LOCUS       sequence-466357-        4821 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AiE3089m 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 4821)
  AUTHORS   Ben-Simon Y, Hooper M, Narayan S, Daigle T, Dwivedi D, Way SW, Oster
            A, Stafford DA, Mich JK, Taormina MJ, Martinez RA, Opitz-Araya X, 
            Roth JR, Allen S, Ayala A, Bakken TE, Barcelli T, Barta S, Bendrick 
            J, Bertagnolli D, Bowlus J, Boyer G, Brouner K, Casian B, Casper T, 
            Chakka AB, Chakrabarty R, Chance RK, Chavan S, Departee M, Donadio 
            N, Dotson N, Egdorf T, Gabitto M, Garcia J, Gary A, Gasperini M, 
            Goldy J, Gore BB, Graybuck L, Greisman N, Haeseleer F, Halterman C, 
            Helback O, Hockemeyer D, Huang C, Huff S, Hunker A, Johansen N, 
            Juneau Z, Kalmbach B, Khem S, Kussick E, Kutsal R, Larsen R, Lee C, 
            Lee AY, Leibly M, Lenz GH, Liang E, Lusk N, Malone J, Mollenkopf T, 
            Morin E, Newman D, Ng L, Ngo K, Omstead V, Oyama A, Pham T, Pom CA, 
            Potekhina L, Ransford S, Rette D, Rimorin C, Rocha D, Ruiz A, 
            Sanchez REA, Sedeno-Cortes A, Sevigny JP, Shapovalova N, Shulga L, 
            Sigler AR, Siverts LA, Somasundaram S, Stewart K, Szelenyi E, Tieu 
            M, Trader C, van Velthoven CTJ, Walker M, 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 J, Tasic B
  TITLE     A suite of enhancer AAVs and transgenic mouse lines for genetic 
            access to cortical cell types.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244.
  PUBMED    38915722
REFERENCE   2  (bases 1 to 4821)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Sep 26:2024.06.10.597244. doi: 
            10.1101/2024.06.10.597244."
COMMENT     Sequence Label: AiP20046 - pAAV-AiE3089m-minBG-SYFP2-WPRE3-BGHpA
FEATURES             Location/Qualifiers
     source          1..4821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..37
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   199..339
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   210..339
                     /label=AAV2 ITR
     polyA_signal    457..568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     755..775
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(868..1578,1579..1581,1582..1584))
                     /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(910..931)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1259..1278
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1518..1539
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1581..1590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2293..2422
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2484..3072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2564..2583)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3243..4034,4035..4103))
                     /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     3866..3885
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4104..4208)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4276..4294
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4332..4354)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4454..4473
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(4490..4821,1..124))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4627..4648)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt
       61 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
      121 gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa
      181 aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
      241 gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
      301 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgctcg gtccgcacgt
      361 gagatctcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca
      421 ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag
      481 gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca
      541 acagatggct ggcaactaga aggcacagtc gaagatctct cgagccacgg aattgtcagt
      601 gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt
      661 ggcaagaact aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat
      721 agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt
      781 aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattat gatatcgaat
      841 tcgcggccgc ggcgcgccgt cgacttactt gtacagctcg tccatgccga gagtgatccc
      901 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
      961 cagcttggac tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     1021 gggggtgttc tgctggtagt ggtcggcgag ctgcacgccg ccgtcctcga tgttgtggcg
     1081 gatcttgaag ttggccttga tgccgttctt ctgcttgtcg gcggtgatat agacgttgtg
     1141 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     1201 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     1261 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
     1321 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcac
     1381 gccgtagccc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     1441 gatcagcttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     1501 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     1561 cagctcctcg cccttgctca ccatggtggc gaccggtagc gctagcttcg aaagatctgg
     1621 atcccagaag caaatgtaag caatagatgg ctctgccctg acttttatgc ccagcccgag
     1681 ctccgccggc gagaagctcc cagctctaca gatgttggaa tgagctggtt catgtgattg
     1741 ttgagttgaa caagctccca gatccatagc cgaagcttga ggaccaggtt ggctgacagt
     1801 gtggttccca tttaaaaggc accgcctatg ctatagttca tgtctgaagg cagggaaaat
     1861 ccagagtctc acctcaaata ctatcaggca gaagacattc tcctgtcctc aagccttttt
     1921 tttttttttt taaatatatg ttcaggcttt caacagattg catgaggcca cccactttag
     1981 cgagaacaat ctgcttactt tgtctactga ctcaatagtg aaactcttcc aaactccctc
     2041 atagacacac cccaaataat gtttgaacag aagtctgggc acctcatcat aaccatctgt
     2101 catggttaat ttcacatgcc accttgatga cctctagaat cacctgggca atgggtctcc
     2161 gggcatgtct gtagggaatg atcttgctag gttaactgag gttggaggac tcatccacgg
     2221 tgggtggcac cattccctag gctgggttct aggctatgta caaaagagga agcaaacgac
     2281 gcgtgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2341 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag
     2401 cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2461 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2521 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2581 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2641 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2701 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2761 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2821 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2881 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2941 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3001 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3061 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3121 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3181 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3241 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3301 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3361 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3421 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3481 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3541 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3601 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3661 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3721 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3781 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3841 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3901 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3961 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4021 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4081 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4141 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4201 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4261 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     4321 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     4381 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     4441 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac
     4501 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4561 tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt
     4621 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4681 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt
     4741 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4801 gcttgacggg gaaagccggc g
//
