LOCUS       sequence-470627-        5080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE0968m 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 5080)
  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 5080)
  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: AiP13305 -
            pAAV-AiE0968m-minBG-SYFP2-P2A-3XFLA...
FEATURES             Location/Qualifiers
     source          1..5080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     170..188
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(226..248)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     348..367
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(384..839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(521..542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     733..752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   914..1054
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   925..1054
                     /label=AAV2 ITR
     polyA_signal    1172..1283
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1470..1490
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1577..1954)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     CDS             complement(1985..2050)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(2063..2119)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(2129..2839,2840..2842,2843..2845))
                     /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(2171..2192)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2520..2539
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      2842..2851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3267..3396
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3458..4046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3538..3557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4217..5008,5009..5077))
                     /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     4840..4859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5078..5080,1..102))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
       61 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
      121 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
      181 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
      241 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
      301 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
      361 gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt
      421 aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta
      481 taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc
      541 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
      601 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact
      661 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
      721 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc
      781 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta
      841 ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      901 cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca
      961 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     1021 gagggagtgg ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtgagat
     1081 ctcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
     1141 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
     1201 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
     1261 tggctggcaa ctagaaggca cagtcgaaga tctctcgagc cacggaattg tcagtgccca
     1321 acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa
     1381 gaactaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
     1441 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
     1501 taccagtcaa tctttcacaa attttgtaat ccagaggttg attatgatat cgaattcgcg
     1561 gccgcggcgc gcctcattac ttagcgctgg tgtacttggt gatggcctta gtaccctcgg
     1621 acacggcgtg cttggccaac tccccaggca gcagcaggcg cacggccgtc tggatctccc
     1681 gggaggtgat ggtcgagcgc ttgttgtaat gcgccaggcg ggaagcctct cctgcgatgc
     1741 gctcgaaaat gtcgttcaca aacgaattca tgatgcccat ggccttggac gaaatgccgg
     1801 tgtcagggtg gacctgcttc agaaccttgt acacatagat ggaatagctc tccttgcggc
     1861 tgcgcttgcg cttcttgccg cctttcttct gcgccttagt caccgccttc ttggagccct
     1921 ttttcggggc gggagcagac ttcgctggct ctgggcctcc ggagcctcca gatctgagtc
     1981 cggacttgtc atcgtcatcc ttgtaatcga tgtcatgatc tttataatct ccgtcatggt
     2041 ctttgtagtc tccactagca ccaggccctg gattctcctc tacatcccct gcctgtttca
     2101 acagtgagaa gttggtagca ccagagccct tgtacagctc gtccatgccg agagtgatcc
     2161 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     2221 tcagcttgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2281 tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc
     2341 ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt
     2401 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     2461 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     2521 tcttgtagtt gccgtcgtct ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     2581 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca
     2641 cgccgtagcc cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     2701 agatcagctt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     2761 tgaacttgtg gccgttgaca tcgccgtcca gctcgaccag gatgggcacc accccggtga
     2821 acagctcctc gcccttggag accatggtgg cgaccggtag cgctagcttc gaaagatctg
     2881 gatcccagaa gcaaatgtaa gcaatagatg gctctgccct gacttttatg cccagcccga
     2941 gctctatcct aaaagattct actacttgtt taaaaagatg aagtctgagt accactaatt
     3001 ttcagaattt attcttagga tctaggtact gttctgcctc taaatttaat cttcacctca
     3061 atgcttctct acatcacagt taaataactc ctagaacagt tccgctctcc cttgcgatct
     3121 gtgtgcttag ggtgctgcca agctgaattc agggagcagc tgtgttttaa actctgagag
     3181 gttatacctt tagatagatg agatgggatc tcttggagat atgggagtct gcacactgca
     3241 ggagatgtgt ctacgcgtgc ggccgcagga acccctagtg atggagttgg ccactccctc
     3301 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg
     3361 cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa
     3421 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3481 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3541 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3601 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3661 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3721 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3781 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3841 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3901 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3961 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4021 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4081 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4141 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4201 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4261 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4321 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4381 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4441 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4501 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4561 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4621 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4681 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4741 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4801 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4861 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4921 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4981 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5041 agggaataag ggcgacacgg aaatgttgaa tactcatact
//
