LOCUS       sequence-8797-ex        7233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7233)
  AUTHORS   Swarbrick A, Roy E, Allen T, Bishop JM
  TITLE     Id1 cooperates with oncogenic Ras to induce metastatic mammary 
            carcinoma by subversion of the cellular senescence response.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Apr 8. 105(14):5402-7.
  PUBMED    18378907
REFERENCE   2  (bases 1 to 7233)
  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: "Proc Natl 
            Acad Sci U S A. 2008 Apr 8. 105(14):5402-7."
COMMENT     Sequence Label: pRQ-hRasV12
FEATURES             Location/Qualifiers
     source          1..7233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..530
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(66..89)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     463..482
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             551..1150
                     /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(551..570)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1047..1067
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1331..1756
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(1859..1881)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2017..2037)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2022..2351
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2202..2337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2373..2392
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2473..2490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2644..3232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2724..2743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3403..4194,4195..4263))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4026..4045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4264..4368)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4436..4454
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        4554..4934
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4935..5138
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5139..5314
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5377..5576
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5777..6193
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6123..6145
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6161..6177
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        6224..6538
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    6232..6250
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6268..6286
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6303..6321
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6339..6357
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6375..6393
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6410..6428
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6446..6464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6511..6535
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6549..6573
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6619..7188
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=H-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     protein_bind    complement(join(7223..7233,1..14))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggtgatgtg gaattctacc gggtagggga ggcgcttttc ccaaggcagt
       61 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
      121 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
      181 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
      241 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
      301 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
      361 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
      421 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
      481 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
      541 caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
      601 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
      661 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
      721 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
      781 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
      841 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
      901 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
      961 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     1021 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     1081 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     1141 cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca
     1201 tgcatcggcg tctcgagata tcagtggtcc aggctctagt tttgactcaa caatatcacc
     1261 agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt agtctccaga
     1321 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     1381 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     1441 cagatggaac agggtcgacc ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     1501 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     1561 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc
     1621 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     1681 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     1741 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
     1801 gacccaccac cgggaggtaa gctggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     1861 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     1921 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     1981 agtcacgtag cgatagcgga gtgtagatcc ggctgtggaa tgtgtgtcag ttagggtgtg
     2041 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2101 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2161 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2221 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2281 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2341 gcttttgcaa aaagcttact ggcttaacta tgcggcatca gagcagattg tactgagagt
     2401 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     2461 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2521 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2581 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2641 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2701 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2761 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2821 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2881 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2941 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3001 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3061 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     3121 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3181 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3241 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3301 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3361 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3421 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3481 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3541 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3601 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3661 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
     3721 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3781 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3841 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3901 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3961 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     4021 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4081 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4141 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4201 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4261 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4321 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4381 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     4441 gcgtatcacg aggccctttc gtcttcaaga attagcttgg ccattgcata cgttgtatcc
     4501 atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg
     4561 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     4621 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     4681 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     4741 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     4801 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg ctctggcatt
     4861 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4921 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4981 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     5041 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     5101 gtaggcgtgt acggtgggag gtctatataa gcagagctca ataaaagagc ccacaacccc
     5161 tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta ttcccaataa
     5221 agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg gtctcctctg
     5281 agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg atttggagac
     5341 ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa cttatctgtg
     5401 tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt actagttagc
     5461 taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa cacccggccg
     5521 caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga cctgaggaag
     5581 ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag acgagaacct
     5641 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc
     5701 ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta
     5761 tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct taggtcactg
     5821 gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga gacgttgggt
     5881 taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag acggcacctt
     5941 taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc cgcatggaca
     6001 cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc cccctccctg
     6061 ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg ccccgtctct
     6121 cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag ccctcactcc
     6181 ttctctaggc gccggaattg aagatctcct ttcgtcttca atcgagttta ctccctatca
     6241 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
     6301 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
     6361 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
     6421 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag
     6481 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     6541 gacacatcac aagtttgtac aaaaaagcag gctctttaaa ggaaccaatt cagtcgactg
     6601 gatccggtac cgaattcgat gacggaatat aagctggtgg tggtgggcgc cggcggtgtg
     6661 ggcaagagtg cgctgaccat ccagctgatc cagaaccatt ttgtggacga atacgacccc
     6721 actatagagg attcctaccg gaagcaggtg gtcattgatg gggagacgtg cctgttggac
     6781 atcctggata ccgccggcca ggaggagtac agcgccatgc gggaccagta catgcgcacc
     6841 ggggagggct tcctgtgtgt gtttgccatc aacaacacca agtcttttga ggacatccac
     6901 cagtacaggg agcagatcaa acgggtgaag gactcggatg acgtgcccat ggtgctggtg
     6961 gggaacaagt gtgacctggc tgcacgcact gtggaatctc ggcaggctca ggacctcgcc
     7021 cgaagctacg gcatccccta catcgagacc tcggccaaga cccggcaggg agtggaggat
     7081 gccttctaca cgttggtgcg tgagatccgg cagcacaagc tgcggaagct gaaccctcct
     7141 gatgagagtg gccccggctg catgagctgc aagtgtgtgc tctcctgaga attcaattcg
     7201 cggccgcact cgagatatct agacccagct ttc
//
