LOCUS       sequence-485923-        6971 bp    DNA     circular SYN 15-SEP-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  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 6971)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 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 BALF1
FEATURES             Location/Qualifiers
     source          1..6971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     regulatory      1418..1427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1427..1450
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1463..1489
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1505..1529
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2073..2097)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2110..2677
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2127..2679
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2271..2288)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2498..2517
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2691..3290
                     /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(2691..2710)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3187..3207
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3366..3880
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4042..4058
                     /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(4066..4072,4073..4090,4091..4096))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4111..4132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4249..4266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4420..5008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4500..4519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5179..5970,5971..6039))
                     /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     5802..5821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6040..6144)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6212..6230
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6268..6290)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6390..6409
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6603..6625
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6753..6772
                     /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 tagatctgcc accatggact acaaggatga
     1441 cgatgacaag ctcgatggag gatacccata cgatgttcca gattacgctg ctcgaggtta
     1501 tcaaacaagt ttgtacaaaa aagttggcat gaggccagcc aagtctacag attctgtgtt
     1561 tgtgaggacc ccggtcgagg cgtgggtcgc gccctcgccg ccggacgaca aggtggctga
     1621 gtccagctac ctcatgttca gggccatgta cgcggtgttc acccgggatg agaaagacct
     1681 gcctttgcca gccctggtcc tctgccggct catcaaggcc tccctgagga aggataggaa
     1741 gctgtacgcg gagctggcct gcaggacagc cgacatcggg ggcaaagaca cgcacgtacg
     1801 gctcatcatc agcgtcctgc gcgcagtgta caacgaccac tacgactact ggtcgcggct
     1861 cagggtggtg ctgtgctaca cagtggtgtt tgcggtgcga aactacctgg atgaccacaa
     1921 gagcgccgcc ttcgtgctgg gggcaatcgc ccactacctg gccctctatc gcagactctg
     1981 gtttgcgagg ctgggcggca tgccaagatc gctgagacgt cagttccccg tgacgtgggc
     2041 cctggccagc ctgactgact tcctgaaatc tttgccaact ttcttgtaca aagtggttcg
     2101 ataacgaatt ccgccccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     2161 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     2221 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     2281 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     2341 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     2401 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     2461 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     2521 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     2581 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     2641 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc atgaccgagt
     2701 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     2761 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     2821 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     2881 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     2941 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     3001 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     3061 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     3121 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     3181 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     3241 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga tgcatcgatg
     3301 tcgacctgca gccaagctta tcgataaaat aaaagatttt atttagtctc cagaaaaagg
     3361 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     3421 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     3481 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     3541 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     3601 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     3661 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     3721 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtgtat
     3781 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     3841 cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt
     3901 tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca
     3961 ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaaa
     4021 gagctgggga gaattgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4081 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5041 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5101 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5161 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5221 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5281 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5341 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5401 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5461 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5521 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5581 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5641 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5701 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5761 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5821 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5881 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5941 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6001 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6061 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6121 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6181 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6241 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6301 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6361 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6421 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     6481 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6541 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6601 ttcccagtca cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg
     6661 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     6721 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     6781 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc
     6841 caatttgtta aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac
     6901 cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag
     6961 aaaaaggggg g
//
