LOCUS       sequence-466193-        4592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2185m 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 4592)
  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 4592)
  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: AiP1502 - pAAV-AiE2185m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      157..745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     646..665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   807..936
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      1410..1419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1416..1418,1419..1421,1422..2132)
                     /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(1461..1482)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1722..1741)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2069..2090
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2225..2245)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2432..2543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2661..2801
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2661..2790
                     /label=AAV2 ITR
     rep_origin      2876..3331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2963..2982)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3173..3194
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3348..3367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3467..3489
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3527..3545)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3613..3717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3718..3786,3787..4578)
                     /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(3936..3955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
       61 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
      121 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
      181 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
      241 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
      301 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
      361 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
      421 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
      481 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
      541 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
      601 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
      661 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
      721 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
      781 cttttgctgg ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga
      841 ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
      901 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt tgtaagagta
      961 ataatcgccg gcgaagctag ccccacgcgg gtgtggggtg acaggtttcc agaccacagt
     1021 tacagtgcta ctgtggctga tgtccactag taaaggactg gatgcccgct attggctgta
     1081 aagcaaagac ctggcagtaa agtccccata gggaaggagt acaagctggt gggtagatag
     1141 gcagaatctt agaaaacatt ggctcttggg gacgcggttt agacagctct tgagttggcg
     1201 tggtaacatc acggaagaca aacacgtgag agagggaacg gtgcagaaag cttcaagtgg
     1261 gaggccctgc cccttcacac tagagtttga aggcagagtt gactagtccg ccggcggagc
     1321 tcgggctggg cataaaagtc agggcagagc catctattgc ttacatttgc ttctgggatc
     1381 cagatctttc gaagctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
     1441 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1501 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     1561 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcg
     1621 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1681 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1741 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1801 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1861 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     1921 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1981 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga
     2041 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2101 ggatcactct cggcatggac gagctgtaca agtaagtcga cggcgcgccg cggccgcgaa
     2161 ttcgatatca taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2221 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2281 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     2341 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2401 ctgacaattc cgtggctcga gagatcttcg actgtgcctt ctagttgcca gccatctgtt
     2461 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2521 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2581 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgagatctca
     2641 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     2701 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2761 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     2821 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2881 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2941 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3001 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3061 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3121 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3181 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3241 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3301 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3361 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3421 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3481 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3541 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3601 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3661 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3721 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3781 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3841 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3901 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3961 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4021 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4081 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4141 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4201 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4261 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4321 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4381 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4441 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4501 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4561 ctgattaagc attggtaact gtcagaccaa gt
//
