LOCUS Exported 9036 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION retroviral expression of Cre recombinase. ACCESSION . VERSION . KEYWORDS MSCV-PIG-Cre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9036) AUTHORS Chen Z, Trotman LC, Shaffer D, Lin HK, Dotan ZA, Niki M, Koutcher JA, Scher HI, Ludwig T, Gerald W, Cordon-Cardo C, Pandolfi PP TITLE Crucial role of p53-dependent cellular senescence in suppression of Pten-deficient tumorigenesis. JOURNAL Nature. 2005 Aug 4;436(7051):725-30. PUBMED 16079851 REFERENCE 2 (bases 1 to 9036) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9036 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..517 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 581..922 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 989..1405 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1335..1357 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1373..1389 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1426..1446 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1447..2475 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED GD" promoter 2849..3348 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(2884..2907) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 3281..3300 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 3369..3968 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3369..3388) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3865..3885 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 4046..4598 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4190..4207) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4417..4436 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4610..5329 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4655..4676) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4916..4935) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5263..5284 /label=EGFP-C /note="EGFP, forward primer" LTR 5431..5945 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 6107..6123 /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(6131..6161) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6176..6197 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6314..6331) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6485..7073) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6565..6584) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7244..8104) /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 7867..7886 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8105..8209) /gene="bla" /label=AmpR promoter primer_bind 8277..8295 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(8333..8355) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 8455..8474 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8668..8690 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8818..8837 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc 61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag 121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa 181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt 241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt 301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac 361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa 421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct 481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga 541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt 601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct 661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg 721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc 781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg 841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg 901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac 961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac 1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa 1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg 1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg 1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga 1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc 1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc 1381 agccctcact ccttctctag gcgccggaat tagatctgga ccatgcccaa gaagaagagg 1441 aaggtgtcca atttactgac cgtacaccaa aatttgcctg cattaccggt cgatgcaacg 1501 agtgatgagg ttcgcaagaa cctgatggac atgttcaggg atcgccaggc gttttctgag 1561 catacctgga aaatgcttct gtccgtttgc cggtcgtggg cggcatggtg caagttgaat 1621 aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg attatcttct atatcttcag 1681 gcgcgcggtc tggcagtaaa aactatccag caacatttgg gccagctaaa catgcttcat 1741 cgtcggtccg ggctgccacg accaagtgac agcaatgctg tttcactggt tatgcggcgg 1801 atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc 1861 actgatttcg accaggttcg ttcactcatg gaaaatagcg atcgctgcca ggatatacgt 1921 aatctggcat ttctggggat tgcttataac accctgttac gtatagccga aattgccagg 1981 atcagggtta aagatatctc acgtactgac ggtgggagaa tgttaatcca tattggcaga 2041 acgaaaacgc tggttagcac cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa 2101 ctggtcgagc gatggatttc cgtctctggt gtagctgatg atccgaataa ctacctgttt 2161 tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct atcaactcgc 2221 gccctggaag ggatttttga agcaactcat cgattgattt acggcgctaa ggatgactct 2281 ggtcagagat acctggcctg gtctggacac agtgcccgtg tcggagccgc gcgagatatg 2341 gcccgcgctg gagtttcaat accggagatc atgcaagctg gtggctggac caatgtaaat 2401 attgtcatga actatatccg taacctggat agtgaaacag gggcaatggt gcgcctgctg 2461 gaagatggcg attagccatt aacgcgtaaa tgattgctat aattatttga tatttatggt 2521 gacatatgag aaaggatttc aacatcgacg gaaaatatgt agtgctgtct gtaagcacta 2581 atattcagtc gccagccgtc attgtcactg taaagctgag cgatagaatg cctgatattg 2641 actcaatatc cgttgcgttt cctgtcaaaa gtatgcgtag tgctgaacat ttcgtgatga 2701 atgccaccga ggaagaagca cggcgcggtt ttgctaaagt gatgtctgag tttggcgaac 2761 tcttgggtaa ggttggaatt gtcgaggctg ggtgtggcgg acggtaccga gctcgaattc 2821 agatctctcg aggttaacga attctaccgg gtaggggagg cgcttttccc aaggcagtct 2881 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct 2941 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc 3001 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg 3061 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc 3121 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc 3181 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc 3241 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag 3301 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca 3361 agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca 3421 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg 3481 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg 3541 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca 3601 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt 3661 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc 3721 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg 3781 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg 3841 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg 3901 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg 3961 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg 4021 catcgaattc cgcccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg 4081 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc 4141 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa 4201 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga 4261 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc 4321 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc 4381 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac 4441 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg 4501 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac 4561 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa 4621 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa 4681 cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat gccacctacg gcaagctgac 4741 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac 4801 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt 4861 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga 4921 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat 4981 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta 5041 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggc 5101 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca 5161 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac 5221 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt 5281 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat gaattaatta 5341 agaattatca agcttatcga taccgtcgac ctgcagccaa gcttatcgat aaaataaaag 5401 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag 5461 ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga atagagaagt 5521 tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga tatctgtggt 5581 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc 5641 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc 5701 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc 5761 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgataga 5821 ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg 5881 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct 5941 ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat 6001 taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa 6061 tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt tatccgctca 6121 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 6181 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 6241 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 6301 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 6361 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 6421 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 6481 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 6541 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 6601 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 6661 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 6721 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 6781 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 6841 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 6901 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 6961 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 7021 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 7081 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 7141 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 7201 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 7261 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 7321 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 7381 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 7441 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 7501 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 7561 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 7621 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 7681 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 7741 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 7801 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 7861 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 7921 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 7981 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 8041 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8101 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 8161 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 8221 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata 8281 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 8341 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 8401 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 8461 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 8521 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 8581 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 8641 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggcgcaag 8701 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat 8761 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt 8821 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat 8881 gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag tggtccaggc 8941 tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc catagataaa 9001 ataaaagatt ttatttagtc tccagaaaaa gggggg //