LOCUS       sequence-466643-        7088 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 7088)
  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 7088)
  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 BARF1
FEATURES             Location/Qualifiers
     source          1..7088
                     /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(2190..2214)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2227..2794
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2244..2796
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2388..2405)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2615..2634
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2808..3407
                     /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(2808..2827)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3304..3324
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3483..3997
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4159..4175
                     /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(4183..4189,4190..4207,4208..4213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4228..4249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4366..4383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4537..5125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4617..4636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5296..6087,6088..6156))
                     /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     5919..5938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6157..6261)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6329..6347
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6385..6407)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6507..6526
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6720..6742
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6870..6889
                     /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 ggccaggttc atcgctcagc tcctcctgtt
     1561 ggcctcctgt gtggccgccg gccaggctgt caccgctttc ttgggtgagc gagtcaccct
     1621 gacctcctac tggaggaggg tgagcctcgg tccagagatt gaggtcagct ggtttaaact
     1681 gggcccagga gaggagcagg tgcttattgg gcgcatgcac cacgatgtca tctttataga
     1741 gtggcctttc aggggcttct ttgatatcca cagaagtgcc aacaccttct ttttagtagt
     1801 caccgctgcc aacatctccc atgacggcaa ctacctgtgc cgcatgaaac tgggcgagac
     1861 cgaggtcacc aagcaggaac acctgagcgt ggtgaagcct ctaacgctgt ctgtccactc
     1921 cgaaaggtct cagttcccag acttctctgt ccttactgtg acatgcaccg tgaatgcatt
     1981 tccccatccc cacgtccagt ggctcatgcc cgagggcgtg gagcccgcac caactgcggc
     2041 aaatggcggt gttatgaagg aaaaggatgg gagcctctct gttgctgttg acctgtcact
     2101 tcccaagccc tggcacctgc cagtgacctg cgttgggaaa aatgacaagg aggaagccca
     2161 cggggtttat gtttctggat acttgtcgca accaactttc ttgtacaaag tggttcgata
     2221 acgaattccg cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     2281 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     2341 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     2401 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     2461 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     2521 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     2581 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     2641 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     2701 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     2761 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg accgagtaca
     2821 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     2881 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     2941 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     3001 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     3061 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     3121 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     3181 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     3241 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     3301 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     3361 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatgc atcgatgtcg
     3421 acctgcagcc aagcttatcg ataaaataaa agattttatt tagtctccag aaaaaggggg
     3481 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     3541 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     3601 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3661 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     3721 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3781 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3841 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca
     3901 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     3961 ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg
     4021 agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg
     4081 ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc gcagaaagag
     4141 ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6301 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     6361 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6421 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6481 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6541 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6601 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6661 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6721 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
     6781 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     6841 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     6901 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
     6961 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
     7021 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
     7081 aagggggg
//
