LOCUS       sequence-248369-        7916 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 7916)
  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 7916)
  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 LMP2A
FEATURES             Location/Qualifiers
     source          1..7916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..72
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             272..788
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    852..1193
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1260..1676
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1606..1628
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1644..1660
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1689..1698
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1698..1721
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1734..1760
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1776..1800
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3289..3313)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3326..3893
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3343..3895
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3487..3504)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3714..3733
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3907..4506
                     /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(3907..3926)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4403..4423
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4582..5096
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5258..5274
                     /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(5282..5288,5289..5306,5307..5312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5327..5348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5465..5482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5636..6224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5716..5735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6395..7186,7187..7255))
                     /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     7018..7037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7256..7360)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7428..7446
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7484..7506)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7606..7625
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7819..7841
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
       61 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
      121 cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag
      181 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
      241 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
      301 agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa
      361 gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg
      421 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc
      481 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
      541 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
      601 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata
      661 gactgcgtcg cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgaatc
      721 gtggactcgc tgatccttgg gagggtctcc tcagattgat tgactgccca cctcgggggt
      781 ctttcatttg gaggttccac cgagatttgg agacccctgc ccagggacca ccgacccccc
      841 cgccgggagg taagctggcc agcggtcgtt tcgtgtctgt ctctgtcttt gtgcgtgttt
      901 gtgccggcat ctaatgtttg cgcctgcgtc tgtactagtt agctaactag ctctgtatct
      961 ggcggacccg tggtggaact gacgagttct gaacacccgg ccgcaaccct gggagacgtc
     1021 ccagggactt tgggggccgt ttttgtggcc cgacctgagg aagggagtcg atgtggaatc
     1081 cgaccccgtc aggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc
     1141 cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct
     1201 gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg
     1261 gccagactgt taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga
     1321 tcgctcacaa ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag
     1381 aatggccaac ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca
     1441 cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct
     1501 acatcgtgac ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac
     1561 accctaagcc tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc
     1621 gttcgacccc gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccggaa
     1681 ttagatctgc caccatggac tacaaggatg acgatgacaa gctcgatgga ggatacccat
     1741 acgatgttcc agattacgct gctcgaggtt atcaaacaag tttgtacaaa aaagttggca
     1801 tggggtccct agaaatggtg ccaatgggcg cgggtccccc tagccccggc ggggatccgg
     1861 atgggtacga tggcggaaac aactcccaat atccatctgc ttctggctct tctgggaaca
     1921 cccccacccc accgaacgat gaggaacgtg aatctaatga agagccccca ccgccttatg
     1981 aggacccata ttggggcaat ggcgaccgtc actcggacta tcaaccacta ggaacccaag
     2041 atcaaagtct gtacttggga ttgcaacacg acgggaatga cgggctccct ccccctccct
     2101 actctccacg ggatgactca tctcaacaca tatacgaaga agcgggcaga ggaagtatga
     2161 atccagtatg cctgcctgta attgttgcgc cctacctctt ttggctggcg gctattgccg
     2221 cctcgtgttt cacggcctca gttagtaccg ttgtgaccgc caccggcttg gccctctcac
     2281 ttctactctt ggcagcagtg gccagctcat atgccgctgc acaaaggaaa ctgctgacac
     2341 cggtgacagt gcttactgcg gttgtcactt tctttgcaat ttgcctaaca tggaggattg
     2401 aggacccacc ttttaattct cttctgtttg cattgctggc cgcagctggc ggactacaag
     2461 gcatttacgt tctggtgatg cttgtgctcc tgatactagc gtacagaagg agatggcgcc
     2521 gtttgactgt ttgtggcggc atcatgtttt tggcatgtgt acttgtcctc atcgtcgacg
     2581 ctgttttgca gctgagtccc ctccttggag ctgtaactgt ggtttccatg acgctgctgc
     2641 tactggcttt cgtcctctgg ctctcttcgc cagggggcct aggtactctt ggtgcagccc
     2701 ttttaacatt ggcagcagct ctggcactgc tagcgtcact gattttgggc acacttaact
     2761 tgactacaat gttccttctc atgctcctat ggacacttgt ggttctcctg atttgctctt
     2821 cgtgctcttc atgtccactg agcaagatcc ttctggcacg actgttccta tatgctctcg
     2881 cactcttgtt gctagcctcc gcgctaatcg ctggtggcag tattttgcaa acaaacttca
     2941 agagtttaag cagcactgaa tttataccca atttgttctg catgttatta ctgattgtcg
     3001 ctggcatact cttcattctt gctatcctga ccgaatgggg cagtggaaat agaacatacg
     3061 gtccagtttt tatgtgcctc ggtggcctgc tcaccatggt agccggcgct gtgtggctga
     3121 cggtgatgtc taacacgctt ttgtctgcct ggattcttac agcaggattc ctgattttcc
     3181 tcattggctt tgccctcttt ggggtcatta gatgctgccg ctactgctgc tactactgcc
     3241 ttacactgga aagtgaggag cgcccaccga ccccatatcg caacactgta ccaactttct
     3301 tgtacaaagt ggttcgataa cgaattccgc cccccccccc taacgttact ggccgaagcc
     3361 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     3421 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     3481 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     3541 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     3601 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     3661 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     3721 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     3781 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     3841 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     3901 acaaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     3961 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     4021 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     4081 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     4141 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4201 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     4261 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4321 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     4381 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4441 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4501 gcctgatgca tcgatgtcga cctgcagcca agcttatcga taaaataaaa gattttattt
     4561 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     4621 gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa
     4681 ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc
     4741 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt
     4801 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     4861 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     4921 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc
     4981 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     5041 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatgggt
     5101 aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata ttaagtaatc
     5161 ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa atagttcatt
     5221 atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc acaattccac
     5281 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5341 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5401 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5461 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5521 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5581 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5641 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5701 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5761 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5821 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5881 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5941 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6001 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6061 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6121 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6181 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6241 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6301 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6361 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6421 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6481 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6541 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6601 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6661 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6721 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6781 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6841 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6901 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6961 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7021 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7081 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7141 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7201 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7261 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7321 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7381 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     7441 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     7501 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     7561 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     7621 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     7681 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     7741 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     7801 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc
     7861 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca taccca
//
