LOCUS       sequence-181580-        7799 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Retroviral expression of mouse Gfi1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7799)
  AUTHORS   Vo BT, Li C, Morgan MA, Theurillat I, Finkelstein D, Wright S, Hyle 
            J, Smith SM, Fan Y, Wang YD, Wu G, Orr BA, Northcott PA, Shilatifard
            A, Sherr CJ, Roussel MF
  TITLE     Inactivation of Ezh2 Upregulates Gfi1 and Drives Aggressive 
            Myc-Driven Group 3 Medulloblastoma.
  JOURNAL   Cell Rep. 2017 Mar 21;18(12):2907-2917. doi: 
            10.1016/j.celrep.2017.02.073.
  PUBMED    28329683
REFERENCE   2  (bases 1 to 7799)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2017.02"; journalName: "Cell Rep"; date: 
            "2017-03-21- 21"; volume: "18"; issue: "12"; pages: "2907-2917"
COMMENT     Sequence Label: MSCV-mouse Gfi1-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..7799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             140..656
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    720..1061
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1128..1544
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1474..1496
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1512..1528
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2900..3449
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3044..3061)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3271..3290
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3452..3454,3455..3457,3458..4171)
                     /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(3452..3454,3455..3457,3458..4168)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3497..3518)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3758..3777)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4105..4126
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4333..4847
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5009..5025
                     /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(5033..5039,5040..5057,5058..5063))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5078..5099
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5216..5233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5387..5975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5467..5486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6146..6937,6938..7006))
                     /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     6769..6788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7007..7111)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7179..7197
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7235..7257)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7357..7376
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7570..7592
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7720..7739
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac
       61 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
      121 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
      181 taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag
      241 gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc
      301 tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt
      361 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
      421 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
      481 caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc
      541 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg
      601 atccttggga gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga
      661 ggttccaccg agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta
      721 agctggccag cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct
      781 aatgtttgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
      841 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
      901 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
      961 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     1021 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     1081 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     1141 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     1201 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     1261 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     1321 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     1381 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     1441 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     1501 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt gggatcccag
     1561 tgtggtggta cgtaggaatt cgatatcacc ggtaccatgc cgcgctcatt cctggtcaag
     1621 agcaagaagg cgcacagcta tcaccagccg cgttctccgg ggccggacta ctccctgcgc
     1681 ctggagaccg tgcctgcgcc gggcagagca gagggcggcg ctgtgagtgc aggcgagtcg
     1741 aaaatggagc cccgagagcg tttgtccccc gactctcagc ttaccgaggc tcccgacagg
     1801 gcctccgcgt cccccaacag ctgcgaaggc agcgtttgtg acccctgctc cgagttcgag
     1861 gacttttgga ggcccccttc tccctccgtg tctccagcgt cggagaagtc actgtgccgc
     1921 tctctggacg aagcccagcc ctacacgctg cctttcaagc cctatgcatg gagcggtctt
     1981 gctgggtctg acctgcggca cctggtgcag agctatcggc agtgcagcgc gctggagcgc
     2041 agcgcgggcc tgagcctctt ctgcgagcgc ggctcggagc cgggccgccc ggcagcgcgc
     2101 tacggccccg agcaggctgc gggcggagcc ggtgcgggac agccagggag ctgcggggtc
     2161 gccgggggcg ccaccagcgc tgcgggcctg gggctctacg gcgacttcgc gcctgcggcg
     2221 gccgggctgt acgagcggcc gagcacagca gcaggccggc tgtaccaaga tcatggccac
     2281 gagctgcacg cggacaagag cgtgggcgtc aaggtggagt cggagctgct ttgcacccgt
     2341 ctgctgctgg gcggcggctc ctacaaatgc atcaaatgca gcaaggtgtt ctccacaccg
     2401 cacgggctgg aggtgcacgt gcgccggtcc cacagcggca caagaccctt tgcgtgcgag
     2461 atgtgcggca agaccttcgg gcacgcggtg agcctggagc aacacaaggc agtgcactcc
     2521 caggaacgca gctttgactg taagatctgt ggcaagagct tcaagaggtc atccacgctg
     2581 tccacacatc tgctcattca ctcggacacc cggccctatc cctgtcagta ctgtggcaaa
     2641 agattccacc agaagtcaga tatgaagaaa cacaccttca tccacacagg tgagaagccc
     2701 cacaaatgcc aggtgtgcgg caaagccttc agtcagagct ccaacctcat cactcatagc
     2761 agaaagcaca caggcttcaa gccctttggc tgtgacctgt gtgggaaggg cttccagagg
     2821 aaggtggatc tcaggaggca ccgagagact cagcatggac tcaaatgact cgagcgggat
     2881 caattccgcc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     2941 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     3001 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     3061 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     3121 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     3181 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     3241 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     3301 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     3361 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     3421 gtggttttcc tttgaaaaac acgataatac catggtgagc aagggcgagg agctgttcac
     3481 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3541 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     3601 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     3661 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     3721 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3781 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3841 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3901 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     3961 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4021 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     4081 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4141 cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagag tcgacctgca
     4201 ggcatgcaag cttcaggtag ccggctaacg ttaacaaccg gtacctctag aactatagct
     4261 agcatgcgca aatttaaagc gctgatatcg ataaaataaa agattttatt tagtctccag
     4321 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     4381 ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa
     4441 cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     4501 ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa
     4561 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     4621 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     4681 gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc
     4741 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     4801 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc
     4861 ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa
     4921 ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc
     4981 gcagaaagag ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac
     5041 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5101 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5161 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5221 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5281 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5341 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5401 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5461 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5521 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5581 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5641 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5701 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5761 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5821 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5881 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5941 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6001 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6061 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6121 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6181 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6241 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6301 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6361 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6421 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6481 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6541 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6601 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6661 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6721 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6781 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6841 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6901 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6961 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7021 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7081 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7141 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     7201 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     7261 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     7321 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     7381 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     7441 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     7501 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     7561 cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga
     7621 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     7681 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     7741 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggc
//
