LOCUS       sequence-287989-        6947 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 6947)
  AUTHORS   Rozenblatt-Rosen O, Deo RC, Padi M, Adelmant G, Calderwood MA, 
            Rolland T, Grace M, Dricot A, Askenazi M, Tavares M, Pevzner SJ, 
            Abderazzaq F, Byrdsong D, Carvunis AR, Chen AA, Cheng J, Correll M, 
            Duarte M, Fan C, Feltkamp MC, Ficarro SB, Franchi R, Garg BK, 
            Gulbahce N, Hao T, Holthaus AM, James R, Korkhin A, Litovchick L, 
            Mar JC, Pak TR, Rabello S, Rubio R, Shen Y, Singh S, Spangle JM, 
            Tasan M, Wanamaker S, Webber JT, Roecklein-Canfield J, Johannsen E, 
            Barabasi AL, Beroukhim R, Kieff E, Cusick ME, Hill DE, Munger K, 
            Marto JA, Quackenbush J, Roth FP, DeCaprio JA, Vidal M
  TITLE     Interpreting cancer genomes using systematic host network 
            perturbations by tumour virus proteins.
  JOURNAL   Nature. 2012 Jul 26;487(7408):491-5.
  PUBMED    22810586
REFERENCE   2  (bases 1 to 6947)
  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: "Nature."; 
            date: "2012-07-26"; volume: "487(7408)"; pages: "491-5"
COMMENT     Sequence Label: MSCV-N SV40 ST
FEATURES             Location/Qualifiers
     source          1..6947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    568..909
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             976..1392
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1322..1344
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1360..1376
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1405..1414
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1414..1437
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1450..1476
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1492..1516
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1516..2037
                     /codon_start=1
                     /product="SV40 (simian virus 40) small t antigen"
                     /label=small t antigen
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEVFASSLNPGVDAMYCKQWPECA
                     KKMSANCICLLCLLRMKHENRKLYRKDPLVWVDCYCFDCFRMWFGLDLCEGTLLLWCDI
                     IGQTTYRDLKL"
     protein_bind    complement(2036..2060)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2073..2640
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2090..2642
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2234..2251)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2461..2480
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2654..3253
                     /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(2654..2673)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3150..3170
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3329..3843
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4005..4021
                     /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(4029..4035,4036..4053,4054..4059))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4074..4095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4212..4229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4383..4971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4463..4482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5142..5933,5934..6002))
                     /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     5765..5784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6003..6107)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6175..6193
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6231..6253)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6353..6372
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6566..6588
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6716..6735
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(6935..6947,1..504)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat
       61 aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc
      121 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
      181 ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
      241 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
      301 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg
      361 cgccagtcct ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc
      421 tgtttgcatc cgaatcgtgg actcgctgat ccttgggagg gtctcctcag attgattgac
      481 tgcccacctc gggggtcttt catttggagg ttccaccgag atttggagac ccctgcccag
      541 ggaccaccga cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct
      601 gtctttgtgc gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct
      661 aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc
      721 aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg
      781 gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta
      841 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct
      901 tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat
      961 ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg
     1021 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     1081 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     1141 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     1201 ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     1261 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     1321 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     1381 tctctaggcg ccggaattag atctgccacc atggactaca aggatgacga tgacaagctc
     1441 gatggaggat acccatacga tgttccagat tacgctgctc gaggttatca aacaagtttg
     1501 tacaaaaaag ttggcatgga taaagtttta aacagagagg aatctttgca gctaatggac
     1561 cttctaggtc ttgaaaggag tgcctggggg aatattcctc tgatgagaaa ggcatattta
     1621 aaaaaatgca aggagtttca tcctgataaa ggaggagatg aagaaaaaat gaagaaaatg
     1681 aatactctgt acaagaaaat ggaagatgga gtaaaatatg ctcatcaacc tgactttgga
     1741 ggcttctggg atgcaactga ggtatttgct tcttccttaa atcctggtgt tgatgcaatg
     1801 tactgcaaac aatggcctga gtgtgcaaag aaaatgtctg ctaactgcat atgcttgctg
     1861 tgcttactga ggatgaagca tgaaaataga aaattataca ggaaagatcc acttgtgtgg
     1921 gttgattgct actgcttcga ttgctttaga atgtggtttg gacttgatct ttgtgaagga
     1981 accttacttc tgtggtgtga cataattgga caaactacct acagagattt aaagctctca
     2041 actttcttgt acaaagtggt tcgataacga attccgcccc ccccccctaa cgttactggc
     2101 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     2161 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     2221 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     2281 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     2341 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     2401 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     2461 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     2521 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     2581 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     2641 tatggccaca accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     2701 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     2761 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     2821 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     2881 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     2941 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     3001 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     3061 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     3121 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     3181 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     3241 gcccggtgcc tgatgcatcg atgtcgacct gcagccaagc ttatcgataa aataaaagat
     3301 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     3361 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     3421 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     3481 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     3541 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     3601 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     3661 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     3721 gcgtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     3781 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     3841 catgggtaac agtttcttga agttggagaa caacattctg agggtaggag tcgaatatta
     3901 agtaatcctg actcaattag ccactgtttt gaatccacat actccaatac tcctgaaata
     3961 gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta tccgctcaca
     4021 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4081 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4141 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4201 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4261 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4321 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4381 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4441 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4501 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4561 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4621 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4681 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4741 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4801 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4861 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4921 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4981 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5041 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5101 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5161 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5221 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5281 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5341 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5401 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5461 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5521 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5581 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5641 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5701 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5761 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5821 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5881 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5941 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6001 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6061 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6121 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6181 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6241 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6301 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     6361 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     6421 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6481 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6541 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga
     6601 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     6661 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     6721 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     6781 gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc
     6841 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat
     6901 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agacccc
//
