LOCUS       sequence-203896-        7213 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 7213)
  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 7213)
  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..7213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2168..2716
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2312..2329)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2539..2558
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2725..2727,2728..2730,2731..3444)
                     /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(2725..2727,2728..2730,2731..3441)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2770..2791)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3031..3050)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3378..3399
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3606..4120
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4282..4298
                     /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(4306..4312,4313..4330,4331..4336))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4351..4372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4489..4506)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4660..5248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4740..4759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5419..6210,6211..6279))
                     /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     6042..6061
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6280..6384)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6452..6470
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6508..6530)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6630..6649
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6843..6865
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6993..7012
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7212..7213,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaattc gtcacagtga ggccgccgct gagggagtac
     1441 agcagcggga gcatgggtcg caggttcttg gtcactgtga ggattcagcg cgcgggccgc
     1501 ccactccaag agagggtttt cttggtgaag ttcgtgcgat cccggagacc caggacagcg
     1561 agctgcgctc tggctttcgt gaacatgttg ttgaggctag agaggatctt gagaagaggg
     1621 ccgcaccgga atcctggacc aggtgatgat gatgggcaac gttcacgtag cagctcttct
     1681 gctcaactac ggtgcagatt cgaactgcga ggaccccact accttctccc gcccggtgca
     1741 cgacgcagcg cgggaaggct tcctggacac gctggtggtg ctgcacgggt caggggctcg
     1801 gctggatgtg cgcgatgcct ggggtcgcct gccgctcgac ttggcccaag agcggggaca
     1861 tcaagacatc gtgcgatatt tgcgttccgc tgggtgctct ttgtgttccg ctgggtggtc
     1921 tttgtgtacc gctgggaacg tcgcccagac cgacgggcat agcttcagct caagcacgcc
     1981 cagggccctg gaacttcgcg gccaatccca agagcagagc taaatccggc ctcagcccgc
     2041 ctttttcttc ttagcttcac ttctagcgat gctagcgtgt ctagcatgtg gctttaaaaa
     2101 atacataata atgctttttt tttgaattct cacctcacgt gacgggatca attccgcccc
     2161 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     2221 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     2281 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     2341 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     2401 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     2461 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     2521 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     2581 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     2641 gtttagtcga ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     2701 gaaaaacacg ataatacctg tacaatggtg agcaagggcg aggagctgtt caccggggtg
     2761 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2821 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2881 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2941 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3001 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3061 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3121 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3181 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3241 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3301 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3361 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3421 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gagtcgacct gcaggcatgc
     3481 aagcttcagg tagccggcta acgttaacaa ccggtacctc tagaactata gctagcatgc
     3541 gcaaatttaa agcgctgata tcgataaaat aaaagatttt atttagtctc cagaaaaagg
     3601 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     3661 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     3721 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     3781 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     3841 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     3901 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     3961 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat
     4021 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     4081 cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt
     4141 tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca
     4201 ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaaa
     4261 gagctgggga gaattgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4321 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4381 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4441 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4501 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4561 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4621 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4681 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4741 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4801 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4861 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4921 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4981 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5041 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5101 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5161 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5221 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5281 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5341 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5401 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5461 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5521 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5581 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5641 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5701 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5761 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5821 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5881 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5941 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6001 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6061 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6121 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6181 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6241 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6301 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6361 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6421 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6481 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6541 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6601 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6661 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     6721 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6781 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6841 ttcccagtca cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg
     6901 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     6961 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     7021 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc
     7081 caatttgtta aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac
     7141 cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag
     7201 aaaaaggggg gaa
//
