LOCUS       sequence-411042-        6663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct-expressing SYFP2 in oligodendrocytes AAV Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6663)
  TITLE     Allen Institute Plasmids Aug 2023
REFERENCE   2  (bases 1 to 6663)
  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"
COMMENT     Sequence Label: AiP14084 -
            pAAV-mMbp_promoter-SYFP2-P2A-3XFLAG...
FEATURES             Location/Qualifiers
     source          1..6663
                     /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      2129..2138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2135..2137,2138..2140,2141..2851)
                     /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(2441..2460)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2788..2809
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2861..2917
                     /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             2930..2995
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3026..3403
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="PEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVL
                     KQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPG
                     ELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(3490..3510)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3697..3808
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3926..4066
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3926..4055
                     /label=AAV2 ITR
     rep_origin      4141..4596
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4228..4247)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4438..4459
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4613..4632)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4732..4754
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4792..4810)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4878..4982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4983..5051,5052..5843)
                     /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(5201..5220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6014..6602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6503..6522
                     /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 gcgttccttc ctgcttaggc cgtgtctgct ctgtcagtct
      181 ttcctctggc tctagtctgt ccatctgcag ggataacctc ccaagggccc tcttgttggc
      241 tgttcacctt ccgtctgtct tcatagcctc ctgatgagag agcgcgcaca cacaggctac
      301 ctgactgatg tgttgctgca ctgtggggtc cagtgacaca gctgtggagg tcaggaaccc
      361 agaattaagc tgtctgggag ccacaagttt caagagtcca tgctatggct cctacatgca
      421 ggtggtgcca gcagcattcc ttggtgtcct gtggctgtgg ttgcatctct gactctcctc
      481 cacctatcac attccttctc cagtaccagg cctctgtgtc tcagttttcc tctctcacaa
      541 gatccctagc ttcaggattg cacccacctc cactagccat gagcccgtcc tcactggatc
      601 acgtctgcaa agactctatt tccagttaaa gtcacagaca cgggcataga gagggggata
      661 ggatatgaaa atatcagttg ggggaggggc attcaattct tagctcacat tttcttcatc
      721 caatttcccc cactagaatg taagctttga gagaaaaggg accagatctt attcctcacc
      781 gtggctttaa cacttagaga aaatgcatcc cctctaatca ataagtcatc gacagtgggt
      841 agatggagga acggcagtgc gtagtaggat gcgtgcaagc atagtctcgt gcatgggtgc
      901 atagatcgct gggcaggtgg acaaggtggg ggtggataaa gaagtgggta gatgattgat
      961 gttaggtaaa tatcactggg tggacagatg ggtggtaggt ggatggatgg ttagaatagt
     1021 cagaagaggg atggattgat aaggtgaaca gatgataaat gggtgataga ctggaagggt
     1081 tgtcaaaaga ggataaggga agtgtgagct agccgtattt ctaaggtcag taatagagtt
     1141 gggagaagag gttaagttac atccatttaa acctcacacg aagctgagag ggaatggact
     1201 tgctgccgtt ggtgaggaaa gcgttgcatt tcccgtgtgc ttggttgtga agtgctcagg
     1261 tcccacatga agcagtcagg ttactgcggc ttacagagga gccagatcca aatgccccga
     1321 gtaagcacgt ccccgagcca gaggcctcca gcggaatccg ggagagggat tgctcagtgc
     1381 cctgcttccc tggactgtaa gctgcagaaa gatgtgggaa gtcctgttct ccactgagaa
     1441 cactaaaagc accttttgtc aaacgaccgc ttcacatctg gggcttgtgc actggtggcc
     1501 ttttaaacca gagacaaccc acaagatacc taacctgcgg ggctctctgg tacagtgagc
     1561 aactcaggaa atgctttggc ttgattgctg tgggctctca ggccatcgcc ctctggagtg
     1621 gttcttttaa tgagaacctg aagattggcc cctgagccat gtataccaag caagctcaat
     1681 ccaggttagc tccctctggt tggggcaagc taacgtgctc cttgggcccc gcgcgtaact
     1741 gtgcgtttta taggagacag ctagttcaag accccaggaa gaaagcggct ttgtccccct
     1801 ctaggcctcg tacaggccca cattcatatc tcattgttgt tgcaggggag gcagatgcga
     1861 tccagaacaa tgggacctcg gctgaggaca cggcggtgac agactccaag cacacagcag
     1921 acccaaagaa taactggcaa ggcgcccacc cagctgaccc agggaaccgc ccccacttga
     1981 tccgcctctt ttcccgagat gccccgggaa gggaggacaa caccttcaaa gacaggccct
     2041 cagagtccga cgagcttcag accatccaag aagaccccac agcagcttcc ggagggatcc
     2101 agatctttcg aagctagcgc taccggtcgc caccatggtc tccaagggcg aggagctgtt
     2161 caccggggtg gtgcccatcc tggtcgagct ggacggcgat gtcaacggcc acaagttcag
     2221 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
     2281 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcgt
     2341 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2401 gcccgaaggc tacgtccagg agcgcaccat cttcttcaaa gacgacggca actacaagac
     2461 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2521 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2581 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     2641 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     2701 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaagctgag
     2761 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2821 gatcactctc ggcatggacg agctgtacaa gggctctggt gctaccaact tctcactgtt
     2881 gaaacaggca ggggatgtag aggagaatcc agggcctggt gctagtggag actacaaaga
     2941 ccatgacgga gattataaag atcatgacat cgattacaag gatgacgatg acaagtccgg
     3001 actcagatct ggaggctccg gaggcccaga gccagcgaag tctgctcccg ccccgaaaaa
     3061 gggctccaag aaggcggtga ctaaggcgca gaagaaaggc ggcaagaagc gcaagcgcag
     3121 ccgcaaggag agctattcca tctatgtgta taaggttctg aagcaggtcc accctgacac
     3181 cggcatttcg tccaaggcca tgggcatcat gaactcgttt gtgaacgaca ttttcgagcg
     3241 catcgcaggt gaggcttccc gcctggcgca ttacaacaag cgctcgacca tcacctccag
     3301 ggagatccag acggccgtgc gcctgctgct gcctggggag ttggccaagc acgccgtgtc
     3361 cgagggtact aaggccatca ccaagtacac cagcgctaag taatgaggcg cgccgcggcc
     3421 gcgaattcga tatcataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3481 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3541 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct
     3601 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     3661 gggcactgac aattccgtgg ctcgagagat cttcgactgt gccttctagt tgccagccat
     3721 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3781 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3841 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgaga
     3901 tctcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3961 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4021 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4081 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4141 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4201 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4261 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4321 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4381 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4441 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4501 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4561 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4621 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4681 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4741 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4801 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4861 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4981 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5041 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5101 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5161 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5221 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5281 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5341 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5401 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5461 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5521 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5581 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5641 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5701 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5761 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5821 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6661 tgt
//
