LOCUS       sequence-438968-        4945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AiE2133m 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 4945)
  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 4945)
  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: AiP1426 - pAAV-AiE2133m-minBG-SYFP2-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..4945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     470..489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(708..812)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     880..898
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(936..958)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1058..1077
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1094..1549)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1231..1252)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1443..1462
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1624..1764
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1635..1764
                     /label=AAV2 ITR
     polyA_signal    1882..1993
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2180..2200
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2293..3003,3004..3006,3007..3009))
                     /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(2335..2356)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2684..2703
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2943..2964
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3006..3015
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3842..3971
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4033..4621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4113..4132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4792..4945,1..638,639..707))
                     /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"
ORIGIN
        1 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
       61 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
      121 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
      181 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
      241 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
      301 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
      361 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
      421 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
      481 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
      541 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
      601 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
      661 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
      721 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
      781 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
      841 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
      901 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      961 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     1021 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     1081 agagtgcacc ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     1141 taaatcagct cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa
     1201 gaatagcccg agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     1261 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     1321 gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     1381 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     1441 gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg
     1501 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc
     1561 tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     1621 gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc
     1681 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     1741 ccaactccat cactaggggt tcctgcggcc gctcggtccg cacgtgagat ctcatgcctg
     1801 ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc cagaatagaa
     1861 tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag gacagtggga
     1921 gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga tggctggcaa
     1981 ctagaaggca cagtcgaaga tctctcgagc cacggaattg tcagtgccca acagccgagc
     2041 ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa gaactaacca
     2101 ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat gatacaaagg
     2161 cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa taccagtcaa
     2221 tctttcacaa attttgtaat ccagaggttg attatgatat cgaattcgcg gccgcggcgc
     2281 gccgtcgact tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     2341 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactggta
     2401 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     2461 gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg tggcggatct tgaagttggc
     2521 cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctgt tgtagttgta
     2581 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     2641 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     2701 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     2761 gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgccgt agcccagggt
     2821 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatca gcttcagggt
     2881 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     2941 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     3001 gctcaccatg gtggcgaccg gtagcgctag cttcgaaaga tctggatccc agaagcaaat
     3061 gtaagcaata gatggctctg ccctgacttt tatgcccagc ccgagctccg ccggcgctgc
     3121 ggggcagtgg ggtgcactgg ttctgatctc cggcgacctc tgatctttgt aaactcacaa
     3181 atgtgctcag gagcctgagg aggccagaaa ggagacgtct caggggttgg aggctgggca
     3241 gatggcaggt caggtgggat ggagctgggc agagactggc ctccctgctg gagaaggtga
     3301 cgtggctgtg cttcctgtgg ggcctgtgga ggcttcctcc agggcttctt aggtaagacg
     3361 gcttcctgtg cactcgtctc aggccatgaa gtcagccctc cacatatctg gcctgaggac
     3421 agagaaggct tcctctttgg ggagtgaaaa ggaacgtggc ctcccaagct tcaggaagga
     3481 aagccctcac ttcctggtga gagggaacaa acacggcagg ccttgactcc aaacccagag
     3541 gaacacgtca gagcctgccg cctgggacag agacagttgg ctggcctagt cctgtggctg
     3601 tgcttgccga cgtcctgacc ttacgatgga ggcaagcctg tgaacgccat gggggtagga
     3661 aaagcagagg attttcctgg gggtcacttg gaagcctagg tctagatgtc taagccgctg
     3721 tgccctgcag agcctggctc ctgcaggtgc tcagagatcc atgttcaaag actgtgagcc
     3781 acaggcacgc tctatgctct tccgcgccgg cgattattac tcttacaacg cgtgcggccg
     3841 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3901 gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3961 ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4021 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4081 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4141 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4201 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4441 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4561 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4621 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4681 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4741 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4801 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4861 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4921 aatgataccg cgagacccac gctca
//
