LOCUS       sequence-470432-        5325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2174m 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 5325)
  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 5325)
  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: AiP1403 - pAAV-AiE2174m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..5325
                     /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      1337..1346
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1343..1345,1346..1348,1349..2059)
                     /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(1388..1409)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1649..1668)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1996..2017
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2152..2172)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2359..2470
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2588..2728
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2588..2717
                     /label=AAV2 ITR
     rep_origin      2803..3258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2890..2909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3100..3121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3275..3294)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3394..3416
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3454..3472)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3540..3644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3645..3713,3714..4505)
                     /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(3863..3882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4676..5264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5165..5184
                     /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 gccggcgtcc agcaaacgat
      181 aatgaatggc tccacaccaa ccatatatgg agctttacaa cagaactatt catgtttcca
      241 gagatattcc tgactcttcc acagatctga agaaaaccca ggcacatccc atttcaccct
      301 ggatgaagtc aacttaatcc tcacccacca atctacactc tttctgaccc agattttgag
      361 gacccagctg atcgtgctgc ctgcgaggac cacattttca tcattgacat aatggttaat
      421 caactgtcct cttcgatcct ccagctgttt gcctcttctc cctcagtctc agcacactgg
      481 gaccagtctt agaacaccag ggctaagcta ttccagaagc acagaccaat tctaataggt
      541 tcctgaaaag tgggtgggca ggggtggagt tagggcacca tgtcgaactc cagctggccg
      601 cccataagca gactgggctc tgaaggacag agcaggctct cagggcaaga agcgccatta
      661 caggaatgga gaggcctatg cgtgctgcat gagggctgca ctgtatcatg tgcagtttga
      721 caggaaacca ctcaggtttg atttcaagat aacaacttga gcttataaca acttcttaga
      781 acctcattga gatgagaaga ccgaagcgga gaaagggatg gtggaggggg aagaacggga
      841 agatgtggta gaggaggaag tagtgaagga agaagaaaag gaggaggggg aggaggggga
      901 ggaggaggag gagcaaggga ggaggaagat gtggtaagga gtagatggaa gaggaagaaa
      961 aggaggagga gggggaggag ggggaggagg aagaggccat ctgcaagtca ctggtagaaa
     1021 gggaagcacc tcatgagcca gaactgtgac tgatcactgg gaagcatgaa cctggtacgg
     1081 ctggatagct gataaccaca gcaaagacca ggcctgggcc ccacttgttc tccgactgct
     1141 ggtccgtgca agcaggggat tgctgaggaa gaggtggggg ctcacagaca tctgtaccac
     1201 aggccacagc ctcagcagat ctgctgatgc tctatcgccg gcggagctcg ggctgggcat
     1261 aaaagtcagg gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa
     1321 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     1381 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1441 gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa
     1501 gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc
     1561 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1621 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1681 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1741 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1801 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     1861 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1921 cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa
     1981 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2041 catggacgag ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa
     2101 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2161 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2221 ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat
     2281 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2341 ggctcgagag atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2401 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2461 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2521 acagcaaggg ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag
     2581 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     2641 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     2701 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     2761 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     2821 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     2881 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2941 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3001 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3061 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3121 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     3181 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3241 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3301 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3361 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3421 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     3481 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3541 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3601 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3661 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3721 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3781 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3841 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3901 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3961 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4021 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4081 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4141 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4201 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4261 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4321 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4381 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4441 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4501 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4561 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4621 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4681 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4741 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4801 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4861 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4921 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4981 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5041 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5101 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5161 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5221 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5281 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
