LOCUS       sequence-207628-        7305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse Arf.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7305)
  AUTHORS   Kuo ML, Duncavage EJ, Mathew R, den Besten W, Pei D, Naeve D, 
            Yamamoto T, Cheng C, Sherr CJ, Roussel MF
  TITLE     Arf induces p53-dependent and -independent antiproliferative genes.
  JOURNAL   Cancer Res. 2003 Mar 1;63(5):1046-53.
  PUBMED    12615721
REFERENCE   2  (bases 1 to 7305)
  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: "Cancer 
            Res."; date: "2003-03-1"; volume: "63(5)"; pages: "1046-53"
COMMENT     Sequence Label: MSCV-mouse p19ARF-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..136
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             336..852
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    916..1257
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1324..1740
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1670..1692
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1708..1724
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2602..3151
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2746..2763)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2973..2992
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3154..3156,3157..3159,3160..3873)
                     /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(3154..3156,3157..3159,3160..3870)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3199..3220)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3460..3479)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3807..3828
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4035..4549
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4711..4727
                     /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(4735..4741,4742..4759,4760..4765))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4780..4801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4918..4935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5089..5677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5169..5188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5848..6639,6640..6708))
                     /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     6471..6490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6709..6813)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6881..6899
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6937..6959)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7059..7078
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7272..7294
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
       61 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
      121 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
      181 cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct
      241 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
      301 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
      361 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag
      421 agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc
      481 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
      541 ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
      601 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
      661 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
      721 gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg
      781 aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg
      841 gggtctttca tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc
      901 cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt
      961 gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt
     1021 atctggcgga cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga
     1081 cgtcccaggg actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg
     1141 aatccgaccc cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg
     1201 cctccgtctg aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag
     1261 cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat
     1321 tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
     1381 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
     1441 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
     1501 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
     1561 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
     1621 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
     1681 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
     1741 ggaattccgg gaatttcttc actttgacat tcagagcact gggcagaaat cacatcgcct
     1801 caacgccgaa ggggctgggg gcggcgcttc tcacctcgct tgtcacagtg aggccgccgc
     1861 tgagggagta cagcagcggg agcatgggtc gcaggttctt ggtcactgtg aggattcagc
     1921 gcgcgggccg cccactccaa gagagggttt tcttggtgaa gttcgtgcga tcccggagac
     1981 ccaggacagc gagctgcgct ctggctttcg tgaacatgtt gttgaggcta gagaggatct
     2041 tgagaagagg gccgcaccgg aatcctggac caggtgatga tgatgggcaa cgttcacgta
     2101 gcagctcttc tgctcaacta cggtgcagat tcgaactgcg aggaccccac taccttctcc
     2161 cgcccggtgc acgacgcagc gcgggaaggc ttcctggaca cgctggtggt gctgcacggg
     2221 tcaggggctc ggctggatgt gcgcgatgcc tggggtcgcc tgccgctcga cttggcccaa
     2281 gagcggggac atcaagacat cgtgcgatat ttgcgttccg ctgggtgctc tttgtgttcc
     2341 gctgggtggt ctttgtgtac cgctgggaac gtcgcccaga ccgacgggca tagcttcagc
     2401 tcaagcacgc ccagggccct ggaacttcgc ggccaatccc aagagcagag ctaaatccgg
     2461 cctcagcccg cctttttctt cttagcttca cttctagcga tgctagcgtg tctagcatgt
     2521 ggctttaaaa aatacataat aatgcttttt ttttgccgga attcgttaac ctcgagcggg
     2581 atcaattccg ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     2641 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     2701 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     2761 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     2821 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     2881 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     2941 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     3001 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     3061 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     3121 acgtggtttt cctttgaaaa acacgataat accatggtga gcaagggcga ggagctgttc
     3181 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     3241 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     3301 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     3361 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3421 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     3481 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3541 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3601 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     3661 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3721 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     3781 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3841 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag agtcgacctg
     3901 caggcatgca agcttcaggt agccggctaa cgttaacaac cggtacctct agaactatag
     3961 ctagcatgcg caaatttaaa gcgctgatat cgataaaata aaagatttta tttagtctcc
     4021 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     4081 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
     4141 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     4201 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
     4261 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     4321 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     4381 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
     4441 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     4501 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt
     4561 tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta atcctgactc
     4621 aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc attatggaca
     4681 gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc cacacaacat
     4741 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4801 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4861 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4921 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4981 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5041 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5101 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5161 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5221 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5281 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5341 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5401 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5461 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5521 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5581 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5641 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5701 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5761 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5821 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5881 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5941 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6001 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6061 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6121 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6181 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6241 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6301 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6361 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6421 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6481 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6541 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6601 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6661 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6721 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6781 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6841 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6901 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6961 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     7021 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     7081 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     7141 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     7201 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     7261 gccagggttt tcccagtcac gacgttgtaa aacgacggcg caagg
//
