LOCUS Exported 7213 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of mouse Arf. ACCESSION . VERSION . KEYWORDS MSCV-mouse p19ARF-IRES-GFP 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7213 /organism="synthetic DNA construct" /mol_type="other DNA" 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" 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 2725..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 2725..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(4306..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(5419..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 7212..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 //