LOCUS       sequence-452845-        5280 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 5280)
  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 5280)
  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..5280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     regulatory      1292..1301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1298..1300,1301..1303,1304..2014)
                     /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(1343..1364)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1604..1623)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1951..1972
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2107..2127)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    2314..2425
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2543..2683
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2543..2672
                     /label=AAV2 ITR
     rep_origin      2758..3213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2845..2864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3055..3076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3230..3249)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3349..3371
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3409..3427)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3495..3599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3600..3668,3669..4460)
                     /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(3818..3837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4631..5219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5120..5139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgccggta ccgaagctac ccctaacaca
      181 ctattctaca cacagaaaat gctcttcact aggaagctac ccctaacaca ctattctaca
      241 cacagaaaat gctcttcact aggaagctac ccctaacaca ctattctaca cacagaaaat
      301 gctcttcact agacgcgtta cctgccattc caactctggc ctctgtgagc atatacacac
      361 acatgcgcac attaacatcc cccccccccc acacacacct caggcataga catacacttt
      421 taaaaaggca gccacaatat caatttaatt cagataaaat cacaacacaa ccttctttag
      481 tactgagtct tacaaagttg tatgcttgag gtgtcctgag aaccccacta aggtaatggc
      541 tgtaacattc ctcagaggtg aactcatttc catgactcat tctgaaaagc acaggctcct
      601 tgtcctttca cacgcatcac aatgcaacct aaaatagatc ctcatcagcg cctctttcag
      661 ggttatgaat agcactacct tgaaaagggc caagagggat gcatgcatgg gattcagaaa
      721 ggaggcattc aagaaaacca agcaggaggc aggctgccta agtactttga gcatggagca
      781 tgacgggacc ccctctggct cccacgcagg caaccaggtg ccacactcaa tggagatgga
      841 gtgggtggga gctctgagaa ggagctgcat ctactgggca aagtggttga aaaacccgaa
      901 gaggacctaa gtaagcatcg gcttagactg ctgaccccgt tctccacctc tttatccacc
      961 cccctttctg ggtggctgga aaaatcaaat ggtatttctc taccttcctg attgtcacat
     1021 tcaatgtacc caacggaagg aattcacagt gggccacctc ctgctcaaga agccaaggca
     1081 ctttgcattc tgcctcttgt ctttgctggg agaatggcat gacaacttaa ggagctcgat
     1141 tcagccggga gcttagggag gggaggtcac ttcataaggg cttggggggg gagttggagc
     1201 cacgagtcgt ccagccggag ccccgtgtgg ctgtgctccg gcctcagaag catccccgga
     1261 tccagatctt tcgaagctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
     1321 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1381 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     1441 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     1501 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1561 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1621 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1681 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1741 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     1801 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     1861 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct
     1921 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1981 cgggatcact ctcggcatgg acgagctgta caagtaagtc gacggcgcgc cgcggccgcg
     2041 aattcgatat cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2101 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2161 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc
     2221 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2281 cactgacaat tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg
     2341 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2401 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2461 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct
     2521 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     2581 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2641 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     2701 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     2761 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2821 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2881 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2941 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3001 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3061 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     3121 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3181 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     3241 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     3301 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3361 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     3421 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3481 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3541 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3601 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3661 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3721 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3781 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3841 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3901 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3961 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4021 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4081 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4141 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4201 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4261 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4321 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4381 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4441 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4501 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4561 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4621 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4681 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4741 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4801 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4861 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4921 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4981 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5041 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5101 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5161 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5221 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
