LOCUS       sequence-461938-        5008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2271m 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 5008)
  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 5008)
  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: AiP1604 - pAAV-AiE2271m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5008
                     /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      1020..1029
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1026..1028,1029..1031,1032..1742)
                     /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(1071..1092)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1332..1351)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1679..1700
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1835..1855)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2042..2153
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2271..2411
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2271..2400
                     /label=AAV2 ITR
     rep_origin      2486..2941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2573..2592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2783..2804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2958..2977)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3077..3099
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3137..3155)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3223..3327
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3328..3396,3397..4188)
                     /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(3546..3565)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4359..4947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4848..4867
                     /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 gccggcgaac tgctatttct
      181 cagaggaaga taagtctggc tgtaaggaaa tgtgacttaa acataaggct ggtctgattc
      241 ctgccccaca tccctcacca aatctctttg gagggaaaga gcccttaagt ctgtttttcc
      301 agtttgttga gtttgaagtg ggctcagaga tgtgtgctta ggctatacaa atcagcatta
      361 ttctggcctt tgttgtagac acagcaagga ctgatggatg gctatttctg gaagagagga
      421 gggggttact caggaggcgt taactcaaaa gaacaggcca gcctttcttg tggggagggg
      481 agctacactg ttccacctca atgctttatt gagaacccct gatactaagc aaccggggca
      541 cggccttaag aagccaatgg gcaaatcctc tggcagaggt tgaagggaaa gaggcaaggt
      601 ccatgtaggc gggggtgggg gtaggggttt agatggagcc atttaaaagg aaacagaaag
      661 acagcaagag tgacaagcaa tggcaggatc tggtttcagc tggccttgcc aagtaaaccc
      721 tgggaagccc tgtggagaaa ggactctgtt taagttctct aaagtgtttg tggtgctctg
      781 gccaatcaca ttcctaggct gtttctatgg ggcttaccga aggacagaat agtgtcatga
      841 tggccccctc ccttcccatt ctgtaccctt ttctcagcct catgaaataa gtctctggct
      901 tcagtagagc tgtgtaaacg ccggcggagc tcgggctggg cataaaagtc agggcagagc
      961 catctattgc ttacatttgc ttctgggatc cagatctttc gaagctagcg ctaccggtcg
     1021 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1081 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1141 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     1201 ccaccctcgt gaccaccctg ggctacggcg tgcagtgctt cgcccgctac cccgaccaca
     1261 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1321 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1381 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1441 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga
     1501 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     1561 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1621 accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     1681 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1741 agtaagtcga cggcgcgccg cggccgcgaa ttcgatatca taatcaacct ctggattaca
     1801 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     1861 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     1921 ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc
     1981 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggctcga gagatcttcg
     2041 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2101 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2161 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2221 tgggaagaca atagcaggca tgagatctca cgtgcggacc gagcggccgc aggaacccct
     2281 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     2341 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2401 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2461 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2521 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     2581 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2641 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2701 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2761 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     2821 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     2881 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     2941 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3001 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3061 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3121 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3181 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3241 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3301 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3361 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3421 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3481 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3541 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3601 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3661 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3721 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3781 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3841 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3901 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3961 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4021 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4081 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4141 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4201 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4261 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4321 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4381 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4441 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4501 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4561 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4621 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4681 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4741 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4801 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4861 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4921 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4981 gccttttgct ggccttttgc tcacatgt
//
