LOCUS       sequence-194809-        8748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of SmyD1 and copGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8748)
  AUTHORS   Wei L, Murphy BL, Wu G, Parker M, Easton J, Gilbertson RJ, Zhang J, 
            Roussel MF
  TITLE     Exome sequencing analysis of murine medulloblastoma models 
            identifies WDR11 as a potential tumor suppressor in Group 3 tumors.
  JOURNAL   Oncotarget. 2017 Jul 27. doi: 10.18632/oncotarget.19642.
  PUBMED    28778067
REFERENCE   2  (bases 1 to 8748)
  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: "Oncotarget.
            2017 Jul 27. doi: 10.18632/oncotarget.19642."
COMMENT     Sequence Label: pCDH-EF1-mouse SmyD1-T2A-copGFP
FEATURES             Location/Qualifiers
     source          1..8748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     150..167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     151..167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        182..408
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             409..589
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    636..761
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1254..1487
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1672..1716
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1983..2099
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2142..2353
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2366..2634
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4166..4219
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4250..4912
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     misc_feature    5003..5591
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5056..5076)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5474..5485)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5665..5898
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5970..6091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6007..6026)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6061..6080
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6131..6266
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6299..6315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6299..6315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6312..6334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6323..6339
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6347..6353,6354..6371,6372..6377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6392..6413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6530..6547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6701..7289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7460..8251,8252..8320))
                     /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     8083..8102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8321..8425)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8493..8511
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8549..8571)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8671..8690
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
       61 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
      121 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgacgc
      181 gtgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
      241 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
      301 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
      361 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
      421 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      481 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      541 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      601 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      661 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
      721 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
      781 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
      841 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
      901 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
      961 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1021 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1081 agaccaccgc acagcaagcg gccactgatc ttcagacctg gaggaggaga tatgagggac
     1141 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1201 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1261 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1321 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1381 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1441 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1501 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1561 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1621 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1681 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     1741 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     1801 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     1861 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     1921 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta
     1981 acttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     2041 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaatt caaaatttta
     2101 tcgatactag tggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc
     2161 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg
     2221 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     2281 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     2341 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc
     2401 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
     2461 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
     2521 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
     2581 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
     2641 gtgaccggcg cctactctag acctcgaagg cctgacagac tctgagatga caataggcag
     2701 catggagaac gtggaggtct tcacttccga gggcaaaggc agaggtctga aggctacgaa
     2761 ggagttctgg gctgcggatg tcatctttgc ggagagggct tattctgcag tggtttttga
     2821 cagcctcatt aacttcgtgt gccacacctg cttcaagagg caggagaagc tccatcgctg
     2881 cgggcagtgc aagttcgccc attactgcga ccgcacgtgc cagaaggatg cttggctgaa
     2941 tcacaagaac gagtgcgctg ccatcaagaa atatgggaaa gtgcccaacg agaacatcag
     3001 gctggccgcc cgcatcatgt ggcgggtgga gagagagggc actgggctca cagagggctg
     3061 cctggtgtcc gtggatgact tacagaacca cgtggagcac tttggggagg aggagcagaa
     3121 ggaactccga gtagacgtgg acacgttctt gcagtactgg ccaccacaga gccagcagtt
     3181 cagcatgcag tatatctcac acatctttgg tgtgatcaac tgcaacggtt tcactctcag
     3241 tgaccagaga gggctacagg cagtaggtgt gggcatcttc cccaacctgg gcctggtgaa
     3301 ccatgattgc tggccaaact gcactgtcat attcaacaat ggcaatcatg aggcagtgaa
     3361 atccatgttt cacacgcaga tgaggattga gctccgggcc ctgggcaaga tctcagaagg
     3421 cgaggagctg actgtgtcct acatagactt cctgcacctc agtgaggagc gcaggcggca
     3481 gctgaagaaa cagtactact ttgactgctc ctgtgagcac tgccagaagg ggctgaagga
     3541 cgacctcttc ctggcagcga aggaagaccc caagccctcc caggaagtgg tgaaggagat
     3601 gatacaattc tcaaaggata cactggaaaa aatagacaag gctcgctccg agggtttgta
     3661 tcacgaggtt gtgaagctgt gtcgggagtg cctggagaag caggagccag tgttcgccga
     3721 caccaacctc tacgtgcttc ggctgctcag cattgcatca gaggtcctct cctacctcca
     3781 ggcctatgag gaggcctcac attatgccag gaggatggtt gatggctaca tgaaactcta
     3841 ccaccataac aatgcccaac tgggcatggc tgtgatgagg gcagggctta ccaactggca
     3901 tgctggtcac atcgaagtgg ggcatgggat gatctgcaaa gcctatgcta ttctcctggt
     3961 gacccatgga ccctcccacc ctatcaccaa agacttagag gccatgcgga tgcagacaga
     4021 gatggagctg cgtatgttcc gccaaaacga attcatgtat cacaagatgc gagaggctgc
     4081 cctgaacaac cagcccatgc aggtcatggc tgagcctagc aatgaaccag cccccgctct
     4141 gttccataag aagcaggcgg ccgctgaggg cagaggaagt cttctaacat gcggtgacgt
     4201 ggaggagaat cccggccctt ccggaatgga gagcgacgag agcggcctgc ccgccatgga
     4261 gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg
     4321 agagggcacc cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct
     4381 gaccttcagc ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac
     4441 ctaccccagc ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa
     4501 cacccgcatc gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta
     4561 cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag
     4621 cgtgatcttc accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat
     4681 gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta
     4741 ctacagcttc gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct
     4801 gcagaacggg ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga
     4861 gctgggcatc gtggagtacc agcacgcctt caagaccccc atcgccttcg ccagatcccg
     4921 cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg
     4981 ctagagctga atctaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     5041 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5101 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5161 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5221 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5281 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5341 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     5401 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5461 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5521 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5581 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5641 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agataagatc
     5701 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5761 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5821 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5881 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     5941 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     6001 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6061 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     6121 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6181 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6241 gtagtgagga ggcttttttg gaggcctaga cttttgcaga gaccaaattc gtaatcatgt
     6301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7201 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7261 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7321 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7381 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7441 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7501 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7561 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7621 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7681 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7741 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7801 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7861 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7921 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7981 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8041 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8101 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8161 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8221 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8281 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8341 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8401 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     8461 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     8521 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     8581 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     8641 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     8701 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaatacc
//
