LOCUS       sequence-432576-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2443m 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 5126)
  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 5126)
  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: AiP1869 - pAAV-AiE2443m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..5126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     198..219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(373..392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     492..514
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(552..570)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        638..742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(743..811,812..1603)
                     /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(961..980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1774..2362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2263..2282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2424..2553
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      3561..3570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3567..3569,3570..3572,3573..4283)
                     /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(3612..3633)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3873..3892)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4220..4241
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4376..4396)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4583..4694
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4812..4952
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4812..4941
                     /label=AAV2 ITR
     rep_origin      join(5027..5126,1..356)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(join(5114..5126,1..7))
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
       61 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
      121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
      181 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
      241 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
      301 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
      361 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
      421 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
      481 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
      541 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
      601 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      661 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      961 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1261 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1441 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1681 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1741 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1801 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1861 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1921 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1981 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2041 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2101 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2161 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2221 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2281 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2341 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2401 ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc
     2461 cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     2521 agggagtggc caactccatc actaggggtt cctgcggccg cacgcgttgt aagagtaata
     2581 atcgactggc caatggtctc acctgttcta cctccccatg gctgggtttc cagttgtgca
     2641 cgggaactca agacttttac ataagtccta gggatgcaac ctagtgtcct catgcttcca
     2701 tggcattccc tttaagaact aagctatctc cacagccctg ctttgagttg tcaatctttc
     2761 attctctgca aagaatctat tagatgagtt cagctttgtc ctggttagaa caaaaggaag
     2821 tatgccttgt ttgtgatcct gcagaatgga gagggttggg aaactccata tactttactg
     2881 acaaataaag caaacatgaa acactgaaat cccatatgtt gtgaggcacg ctggcagtct
     2941 gcctcatccc caacccagtg caagcaggct ggtttgcaat ctggagcctc cctcgctgag
     3001 ctgtaacaaa attgatccta ggtgcatgta agacacagcg cattccttcc cgtgctgatt
     3061 tatccctggc ctcttcccta catttctcct tttagcccat catttagtct ctgatataac
     3121 tcacaggcaa ccattgaaaa taatgcaggg ggttacttct cagcagcagc agtggtggag
     3181 atccttctgc ctgtatgagc aaagaggcca tgtggagcca gcaagccttc ccaacgaagg
     3241 tcaggtgtaa agcgtagatt accaacagca ctaggaagac aagcactgca ttgatgtcat
     3301 tcaccctaat agccccaccc cctggcacca tgcctggaat gagagggata ctaaaaaaat
     3361 catttatgga atgactcaag aaaaggcaaa ggagttggct catccagtaa agtgcttacc
     3421 tagcaaacct gagaacctga gctcagtccc aagaacccat atcaacggag ctcgggctgg
     3481 gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat ccagatcttt
     3541 cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     3601 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3661 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
     3721 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct
     3781 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3841 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3901 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3961 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     4021 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     4081 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4141 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc
     4201 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     4261 tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga attcgatatc
     4321 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4381 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4441 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     4501 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4561 ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4621 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4681 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4741 caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac
     4801 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4861 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4921 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     4981 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     5041 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5101 gccttagcgc ccgctccttt cgcttt
//
