LOCUS       sequence-466311-        4777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2470m 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 4777)
  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 4777)
  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: AiP1843 - pAAV-AiE2470m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     207..226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(445..549)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     617..635
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(673..695)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     795..814
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(831..1286)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(968..989)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1180..1199
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1361..1501
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1372..1501
                     /label=AAV2 ITR
     polyA_signal    1619..1730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1917..1937
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2030..2740,2741..2743,2744..2746))
                     /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(2072..2093)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2421..2440
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2680..2701
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2743..2752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3411..3540
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3602..4190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3682..3701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4361..4777,1..375,376..444))
                     /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"
ORIGIN
        1 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
       61 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
      121 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
      181 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
      241 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
      301 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
      361 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      421 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
      481 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      541 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
      601 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
      661 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
      721 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
      781 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa
      841 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
      901 atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagagttgag
      961 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     1021 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt
     1081 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     1141 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     1201 gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     1261 gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata
     1321 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct
     1381 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg
     1441 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     1501 tgcggccgct cggtccgcac gtgagatctc atgcctgcta ttgtcttccc aatcctcccc
     1561 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
     1621 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
     1681 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaagatct
     1741 ctcgagccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     1801 aggcggcgat gagttccgcc gtggcaagaa ctaaccagga tttatacaag gaggagaaaa
     1861 tgaaagccat acgggaagca atagcatgat acaaaggcat taaagcagcg tatccacata
     1921 gcgtaaaagg agcaacatag ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca
     1981 gaggttgatt atgatatcga attcgcggcc gcggcgcgcc gtcgacttac ttgtacagct
     2041 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     2101 gcttctcgtt ggggtctttg ctcagcttgg actggtagct caggtagtgg ttgtcgggca
     2161 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     2221 cgccgtcctc gatgttgtgg cggatcttga agttggcctt gatgccgttc ttctgcttgt
     2281 cggcggtgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     2341 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     2401 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     2461 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     2521 ggtagcgggc gaagcactgc acgccgtagc ccagggtggt cacgagggtg ggccagggca
     2581 cgggcagctt gccggtggtg cagatcagct tcagggtcag cttgccgtag gtggcatcgc
     2641 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     2701 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggta
     2761 gcgctagctt cgaaagatct ggatcccaga agcaaatgta agcaatagat ggctctgccc
     2821 tgacttttat gcccagcccg agctccgctc atgtaaaaca taatctcttc tctttctatg
     2881 cagagtccca cctggacttc ctgattctct aattgtcctc ttacaatcta cattcatact
     2941 tactctccat atacaccaat caagttgtgt tactccactg tcagaatgtc ctgcagtggt
     3001 ctcctctcct acctagaaaa atagctggtt ctgttacaac cagtgatgac gggattgtaa
     3061 gctggctgct cccctggttt catcccccac tctgcaacca tcctttgctc tcaccagtct
     3121 ctgtgggctc tgtgctattc ccaccagaca cactcccacc ttgaggcctt tacaccagct
     3181 ggtccctcta cctagaatac actgctccca gattctcaca cagcagactc tgactccagc
     3241 cctcattcaa atatcatctc tagaacaaat ctcggcccct gagcactttg ttgaaactgc
     3301 attctttctc tctcctgcct ttagtaattc cactctaatt tttcctcaat tattacaatt
     3361 attcaattgt atcacatgac gattattact cttacaacgc gtgcggccgc aggaacccct
     3421 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3481 aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3541 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3601 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3661 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3721 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3781 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3841 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3901 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3961 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4021 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4081 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4141 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4201 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4261 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4321 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4381 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4441 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4501 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4561 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4621 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4681 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4741 caaggcgagt tacatgatcc cccatgttgt gcaaaaa
//
