LOCUS       sequence-462004-        4938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2509m 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 4938)
  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 4938)
  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: AiP1857 - pAAV-AiE2509m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1600..1609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1606..1608,1609..1611,1612..2322)
                     /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(1651..1672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1912..1931)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2259..2280
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2415..2435)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2622..2733
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2851..2991
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2851..2980
                     /label=AAV2 ITR
     rep_origin      3066..3521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3153..3172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3363..3384
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3538..3557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3657..3679
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3717..3735)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3803..3907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3908..3976,3977..4768)
                     /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(4126..4145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgttgtaag agtaataatc gttttgtccc tttttcccca ggaacctgac
      841 aggcctgtgc tttctttaat taccactgcc taccaaaatt tccttaccgc tgcttaccaa
      901 aatttccctg tacgagccac cacttggcac agcctgcact caggcggcgc aaagacaaga
      961 atgacacaga caccaagatg ggttcacgca tcttccgcat cttccacagc ttccttttta
     1021 cttgtagctc actccacagc ttttttcact tacagctttt gcagctgttt tttttactta
     1081 gggcaagggc tatatttata cagtagcgtc aaatgacctc atctcgctcc acaccacctc
     1141 atccaatcag aatcacacat gttccaggca cgagctcagg tcggtcacag ataggctgac
     1201 agatccggat catgaggaat aaccatcttg gcccatggga ctgcacccac ttcccgcagg
     1261 gtgccaccat gagcatggat acaagacatc aacaagagaa gcgactgtcc tgctggtaca
     1321 tcttcaggaa gtttatagca gaattagaat tgttacaggg tacacatcat gtaactgtta
     1381 taggctatac aaaatatgac agttacaggt tacacaacat ataactgcta taggctacac
     1441 aatatgtaac tgttataggc tacacaacat gtaactgtta caagctatac atcatataac
     1501 agttcggagc tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc
     1561 ttctgggatc cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc
     1621 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1681 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1741 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1801 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1861 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1921 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1981 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2041 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     2101 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     2161 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     2221 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2281 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg
     2341 cggccgcgaa ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact
     2401 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2461 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta
     2521 gttcttgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2581 ctgttgggca ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca
     2641 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2701 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2761 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2821 tgagatctca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     2881 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     2941 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     3001 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     3061 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3121 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     3181 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3241 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     3301 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3361 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     3421 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     3481 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     3541 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     3601 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3661 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     3721 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     3781 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3841 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3901 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3961 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4021 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4081 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4141 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4201 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4261 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4321 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4381 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4441 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4501 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4561 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4621 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4681 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4741 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4801 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4861 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4921 aaagatcaaa ggatcttc
//
