LOCUS       sequence-476786-        4968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2430m 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 4968)
  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 4968)
  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: AiP1747 - pAAV-AiE2430m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4968
                     /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      980..989
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(986..988,989..991,992..1702)
                     /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(1031..1052)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1292..1311)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1639..1660
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1795..1815)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2002..2113
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2231..2371
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2231..2360
                     /label=AAV2 ITR
     rep_origin      2446..2901
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2533..2552)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2743..2764
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2918..2937)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3037..3059
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3097..3115)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3183..3287
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3288..3356,3357..4148)
                     /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(3506..3525)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4319..4907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4808..4827
                     /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 gcgttgtaag agtaataatc gcacagaaat tctgtttgaa
      181 cctgggaaat gggtgtaaga acagagggta cccaatggca tctctgaata ggtgacaaca
      241 ctctccgggc atttaaatct gtgagtacgg tgctcctcct ccagtgccgg gaatattgga
      301 cacaaagagc cagcccaggc ctgtgggtcc tgcactctta gagcttttca ttaggcagac
      361 tggctccctg ccatatgcca tctgagtgtc tgcttccatt ccctgggagg agccctgcag
      421 tctgaagcag aactcaggaa tcaaattagc caggcctctg ccgttccctg gtccccaccg
      481 gccaaggcga gttcatcaag gcctcacgct aattcttatc ttgtttaaaa ggggcaaaag
      541 tgactccgca gtttcatctg aggcaattta caccccgcca ctggccgcag ttccaagttc
      601 cttctgtctg ttgtgccatc aaatccatta ttaggacatt tcttcggggg aaacttgatt
      661 gctgttctat cagagaggct cccgggcaat ctatctccat ctgaggccac ggcctgctcg
      721 gctccctcta atcctgttat ccgagggact aattctctaa tgcacacgct gcatacaatg
      781 tatccgattt cctcctaacc caattggatg ccccaaagga ttccagtact gaacacagaa
      841 atgaaggctt agggcccgtg ggacagggtc tccaggcttt ctcccggagc tcgggctggg
      901 cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc cagatctttc
      961 gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     1021 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1081 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     1141 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt
     1201 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1261 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1321 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1381 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1441 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     1501 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1561 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     1621 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1681 cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca
     1741 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1801 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1861 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     1921 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     1981 cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2041 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2101 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2161 aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca cgtgcggacc
     2221 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2281 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2341 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     2401 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     2461 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2521 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2581 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2641 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2701 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2761 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     2821 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2881 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     2941 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3001 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3061 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3121 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3181 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3241 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3301 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3361 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3421 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3481 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3541 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3601 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3661 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3721 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3781 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3841 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3901 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3961 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4021 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4081 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4141 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4201 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4261 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4321 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4381 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4441 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4501 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4561 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4621 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4681 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4741 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4801 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4861 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4921 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
