LOCUS       sequence-470593-        4682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0733m 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 4682)
  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 4682)
  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: AiP13099 - pAAV-AiE0733m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     90..108
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(146..168)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     268..287
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(304..759)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(441..462)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     653..672
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   834..974
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   845..974
                     /label=AAV2 ITR
     polyA_signal    1092..1203
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1390..1410
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1503..2213,2214..2216,2217..2219))
                     /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(1545..1566)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1894..1913
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2153..2174
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2216..2225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2789..2918
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2980..3568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3060..3079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3739..4530,4531..4599))
                     /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     4362..4381
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4600..4682,1..22))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
       61 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
      121 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
      181 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
      241 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
      301 ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
      361 cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg
      421 agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
      481 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
      541 ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
      601 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
      661 aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
      721 ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat
      781 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag
      841 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
      901 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      961 cactaggggt tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg ctattgtctt
     1021 cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac
     1081 tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt
     1141 ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca
     1201 cagtcgaaga tctctcgagc cacggaattg tcagtgccca acagccgagc ccctgtccag
     1261 cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa gaactaacca ggatttatac
     1321 aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg cattaaagca
     1381 gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa tctttcacaa
     1441 attttgtaat ccagaggttg attatgatat cgaattcgcg gccgcggcgc gccgtcgact
     1501 tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     1561 accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactggta gctcaggtag
     1621 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     1681 gcgagctgca cgccgccgtc ctcgatgttg tggcggatct tgaagttggc cttgatgccg
     1741 ttcttctgct tgtcggcggt gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     1801 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     1861 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     1921 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     1981 ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agcccagggt ggtcacgagg
     2041 gtgggccagg gcacgggcag cttgccggtg gtgcagatca gcttcagggt cagcttgccg
     2101 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     2161 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     2221 gtggcgaccg gtagcgctag cttcgaaaga tctggatccc agaagcaaat gtaagcaata
     2281 gatggctctg ccctgacttt tatgcccagc ccgagctctg tgcatgtcta ctagtgagct
     2341 ccctatatag ccactgtaaa ggcaagtggc ctcatcccat gggctggagg cccaggctgc
     2401 aaacacacaa agctttcatt tcactctgct tcctgacagt ggacagacac agtgtaacat
     2461 aatgtagcaa aatgtaacta gttgtccaca tgcgtgcttc caggacttca ccatcatagt
     2521 agtctgtact tgaaactctg agacagaaca agccccatcc ctccccccta cccccacaca
     2581 caacaccagt tgttcttgga aagtattttg tcaagccagt gatgtaacca ggttgacatc
     2641 attggtccca ctgatctttg tgctcatata tcctaagaac tggaggcttt gatagccagc
     2701 cacacttttg gccatccaca ttatggccct aggacttggc cctgactggg cttctgcatg
     2761 tagctctgtg gtgtacgcgt gcggccgcag gaacccctag tgatggagtt ggccactccc
     2821 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2881 ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca
     2941 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3001 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3061 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3121 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3181 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3241 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3301 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3361 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3421 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3481 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3541 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3601 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3661 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3721 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3781 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3841 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3901 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3961 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4021 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4081 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4141 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4201 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4261 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4321 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4381 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4441 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4501 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4561 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4621 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4681 aa
//
