LOCUS       sequence-411033-        4849 bp    DNA     circular SYN 22-APR-2025
DEFINITION  AiE0641m is an enhancer sequence designed to drive AAV-mediated 
            transgene expression in brain oligodendrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4849)
  AUTHORS   Mich JK, Sunil S, Johansen N, Martinez RA, Leytze M, Gore BB, 
            Mahoney JT, Ben-Simon Y, Bishaw Y, Brouner K, Campos J, Canfield R, 
            Casper T, Dee N, Egdorf T, Gary A, Gibson S, Goldy J, Groce EL, 
            Hirschstein D, Loftus L, Lusk N, Malone J, Martin NX, Monet D, 
            Omstead V, Opitz-Araya X, Oster A, Pom CA, Potekhina L, Reding M, 
            Rimorin C, Ruiz A, Sedeno-Cortes AE, Shapovalova NV, Taormina M, 
            Taskin N, Tieu M, Valera Cuevas NJ, Weed N, Way S, Yao Z, McMillen 
            DA, Kunst M, McGraw M, Thyagarajan B, Waters J, Bakken TE, Yao S, 
            Smith KA, Svoboda K, Podgorski K, Kojima Y, Horwitz GD, Zeng H, 
            Daigle TL, Lein ES, Tasic B, Ting JT, Levi BP
  TITLE     Enhancer-AAVs allow genetic access to oligodendrocytes and diverse 
            populations of astrocytes across species
  JOURNAL   bioRxiv 2023.09.20.558718
REFERENCE   2  (bases 1 to 4849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2023.09.20.558718"
COMMENT     Sequence Label: AiP12845 - pAAV-AiE0641m-minBG-SYFP2-WPRE3-BGH...
FEATURES             Location/Qualifiers
     source          1..4849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      861..870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(867..869,870..872,873..1583)
                     /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(912..933)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1173..1192)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1520..1541
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1676..1696)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    1883..1994
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2112..2252
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2112..2241
                     /label=AAV2 ITR
     rep_origin      2327..2782
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2414..2433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2624..2645
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2799..2818)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2918..2940
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2978..2996)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3064..3168
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3169..3237,3238..4029)
                     /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     complement(3387..3406)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4200..4788
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4689..4708
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagcat gaatggcctc tcaggatgga aggggggcgg
      181 ggagagaaga aagccggtca gaggcagcag ttgcttttaa gaatatgtaa gatttctagt
      241 ttctgaggtg ctgtgttata gcagctttaa gcaagaccta acaatttgtt aaatacttca
      301 tctccaaaca gtggtgggta ctacagtttc atgagcagca gataatagat tctgtttatt
      361 cgccaacaat accgacactg tttcgccccg gactgcaagt ggctgagcca tctggctcct
      421 atctccacag taaaacattg tccgtgacac aaaaaatgac tacatcccca ggacaaacta
      481 gtgtgacctc taaataaagc cttcctctga agctatgata tcacactgca caggggacac
      541 cagcacaatt gaatctggtc accaaaagcc caagtgagat aggactatgt aaatgggatt
      601 ttataggaga cgttttgtcc tctgtgcaaa ccatcgagat ccgtccacag cactgaaaca
      661 tctttttcat tcaccctgag gacaacagtg tgcttctttc tggggggttg gcagtcctct
      721 ttctccggcc ccaggccgag gaagcacaca tccgtgtccc agaggtggag ctcgggctgg
      781 gcataaaagt cagggcagag ccatctattg cttacatttg cttctgggat ccagatcttt
      841 cgaagctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
      901 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      961 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
     1021 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc gtgcagtgct
     1081 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1141 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1201 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1261 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1321 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     1381 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1441 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc
     1501 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1561 tcggcatgga cgagctgtac aagtaagtcg acggcgcgcc gcggccgcga attcgatatc
     1621 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     1681 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     1741 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     1801 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     1861 ccgtggctcg agagatcttc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1921 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1981 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2041 caggacagca agggggagga ttgggaagac aatagcaggc atgagatctc acgtgcggac
     2101 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2161 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     2221 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     2281 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     2341 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2401 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2461 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2521 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     2581 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2641 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     2701 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2761 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     2821 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2881 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2941 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3001 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3061 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3121 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3181 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3241 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3301 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3361 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3421 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3481 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3541 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3601 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3661 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3721 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3781 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3841 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3901 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3961 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4021 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4081 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4141 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4201 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4261 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4321 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4381 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4441 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4501 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4561 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4621 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4681 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4741 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4801 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
