
      361 aaaagtcagg gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa
      421 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      481 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      541 gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa
      601 gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc
      661 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      721 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      781 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      841 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
      901 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
      961 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1021 cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa
     1081 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1141 catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa
     1201 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     1261 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     1321 ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat
     1381 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     1441 ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1501 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     1561 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     1621 acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag
     1681 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     1741 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     1801 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     1861 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     1921 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     1981 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2041 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2101 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2161 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2221 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     2281 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     2341 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2401 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2461 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2521 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     2581 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2641 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2701 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2761 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2821 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2881 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2941 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3001 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3061 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3121 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3181 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3241 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3301 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3361 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3421 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3481 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3541 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3601 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3661 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3721 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3781 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3841 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3901 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3961 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4021 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4081 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4141 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4201 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4261 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4321 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4381 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
