LOCUS       sequence-470483-        4852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0734m 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 4852)
  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 4852)
  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: AiP12627 - pAAV-AiE0734m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   288..417
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(479..1067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(559..578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1238..2029,2030..2098))
                     /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     1861..1880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2099..2203)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2271..2289
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2327..2349)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2449..2468
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2485..2940)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2622..2643)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2834..2853
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3015..3155
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3026..3155
                     /label=AAV2 ITR
     polyA_signal    3273..3384
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3571..3591
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3684..4394,4395..4397,4398..4400))
                     /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(3726..3747)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4075..4094
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4334..4355
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4397..4406
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ggaaactgct gagtggccta ctctaggggg ccagtgaggg acgaagcctg gattggaccc
       61 tagatgcagc atgtgggtga gagtccaggg ctctggagcc cagctctcca ccctgaggtt
      121 ttccagagat ctatgatgga ggtgggtcct tgggtaggcc ctggtcatgg acagggaacc
      181 ctgggaggaa gggctggcct ctgccccttg tgtggctggg ttctctggat caagtcagtt
      241 accctttctg ggcctcagct gcatctctga tttacgcgtg cggccgcagg aacccctagt
      301 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
      361 ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca
      421 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      481 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      541 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      601 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      661 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      721 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      781 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      841 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      901 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      961 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1021 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1081 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1141 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1201 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1261 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1321 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1381 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1441 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1501 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1561 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1621 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1681 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1741 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1801 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1861 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1921 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1981 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2041 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2101 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2161 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2221 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     2281 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2341 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2401 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     2461 agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt
     2521 taaatttttg ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt
     2581 ataaatcaaa agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc
     2641 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2701 gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2761 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2821 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag
     2881 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2941 actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     3001 ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     3061 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     3121 agagggagtg gccaactcca tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga
     3181 tctcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc
     3241 ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa
     3301 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
     3361 atggctggca actagaaggc acagtcgaag atctctcgag ccacggaatt gtcagtgccc
     3421 aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca
     3481 agaactaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
     3541 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
     3601 ataccagtca atctttcaca aattttgtaa tccagaggtt gattatgata tcgaattcgc
     3661 ggccgcggcg cgccgtcgac ttacttgtac agctcgtcca tgccgagagt gatcccggcg
     3721 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc
     3781 ttggactggt agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
     3841 gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc
     3901 ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac gttgtggctg
     3961 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     4021 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     4081 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     4141 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg
     4201 tagcccaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatc
     4261 agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     4321 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     4381 tcctcgccct tgctcaccat ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc
     4441 cagaagcaaa tgtaagcaat agatggctct gccctgactt ttatgcccag cccgagctct
     4501 gaatgaataa acagatggct gaatggctga atgaatgaat gaatgggtga atggctgaat
     4561 gggtgaatgg ctgaatgggt gagtgctttg agggaaggaa cccaaagcca gtgagctcag
     4621 acttcctcag gatcctgggc agcctgtggt aacttcgctg tttgtctctt gtggtttgcc
     4681 agagcgtctt caaaggctct gaccggagct ggcttctcac caggctcttc ctgtctgtct
     4741 gctcacatgg tgcagttgcc ctcaggcctc taacctccag gggtgcaccc atgtggcaga
     4801 gtgtgtggca gcctagcacc ttgcacgggt acccatcttc ctcttatggt ag
//
