LOCUS       sequence-432570-        4745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2531m 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 4745)
  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 4745)
  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: AiP1852 - pAAV-AiE2531m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(160..179)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     279..301
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(339..357)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        425..529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(530..598,599..1390)
                     /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(748..767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1561..2149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2050..2069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2211..2340
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2967..2976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2973..2975,2976..2978,2979..3689)
                     /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(3018..3039)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3279..3298)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3626..3647
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3782..3802)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3989..4100
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4218..4358
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4218..4347
                     /label=AAV2 ITR
     rep_origin      join(4433..4745,1..143)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4520..4539)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(4730..4745,1..6)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
       61 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
      121 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
      181 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      241 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
      301 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
      361 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
      421 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      481 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      541 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      601 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      661 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      721 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      781 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      841 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      901 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
      961 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1021 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1081 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1141 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1201 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1261 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1321 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1381 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1441 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1501 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1561 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1621 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1681 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1741 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1801 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1861 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1921 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1981 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2041 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2101 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2161 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
     2221 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
     2281 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     2341 gcggccgcac gcgttgtaag agtaataatc gatgagagta taataaaaca caatctcaga
     2401 cactgatcca gtggcagcta ttttaaggca gcgattatag caaaatgaga cactgattta
     2461 taacacattg ttccttctgg ccccagggca aattgcatgg tttgccccag aggagactta
     2521 agtgcgtcgc tgagaaaaga acttgcttcg attgtggctt tagaacaatc cagtgggcat
     2581 tgtctccatg tgcgtgctaa cagagaatgc aggcttctat tgcaaagtaa ttatctctta
     2641 aagttattga tgtctttctg gtagctgccc aagtgcattg aaagaatcac acagctatta
     2701 gcacagttca atataaaagt cctcccactg aagacagctt tattgatatc gcacatccca
     2761 aactgaaatc agaattccag tcagaagcca ccagaagctt ggcttggcag gagcactttg
     2821 tgactcagta tctaacgcta tcttgttgta tttaaccctg tagcacatag ccggagctcg
     2881 ggctgggcat aaaagtcagg gcagagccat ctattgctta catttgcttc tgggatccag
     2941 atctttcgaa gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca
     3001 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     3061 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     3121 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcgtgc
     3181 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     3241 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     3301 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     3361 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     3421 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     3481 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     3541 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
     3601 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3661 tcactctcgg catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc
     3721 gatatcataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3781 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3841 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg
     3901 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3961 acaattccgt ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt
     4021 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4081 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4141 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt
     4201 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4261 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4321 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4381 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4441 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4501 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4561 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4621 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4681 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4741 ttcca
//
