pAAV-IRES-hrGFP-mWnt3a

LOCUS      pAAV-IRES-hrGFP-mWnt3a               7177 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses mouse Wnt3a in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-IRES-hrGFP-mWnt3a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7177)
  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 7177)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7177
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1106..1124
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2253..2324
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     misc_feature    2360..2942
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2526..2543)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2753..2772
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2943..3662
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     primer_bind     complement(3692..3711)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3693..3711)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3719..4195
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3918..3937)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4235..4375
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4235..4364
                     /label=AAV2 ITR
     rep_origin      4450..4905
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4537..4556)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4747..4768
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4922..4941)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5041..5063
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5101..5119)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5187..5291
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5292..6152
                     /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(5510..5529)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6323..6911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6812..6831
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6973..7102
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggatt
      601 cgaatcccgg ccgggaacgg tgcattggaa cgcggattcc ccgtgccaag agtgacgtaa
      661 gtaccgccta tagagtctat aggcccacaa aaaatgcttt cttcttttaa tatacttttt
      721 tgtttatctt atttctaata ctttccctaa tctctttctt tcagggcaat aatgatacaa
      781 tgtatcatgc ctctttgcac cattctaaag aataacagtg ataatttctg ggttaaggca
      841 atagcaatat ttctgcatat aaatatttct gcatataaat tgtaactgat gtaagaggtt
      901 tcatattgct aatagcagct acaatccagc taccattctg cttttatttt atggttggga
      961 taaggctgga ttattctgag tccaagctag gcccttttgc taatcatgtt catacctctt
     1021 atcttcctcc cacagctcct gggcaacgtg ctggtctgtg tgctggccca tcactttggc
     1081 aaagaattgg gattcgaaca tcgataatta accctcacta aagggaacaa aagctggagc
     1141 tccaccgcgg tggcggccgc tctagcccgg gcggatccga attcaccatg gctcctctcg
     1201 gatacctctt agtgctctgc agcctgaagc aggctctggg cagctacccg atctggtggt
     1261 ccttggctgt gggaccccag tactcctctc tgagcactca gcccattctc tgtgccagca
     1321 tcccaggcct ggtaccgaag cagctgcgct tctgcaggaa ctacgtggag atcatgccca
     1381 gcgtggctga gggtgtcaaa gcgggcatcc aggagtgcca gcaccagttc cgaggccggc
     1441 gttggaactg caccaccgtc agcaacagcc tggccatctt tggccctgtt ctggacaaag
     1501 ccacccggga gtcagccttt gtccatgcca tcgcctccgc tggagtagct ttcgcagtga
     1561 cacgctcctg tgcagaggga tcagctgcta tctgtgggtg cagcagccgc ctccagggct
     1621 ccccaggcga gggctggaag tggggcggct gtagtgagga cattgaattt ggaggaatgg
     1681 tctctcggga gtttgccgat gccagggaga accggccgga tgcccgctct gccatgaacc
     1741 gtcacaacaa tgaggctggg cgccaggcca tcgccagtca catgcacctc aagtgcaaat
     1801 gccacgggct atctggcagc tgtgaagtga agacctgctg gtggtcgcag ccggacttcc
     1861 gcaccatcgg ggatttcctc aaggacaagt atgacagtgc ctcggagatg gtggtagaga
     1921 aacaccgaga gtctcgtggc tgggtggaga ccctgaggcc acgttacacg tacttcaagg
     1981 tgccgacaga acgcgacctg gtctactacg aggcctcacc caacttctgc gaacctaacc
     2041 ccgaaaccgg ctccttcggg acgcgtgacc gcacctgcaa tgtgagctcg catggcatag
     2101 atgggtgcga cctgttgtgc tgcgggcgcg ggcataacgc gcgcactgag cgacggaggg
     2161 agaaatgcca ctgtgttttc cattggtgct gctacgtcag ctgccaggag tgcacacgtg
     2221 tctatgacgt gcacacctgc aagtagctcg aggactacaa ggatgacgat gacaaggatt
     2281 acaaagacga cgatgataag gactataagg atgatgacga caaataatag caattcctcg
     2341 acgactgcat agggttaccc ccctctccct ccccccccnt aacgttactg gccgaagccg
     2401 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     2461 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     2521 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     2581 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     2641 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     2701 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     2761 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     2821 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaacgtcta
     2881 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atggccacaa
     2941 ccatggtgag caagcagatc ctgaagaaca ccggcctgca ggagatcatg agcttcaagg
     3001 tgaacctgga gggcgtggtg aacaaccacg tgttcaccat ggagggctgc ggcaagggca
     3061 acatcctgtt cggcaaccag ctggtgcaga tccgcgtgac caagggcgcc cccctgccct
     3121 tcgccttcga catcctgagc cccgccttcc agtacggcaa ccgcaccttc accaagtacc
     3181 ccgaggacat cagcgacttc ttcatccaga gcttccccgc cggcttcgtg tacgagcgca
     3241 ccctgcgcta cgaggacggc ggcctggtgg agatccgcag cgacatcaac ctgatcgagg
     3301 agatgttcgt gtaccgcgtg gagtacaagg gccgcaactt ccccaacgac ggccccgtga
     3361 tgaagaagac catcaccggc ctgcagccca gcttcgaggt ggtgtacatg aacgacggcg
     3421 tgctggtggg ccaggtgatc ctggtgtacc gcctgaacag cggcaagttc tacagctgcc
     3481 acatgcgcac cctgatgaag agcaagggcg tggtgaagga cttccccgag taccacttca
     3541 tccagcaccg cctggagaag acctacgtgg aggacggcgg cttcgtggag cagcacgaga
     3601 ccgccatcgc ccagctgacc agcctgggca agcccctggg cagcctgcac gagtgggtgt
     3661 aatagggtac caggtaagtg tacccaattc gccctatagt gagtcgtatt agatctacgg
     3721 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     3781 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     3841 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     3901 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     3961 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     4021 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     4081 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     4141 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     4201 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     4261 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4321 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     4381 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4441 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4501 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     4561 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4621 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4681 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4741 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     4801 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     4861 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4921 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4981 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5041 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5101 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5161 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5221 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5281 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5341 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5401 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5461 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5521 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5581 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5641 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5701 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5761 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5821 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5881 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5941 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6001 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6061 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6121 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6181 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6241 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6301 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6361 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6421 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6481 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6541 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6601 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6661 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6721 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6781 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6841 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6901 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6961 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
     7021 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
     7081 aactccatca ctaggggttc ctgcggccgc acgcgtctag ttattaatag taatcaatta
     7141 cggggtcatt agttcatagc ccatatatgg agttccg
//