LOCUS       sequence-471002-        4955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2179m 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 4955)
  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 4955)
  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: AiP1408 - pAAV-AiE2179m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4955
                     /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      967..976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(973..975,976..978,979..1689)
                     /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(1018..1039)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1279..1298)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1626..1647
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1782..1802)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1989..2100
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2218..2358
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2218..2347
                     /label=AAV2 ITR
     rep_origin      2433..2888
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2520..2539)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2730..2751
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2905..2924)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3024..3046
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3084..3102)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3170..3274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3275..3343,3344..4135)
                     /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(3493..3512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4306..4894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4795..4814
                     /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 gcgtgatata tagacctaat ggccacagca aagcaggaaa
      181 cagaagcctt ggaaggcatg tgacctctgc tggcgactct cattgtcagc atgaagaaca
      241 gcaaacccag tacaaccgtt tcctgtgact gtgtcataga tcgattactc aaggctgggt
      301 ggcttgaaac cacaagcata agatctccat tgccagggtc agaaggctgg gaaatagctg
      361 agttctctgg caactcggaa ggagatggaa aggcctgggt tgaggaaatg tgtcacgtac
      421 atgaagtcac aaggcattgt cacgaaaggt agccacacac ggaccactag gtttaagtcg
      481 atgcaaccac ttctgcctta gctgcctgtg tgttctcctc caactcatct cccgctccca
      541 cactgctccc ttgtttcaaa gtcccagaat tcctgagttg tgccacagac agacacctct
      601 gatcgttatt gtccagtctc gtttttcatc ttgaaccgtg cacatacagc atatcatgtt
      661 ggatgtcttc tggaactttc ctttcccctt gacatcctgt ctctgaaaat caacttcatc
      721 cttgcatctg gtttcagttc ctctgtcttt gctgctacac cagtcactgg tgagaaccgg
      781 aaataggtac aatacattgt aacgtatcca cttgaggttt gggtgttatt tttagtatgc
      841 aaatcgggga gggctgtcag taagctctcc tttgagctcg ggctgggcat aaaagtcagg
      901 gcagagccat ctattgctta catttgcttc tgggatccag atctttcgaa gctagcgcta
      961 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1021 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1081 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     1141 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
     1201 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1261 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1321 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1381 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1441 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1501 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1561 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     1621 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1681 ctgtacaagt aagtcgacgg cgcgccgcgg ccgcgaattc gatatcataa tcaacctctg
     1741 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1801 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1861 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     1921 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggctcgagag
     1981 atcttcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2041 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2101 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2161 ggaggattgg gaagacaata gcaggcatga gatctcacgt gcggaccgag cggccgcagg
     2221 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2281 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     2341 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2401 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2461 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     2521 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2581 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2641 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2701 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2761 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     2821 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     2881 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     2941 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3001 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3061 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3121 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3181 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3241 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3301 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3361 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3421 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3481 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3541 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3601 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3661 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3721 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3781 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3841 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3901 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3961 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4021 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4081 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4141 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4201 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4321 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4381 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4441 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4501 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4561 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4621 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4681 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4741 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4801 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4861 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4921 gttcctggcc ttttgctggc cttttgctca catgt
//
