LOCUS       sequence-238086-        7262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse Wnt5a in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7262)
  AUTHORS   Rodriguez JP, Coulter M, Miotke J, Meyer RL, Takemaru K, Levine JM
  TITLE     Abrogation of beta-catenin signaling in oligodendrocyte precursor 
            cells reduces glial scarring and promotes axon regeneration after 
            CNS injury.
  JOURNAL   J Neurosci. 2014 Jul 30;34(31):10285-97. doi: 
            10.1523/JNEUROSCI.4915-13.2014.
  PUBMED    25080590
REFERENCE   2  (bases 1 to 7262)
  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"; doi: 
            "10.1523/JNEUROSCI.4915-13"; journalName: "J Neurosci"; date: 
            "2014-07-30- 30"; volume: "34"; issue: "31"; pages: "10285-97"
COMMENT     Sequence Label: pAAV-IRES-hrGFP-mWnt5a
FEATURES             Location/Qualifiers
     source          1..7262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          353..731
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     promoter        802..820
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2033..2104
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    join(2140..2711,2712..2714,2715..2723)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2307..2324)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2534..2553
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2724..3443
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     complement(3473..3492)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3474..3492)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3500..3976
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3699..3718)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4016..4156
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4016..4145
                     /label=AAV2 ITR
     rep_origin      4231..4686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4318..4337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4528..4549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4703..4722)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4822..4844
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4882..4900)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4968..5072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5073..5141,5142..5933)
                     /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(5291..5310)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6104..6692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6593..6612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6754..6883
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        6959..7262
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggattcgaa
      301 tcccggccgg gaacggtgca ttggaacgcg gattccccgt gccaagagtg acgtaagtac
      361 cgcctataga gtctataggc ccacaaaaaa tgctttcttc ttttaatata cttttttgtt
      421 tatcttattt ctaatacttt ccctaatctc tttctttcag ggcaataatg atacaatgta
      481 tcatgcctct ttgcaccatt ctaaagaata acagtgataa tttctgggtt aaggcaatag
      541 caatatttct gcatataaat atttctgcat ataaattgta actgatgtaa gaggtttcat
      601 attgctaata gcagctacaa tccagctacc attctgcttt tattttatgg ttgggataag
      661 gctggattat tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct
      721 tcctcccaca gctcctgggc aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag
      781 aattgggatt cgaacatcga taattaaccc tcactaaagg gaacaaaagc tggagctcca
      841 ccgcggtggc ggccgctcta gcccgggcgg atccgaattc accatgaaga agcccattgg
      901 aatattaagc ccgggagtgg ctttggggac cgctggaggt gccatgtctt ccaagttctt
      961 cctaatggct ttggccacgt ttttctcctt cgcccaggtt gttatagaag ctaattcttg
     1021 gtggtctcta ggtatgaata accctgttca gatgtcagaa gtatatatca taggtgcaca
     1081 gcctctctgc agccaactgg caggactttc tcaaggacag aagaaactct gccacttgta
     1141 tcaggaccac atgcagtaca ttggagaagg tgcgaagaca ggcatcaagg aatgccagta
     1201 ccagttccgg catcggagat ggaactgcag cacagtggac aatacttctg tctttggcag
     1261 ggtgatgcaa ataggcagcc gagagacggc cttcacgtac gcggtgagcg cagctggggt
     1321 ggtgaacgcc atgagccgag catgccggga gggcgagctg tctacctgtg gctgcagccg
     1381 cgctgcgcgc cccaaggacc tgcctcggga ctggttgtgg ggcggctgcg gagacaacat
     1441 cgactatggc taccgcttcg ccaaggagtt cgtggacgct agagaaaggg aacgaatcca
     1501 cgctaagggt tcctatgaga gcgcacgcat cctcatgaac ttacacaaca atgaagcagg
     1561 ccgtaggaca gtatacaacc tggcagatgt agcctgtaag tgtcatggag tgtctggctc
     1621 ctgtagcctc aagacgtgct ggctgcagct ggcggacttc cggaaggtgg gcgatgccct
     1681 caaggagaag tatgatagcg cggcggccat gaggctcaac agccggggca agctggtgca
     1741 ggtcaacagc cgcttcaact ccccgaccac gcaggacctg gtctacatcg accccagtcc
     1801 ggactactgt gtgcgcaacg agagcactgg ctcgctgggc acgcagggac gcctgtgcaa
     1861 caagacctca gaggggatgg acggctgcga gctcatgtgc tgtgggcgtg gctatgacca
     1921 gtttaagaca gtgcagaccg aacgctgtca ttgcaagttt cactggtgct gctatgtcaa
     1981 atgcaagaag tgcacggaga ttgtggatca gttcgtgtgc aaatagctcg aggactacaa
     2041 ggatgacgat gacaaggatt acaaagacga cgatgataag gactataagg atgatgacga
     2101 caaataatag caattcctcg acgactgcat agggttaccc ccctctccct cccccccccc
     2161 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     2221 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     2281 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     2341 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     2401 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2461 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     2521 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     2581 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     2641 gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     2701 acacgatgat aatggccaca accatggtga gcaagcagat cctgaagaac accggcctgc
     2761 aggagatcat gagcttcaag gtgaacctgg agggcgtggt gaacaaccac gtgttcacca
     2821 tggagggctg cggcaagggc aacatcctgt tcggcaacca gctggtgcag atccgcgtga
     2881 ccaagggcgc ccccctgccc ttcgccttcg acatcctgag ccccgccttc cagtacggca
     2941 accgcacctt caccaagtac cccgaggaca tcagcgactt cttcatccag agcttccccg
     3001 ccggcttcgt gtacgagcgc accctgcgct acgaggacgg cggcctggtg gagatccgca
     3061 gcgacatcaa cctgatcgag gagatgttcg tgtaccgcgt ggagtacaag ggccgcaact
     3121 tccccaacga cggccccgtg atgaagaaga ccatcaccgg cctgcagccc agcttcgagg
     3181 tggtgtacat gaacgacggc gtgctggtgg gccaggtgat cctggtgtac cgcctgaaca
     3241 gcggcaagtt ctacagctgc cacatgcgca ccctgatgaa gagcaagggc gtggtgaagg
     3301 acttccccga gtaccacttc atccagcacc gcctggagaa gacctacgtg gaggacggcg
     3361 gcttcgtgga gcagcacgag accgccatcg cccagctgac cagcctgggc aagcccctgg
     3421 gcagcctgca cgagtgggtg taatagggta ccaggtaagt gtacccaatt cgccctatag
     3481 tgagtcgtat tagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     3541 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     3601 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     3661 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     3721 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     3781 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     3841 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     3901 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     3961 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     4021 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4081 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4141 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4201 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4261 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4321 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4381 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4441 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4501 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4561 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4621 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4681 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4741 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4801 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4861 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4921 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4981 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5041 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5101 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5161 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5221 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5281 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5341 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5401 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5461 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5521 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5581 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5641 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5701 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5761 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5821 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5881 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5941 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6001 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6061 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6241 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6721 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc
     6781 tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     6841 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg cacgcgtcta
     6901 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     6961 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     7021 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     7081 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     7141 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     7201 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     7261 tg
//
