LOCUS       sequence-461900-        5269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0340m 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 5269)
  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 5269)
  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: AiP11992 -
            pAAV-3xSP10ins-AiE0340m-minRho-SYFP...
FEATURES             Location/Qualifiers
     source          1..5269
                     /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      1281..1290
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1287..1289,1290..1292,1293..2003)
                     /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(1332..1353)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1593..1612)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1940..1961
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2096..2116)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2303..2414
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2532..2672
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2532..2661
                     /label=AAV2 ITR
     rep_origin      2747..3202
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2834..2853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3044..3065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3219..3238)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3338..3360
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3398..3416)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3484..3588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3589..3657,3658..4449)
                     /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(3807..3826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4620..5208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5109..5128
                     /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 gcgccggtac cgaagctacc cctaacacac tattctacac
      181 acagaaaatg ctcttcacta ggaagctacc cctaacacac tattctacac acagaaaatg
      241 ctcttcacta ggaagctacc cctaacacac tattctacac acagaaaatg ctcttcacta
      301 gacgcgttac ctgccattcc aactctggcc tctgtgagca tatacacaca catgcgcaca
      361 ttaacatccc ccccccccca cacacacctc aggcatagac atacactttt aaaaaggcag
      421 ccacaatatc aatttaattc agataaaatc acaacacaac cttctttagt actgagtctt
      481 acaaagttgt atgcttgagg tgtcctgaga accccactaa ggtaatggct gtaacattcc
      541 tcagaggtga actcatttcc atgactcatt ctgaaaagca caggctcctt gtcctttcac
      601 acgcatcaca atgcaaccta aaatagatcc tcatcagcgc ctctttcagg gttatgaata
      661 gcactacctt gaaaagggcc aagagggatg catgcatggg attcagaaag gaggcattca
      721 agaaaaccaa gcaggaggca ggctgcctaa gtactttgag catggagcat gacgggaccc
      781 cctctggctc ccacgcaggc aaccaggtgc cacactcaat ggagatggag tgggtgggag
      841 ctctgagaag gagctgcatc tactgggcaa agtggttgaa aaacccgaag aggacctaag
      901 taagcatcgg cttagactgc tgaccccgtt ctccacctct ttatccaccc ccctttctgg
      961 gtggctggaa aaatcaaatg gtatttctct accttcctga ttgtcacatt caatgtaccc
     1021 aacggaagga attcacagtg ggccacctcc tgctcaagaa gccaaggcac tttgcattct
     1081 gcctcttgtc tttgctggga gaatggcatg acaacttaag gagctcgatt cagccgggag
     1141 cttagggagg ggaggtcact tcataagggc ttgggggggg agttggagcc acgagtcgtc
     1201 cagccggagc cccgtgtggc tgtgctccgg cctcagaagc atccccggat ccagatcttt
     1261 cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1321 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1381 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
     1441 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct
     1501 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1561 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1621 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1681 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1741 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     1801 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1861 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc
     1921 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1981 tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga attcgatatc
     2041 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2101 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2161 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     2221 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2281 ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2341 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2401 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2461 caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac
     2521 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2581 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     2641 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     2701 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     2761 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2821 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2881 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2941 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     3001 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3061 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     3121 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3181 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     3241 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3301 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3361 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3421 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3601 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3661 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3721 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3781 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3841 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3901 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3961 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4021 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4081 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4141 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4201 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4261 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4321 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4381 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4441 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4501 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4561 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4621 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4681 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4741 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4801 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4861 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4921 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4981 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5041 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5101 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5161 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5221 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
