LOCUS       sequence-378610-        6200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses membrane-targeted EGFP in oligodendrocytes; AAV vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6200)
  AUTHORS   Lam M, Takeo K, Almeida RG, Cooper MH, Wu K, Iyer M, Kantarci H, 
            Zuchero JB
  TITLE     CNS myelination requires VAMP2/3-mediated membrane expansion in 
            oligodendrocytes.
  JOURNAL   Nat Commun. 2022 Sep 23;13(1):5583. doi: 10.1038/s41467-022-33200-4.
  PUBMED    36151203
REFERENCE   2  (bases 1 to 6200)
  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: "Nat 
            Commun."; date: "2022-09-23"; volume: "13(1):5583. doi"; pages: " 
            10.1038/s41467-022-33200-4"
COMMENT     Sequence Label: AAV_pMBP-EGFP-caax
FEATURES             Location/Qualifiers
     source          1..6200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   20..149
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2154..2163
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2160..2162,2163..2165,2166..2876)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2160..2162,2163..2165,2166..2876)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2205..2226)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2466..2485)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2813..2834
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2966..3442
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3165..3184)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3482..3622
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3482..3611
                     /label=AAV2 ITR
     rep_origin      3697..4152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3784..3803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3994..4015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4169..4188)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4288..4310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4348..4366)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4434..4538
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4539..4607,4608..5399)
                     /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(4757..4776)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5570..6158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6059..6078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
       61 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      121 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtgagctcc ttcctgctta
      181 ggccgtgtct gctctgtcag tctttcctct ggctctagtc tgtccatctg cagggataac
      241 ctcccaaggg ccctcttgtt ggctgttcac cttccgtctg tcttcatagc ctcctgatga
      301 gagagcgcgc acacacaggc tacctgactg atgtgttgct gcactgtggg gtccagtgac
      361 acagctgtgg aggtcaggaa cccagaatta agctgtctgg gagccacaag tttcaagagt
      421 ccatgctatg gctcctacat gcaggtggtg ccagcagcat tccttggtgt cctgtggctg
      481 tggttgcatc tctgactctc ctccacctat cacattcctt ctccagtacc aggcctctgt
      541 gtctcagttt tcctctctca caagatccct agcttcagga ttgcacccac ctccactagc
      601 catgagcccg tcctcactgg atcacgtctg caaagactct atttccagtt aaagtcacag
      661 acacgggcat agagaggggg ataggatatg aaaatatcag ttgggggagg ggcattcaat
      721 tcttagctca cattttcttc atccaatttc ccccactaga atgtaagctt tgagagaaaa
      781 gggaccagat cttattcctc accgtggctt taacacttag agaaaatgca tcccctctaa
      841 tcaataagtc atcgacagtg ggtagatgga ggaacggcag tgcgtagtag gatgcgtgca
      901 agcatagtct cgtgcatggg tgcatagatc gctgggcagg tggacaaggt gggggtggat
      961 aaagaagtgg gtagatgatt gatgttaggt aaatatcact gggtggacag atgggtggta
     1021 ggtggatgga tggttagaat agtcagaaga gggatggatt gataaggtga acagatgata
     1081 aatgggtgat agactggaag ggttgtcaaa agaggataag ggaagtgtga gctagccgta
     1141 tttctaaggt cagtaataga gttgggagaa gaggttaagt tacatccatt taaacctcac
     1201 acgaagctga gagggaatgg acttgctgcc gttggtgagg aaagcgttgc atttcccgtg
     1261 tgcttggttg tgaagtgctc aggtcccaca tgaagcagtc aggttactgc ggcttacaga
     1321 ggagccagat ccaaatgccc cgagtaagca cgtccccgag ccagaggcct ccagcggaat
     1381 ccgggagagg gattgctcag tgccctgctt ccctggactg taagctgcag aaagatgtgg
     1441 gaagtcctgt tctccactga gaacactaaa agcacctttt gtcaaacgac cgcttcacat
     1501 ctggggcttg tgcactggtg gccttttaaa ccagagacaa cccacaagat acctaacctg
     1561 cggggctctc tggtacagtg agcaactcag gaaatgcttt ggcttgattg ctgtgggctc
     1621 tcaggccatc gccctctgga gtggttcttt taatgagaac ctgaagattg gcccctgagc
     1681 catgtatacc aagcaagctc aatccaggtt agctccctct ggttggggca agctaacgtg
     1741 ctccttgggc cccgcgcgta actgtgcgtt ttataggaga cagctagttc aagaccccag
     1801 gaagaaagcg gctttgtccc cctctaggcc tcgtacaggc ccacattcat atctcattgt
     1861 tgttgcaggg gaggcagatg cgatccagaa caatgggacc tcggctgagg acacggcggt
     1921 gacagactcc aagcacacag cagacccaaa gaataactgg caaggcgccc acccagctga
     1981 cccagggaac cgcccccact tgatccgcct cttttcccga gatgccccgg gaagggagga
     2041 caacaccttc aaagacaggc cctcagagtc cgacgagctt cagaccatcc aagaagaccc
     2101 cacagcagct tccggaagct cgaataattc taggatccct agcgctaccg gtcgccacca
     2161 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2221 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2281 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2341 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2401 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2461 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2521 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2581 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2641 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2701 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2761 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2821 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg
     2881 gactcagatc taagctgaac cctcctgatg agagtggccc cggctgcatg agctgcaagt
     2941 gtgtgctctc ctgataaaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     3001 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     3061 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     3121 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     3181 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     3241 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     3301 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     3361 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3421 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     3481 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3541 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3601 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3661 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     3721 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     3781 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3841 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3901 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3961 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4021 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4081 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4141 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4201 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4261 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4321 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4381 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4501 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4561 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4621 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4681 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4741 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4801 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4861 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4921 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4981 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5041 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5101 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5161 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5221 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5281 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5341 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5401 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5461 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5521 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5701 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5761 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5821 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5881 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5941 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6001 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6061 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6121 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6181 tacggttcct ggccttttgc
//
