LOCUS       sequence-384396-        6809 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 6809)
  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 6809)
  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 E4orf1
FEATURES             Location/Qualifiers
     source          1..6809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    557..898
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             965..1381
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1311..1333
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1349..1365
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1394..1403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1403..1426
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1439..1465
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1481..1505
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1887..1911)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1924..2491
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1941..2493
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2085..2102)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2312..2331
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2505..3104
                     /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(2505..2524)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3001..3021
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3180..3694
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3856..3872
                     /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(3880..3886,3887..3904,3905..3910))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3925..3946
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4063..4080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4234..4822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4314..4333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4993..5784,5785..5853))
                     /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     5616..5635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5854..5958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6026..6044
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6082..6104)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6204..6223
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6417..6439
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6567..6586
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(6786..6809,1..493)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata
       61 gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat
      121 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
      181 cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
      241 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
      301 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
      361 cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct gtttgcatcc
      421 gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact gcccacctcg
      481 ggggtctttc atttggaggt tccaccgaga tttggagacc cctgcccagg gaccaccgac
      541 ccccccgccg ggaggtaagc tggccagcgg tcgtttcgtg tctgtctctg tctttgtgcg
      601 tgtttgtgcc ggcatctaat gtttgcgcct gcgtctgtac tagttagcta actagctctg
      661 tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag
      721 acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg
      781 gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc
      841 gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca
      901 gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa
      961 ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
     1021 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
     1081 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
     1141 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
     1201 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
     1261 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
     1321 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
     1381 cggaattaga tctgccacca tggactacaa ggatgacgat gacaagctcg atggaggata
     1441 cccatacgat gttccagatt acgctgctcg aggttatcaa acaagtttgt acaaaaaagt
     1501 tggcatggct gccgctgtgg aagcgctgta tgttgttctg gagcgggagg gtgctatttt
     1561 gcctaggcag gagggttttt caggtgttta tgtgtttttc tctcctatta attttgttat
     1621 acctcctatg ggggctgtaa tgttgtctct acgcctgcgg gtatgtattc ccccgggcta
     1681 tttcggtcgc tttttagcac tgaccgatgt gaatcaacct gatgtgttta ccgagtctta
     1741 cattatgact ccggacatga ccgaggagct gtcggtggtg ctttttaatc acggtgacca
     1801 gtttttttac ggtcacgccg gcatggccgt agtccgtctt atgcttataa gggttgtttt
     1861 tcctgttgta agacaggctt ctaatgtttc aactttcttg tacaaagtgg ttcgataacg
     1921 aattccgccc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     1981 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     2041 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     2101 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     2161 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     2221 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     2281 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     2341 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     2401 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     2461 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatgacc gagtacaagc
     2521 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     2581 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     2641 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     2701 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     2761 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     2821 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     2881 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     2941 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     3001 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     3061 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatgcatc gatgtcgacc
     3121 tgcagccaag cttatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     3181 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     3241 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
     3301 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     3361 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
     3421 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     3481 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     3541 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata
     3601 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     3661 agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga
     3721 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt
     3781 tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg
     3841 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3901 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3961 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4021 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4081 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4141 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4201 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4261 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4321 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4381 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4441 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4501 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4561 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4621 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4681 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4741 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4801 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4861 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4921 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4981 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5041 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5101 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5161 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5221 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5281 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5341 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5401 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5461 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5521 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5581 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5641 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5701 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5761 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5821 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5881 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5941 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6001 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6061 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6121 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     6181 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     6241 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     6301 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6361 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6421 gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac
     6481 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     6541 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     6601 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt
     6661 gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg
     6721 aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag
     6781 gggggaatga aagaccccac ctgtaggtt
//
