LOCUS       sequence-449779-        4470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0675m is an enhancer sequence, designed to drive AAV-mediated 
            transgene expression in cortical L2/3_IT neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4470)
  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 4470)
  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: AiP12858 - pAAV-AiE0675m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4470
                     /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      482..491
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(488..490,491..493,494..1204)
                     /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(533..554)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(794..813)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1141..1162
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1297..1317)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1504..1615
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1733..1873
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1733..1862
                     /label=AAV2 ITR
     rep_origin      1948..2403
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2035..2054)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2245..2266
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2420..2439)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2539..2561
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2599..2617)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2685..2789
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2790..2858,2859..3650)
                     /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(3008..3027)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3821..4409
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4310..4329
                     /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 cgcgtaagca gagccccttt tgtcagcctt
      181 ctctcttact gttgtggttg tcatcacaac cccctccctc cagtaaaatg aagacacgga
      241 ttctccttgg ttactggggg aggtatccag gatacgccag gcctccagca ccagaataag
      301 acagccaagg cctccagcct tgtacactga agcagctcct ttcctctcta gaacaaagct
      361 agccattcct gaattaccca gaacgcgtga gctcgggctg ggcataaaag tcagggcaga
      421 gccatctatt gcttacattt gcttctggga tccagatctt tcgaagctag cgctaccggt
      481 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      541 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      601 cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg
      661 gcccaccctc gtgaccaccc tgggctacgg cgtgcagtgc ttcgcccgct accccgacca
      721 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      781 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      841 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      901 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca
      961 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     1021 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1081 caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     1141 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1201 caagtaagtc gacggcgcgc cgcggccgcg aattcgatat cataatcaac ctctggatta
     1261 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     1321 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     1381 ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc
     1441 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggctc gagagatctt
     1501 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1561 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1621 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1681 attgggaaga caatagcagg catgagatct cacgtgcgga ccgagcggcc gcaggaaccc
     1741 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     1801 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     1861 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     1921 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     1981 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2041 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2101 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2161 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2221 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2281 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2341 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     2401 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     2461 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     2521 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     2581 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     2641 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2701 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2761 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2821 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2881 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2941 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3001 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3061 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3121 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3181 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3241 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3301 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3361 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3421 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3481 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3541 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3601 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3661 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3721 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3781 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3841 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3901 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3961 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4021 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4081 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4141 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4201 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4261 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4321 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4381 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4441 tggccttttg ctggcctttt gctcacatgt
//
