LOCUS       sequence-470439-        4728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0398m 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 4728)
  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 4728)
  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: AiP12091 - pAAV-AiE0398m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   98..227
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(289..877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(369..388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1048..1839,1840..1908))
                     /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     1671..1690
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1909..2013)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2081..2099
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2137..2159)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2259..2278
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2295..2750)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2432..2453)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2644..2663
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2825..2965
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2836..2965
                     /label=AAV2 ITR
     polyA_signal    3083..3194
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3381..3401
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(3494..4204,4205..4207,4208..4210))
                     /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(3536..3557)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3885..3904
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4144..4165
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4207..4216
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gagaccccag gaagggtcat gtggccaccc tgggcctcag tttcccgata gtgtagtgga
       61 ggagcagtca ctgagggctg tgaacgcgtg cggccgcagg aacccctagt gatggagttg
      121 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
      181 cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa
      241 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
      301 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
      361 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
      421 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
      481 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      541 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      601 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      661 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      721 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      781 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      841 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      901 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      961 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     1021 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     1081 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     1141 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     1201 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     1261 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     1321 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     1381 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     1441 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     1501 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     1561 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     1621 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1681 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     1741 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     1801 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1861 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     1921 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     1981 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     2041 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     2101 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2161 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2221 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     2281 gagagtgcac cataaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     2341 ttaaatcagc tcatttttta accaatagac cgaaatcggc aaaatccctt ataaatcaaa
     2401 agaatagccc gagatagagt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2461 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2521 tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2581 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2641 ggaagggaag aaagcgaaag gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct
     2701 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg
     2761 ctttgacgta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     2821 cgcccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
     2881 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     2941 gccaactcca tcactagggg ttcctgcggc cgctcggtcc gcacgtgaga tctcatgcct
     3001 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
     3061 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
     3121 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
     3181 actagaaggc acagtcgaag atctctcgag ccacggaatt gtcagtgccc aacagccgag
     3241 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc
     3301 aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag
     3361 gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca
     3421 atctttcaca aattttgtaa tccagaggtt gattatgata tcgaattcgc ggccgcggcg
     3481 cgccgtcgac ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     3541 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggt
     3601 agctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     3661 ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg
     3721 ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac gttgtggctg ttgtagttgt
     3781 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     3841 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     3901 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     3961 agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca ctgcacgccg tagcccaggg
     4021 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatc agcttcaggg
     4081 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     4141 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     4201 tgctcaccat ggtggcgacc ggtagcgcta gcttcgaaag atctggatcc cagaagcaaa
     4261 tgtaagcaat agatggctct gccctgactt ttatgcccag cccgagctca accggtcctg
     4321 ggagagagtg tgctggcttg agtgactctg gtttctcgtg tgtgtctgtt tcagatagag
     4381 aaagggacag cttggtggct ttgggcttcc tggagatcta gacatggtgt acctcccccc
     4441 gccccatgtt ctaaaatatc cagtgcagga attccaaggt gcctcaccag ccctcctttg
     4501 gcacagagag gggtggtgtg tggtggtggt atggccagcc caggcgcctg gagcctgtgt
     4561 ctcttgcccc tttgtctcct gtgacacaat gggtgacttc ttatccccat aaagtctctt
     4621 tcagtgtagg gagaagggcc aggatggggg tggaggttga gctaggccag gctatgctag
     4681 gccaagttct gcatgccctc atctgcctag ctgtgccccg agtgaggg
//
