LOCUS       sequence-214942-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7073)
  AUTHORS   Lindsey JC, Kawauchi D, Schwalbe EC, Solecki DJ, Selby MP, McKinnon 
            PJ, Olson JM, Hayden JT, Grundy RG, Ellison DW, Williamson D, Bailey
            S, Roussel MF, Clifford SC
  TITLE     Cross-species epigenetics identifies a critical role for VAV1 in SHH
            subgroup medulloblastoma maintenance.
  JOURNAL   Oncogene. 2015 Sep 3;34(36):4746-57. doi: 10.1038/onc.2014.405. Epub
            2014 Dec 22.
  PUBMED    25531316
REFERENCE   2  (bases 1 to 7073)
  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: 
            "10.1038/onc.2014.405"; journalName: "Oncogene"; date: "2015-09-3- 
            3"; volume: "34"; issue: "36"; pages: "4746-57"
COMMENT     Sequence Label: MSCV mouse Vav1DN IRES GFP
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             171..687
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    751..1092
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1159..1575
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1505..1527
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1543..1559
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1607..1616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2205..2754
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2349..2366)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2576..2595
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2757..2759,2760..2762,2763..3476)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2757..2759,2760..2762,2763..3473)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2802..2823)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3063..3082)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3410..3431
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3638..4152
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4314..4330
                     /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(4338..4344,4345..4362,4363..4368))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4383..4404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4521..4538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4692..5280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4772..4791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5451..6242,6243..6311))
                     /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     6074..6093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6312..6416)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6484..6502
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6540..6562)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6662..6681
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6875..6897
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7025..7044
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata
       61 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac
      121 gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
      181 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata
      241 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg
      301 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
      361 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
      421 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
      481 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
      541 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct
      601 tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct cagattgatt
      661 gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga gacccctgcc
      721 cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc
      781 tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct gtactagtta
      841 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc
      901 cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga
      961 agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac
     1021 ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg
     1081 tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg
     1141 tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac
     1201 tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg
     1261 gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc
     1321 tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga
     1381 cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc
     1441 tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct
     1501 ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact
     1561 ccttctctag gcgccggaat tgggatccta aagtgagagg ccggcagcca ccatggagct
     1621 ctggcgacag tgcacccact ggctgatcca gtgtcgggtg ctgcctccca gccaccgtgt
     1681 gacctgggag ggggcccagg tgtgtgagct ggcacaggca ctgcgggacg gtgtcctctt
     1741 gtgccaattg cttaacaacc tgcttcccca ggccattaat cttcgcgagg ttaacttgcg
     1801 gccccagatg tcccagttcc tttgtcttaa gaacattcga accttcctgt ctacttgctg
     1861 tgagaagttc ggcctcaagc gcagtgaact ctttgaggct tttgacctct tcgatgtgca
     1921 ggactttgga aaggtcatct acaccctgtc tgctctgtca tggacaccca ttgcccagaa
     1981 caaaggaatc atgcccttcc caacagagga cagcgctctg aacgacgaag atatttacag
     2041 tggcctttca gaccagattg atgacaccgc agaggaagac gaggaccttt atgactgcgt
     2101 ggaaaatgag gaggcagagg gggacgagat ctacgaggac ctaatgcgct tggagtcggt
     2161 gcctacgcca tgactcgagc gggatcaatt ccgccccccc cctaacgtta ctggccgaag
     2221 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     2281 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     2341 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     2401 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     2461 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     2521 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     2581 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     2641 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     2701 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat aataccatgg
     2761 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2821 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2881 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2941 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     3001 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     3061 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3121 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3181 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3241 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3301 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3361 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3421 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     3481 gccgcgactc tagagtcgac ctgcaggcat gcaagcttca ggtagccggc taacgttaac
     3541 aaccggtacc tctagaacta tagctagcat gcgcaaattt aaagcgctga tatcgataaa
     3601 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     3661 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata
     3721 gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat
     3781 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     3841 cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     3901 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     3961 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
     4021 cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     4081 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     4141 ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt
     4201 cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact
     4261 cctgaaatag ttcattatgg acagcgcaga aagagctggg gagaattgtg aaattgttat
     4321 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4381 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4441 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4501 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4561 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4621 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4681 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4741 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4801 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4861 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4921 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4981 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5041 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5101 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5161 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5221 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5281 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5341 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5401 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5461 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5521 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5581 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5641 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5701 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5761 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5821 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5881 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5941 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6001 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6061 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6121 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6181 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6241 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6301 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6361 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6421 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     6481 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     6541 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     6601 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     6661 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     6721 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     6781 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     6841 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     6901 gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     6961 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     7021 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtg
//
