LOCUS       sequence-270748-        9465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of multiple human genes (BCL2 and BCL6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9465)
  AUTHORS   Caeser R, Di Re M, Krupka JA, Gao J, Lara-Chica M, Dias JML, Cooke 
            SL, Fenner R, Usheva Z, Runge HFP, Beer PA, Eldaly H, Pak HK, Park 
            CS, Vassiliou GS, Huntly BJP, Mupo A, Bashford-Rogers RJM, Hodson DJ
  TITLE     Genetic modification of primary human B cells to model high-grade 
            lymphoma.
  JOURNAL   Nat Commun. 2019 Oct 4;10(1):4543. doi: 10.1038/s41467-019-12494-x.
  PUBMED    31586074
REFERENCE   2  (bases 1 to 9465)
  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: "Nat 
            Commun."; date: "2019-10-4"; volume: "10(1):4543. doi"; pages: " 
            10.1038/s41467-019-12494-x"
COMMENT     Sequence Label: MSCV-BCL6-t2A-BCL2
FEATURES             Location/Qualifiers
     source          1..9465
                     /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      1427..1436
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3572..3625
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    4382..4949
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4399..4951
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4543..4560)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4770..4789
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             5860..6374
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6536..6552
                     /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(6560..6566,6567..6584,6585..6590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6605..6626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6743..6760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6914..7502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6994..7013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7673..8464,8465..8533))
                     /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     8296..8315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8534..8638)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8706..8724
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8762..8784)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8884..8903
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9097..9119
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9247..9266
                     /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 tagatctctc gaggttgcca ccatggcctc
     1441 gccggctgac agctgtatcc agttcacccg ccatgccagt gatgttcttc tcaaccttaa
     1501 tcgtctccgg agtcgagaca tcttgactga tgttgtcatt gttgtgagcc gtgagcagtt
     1561 tagagcccat aaaacggtcc tcatggcctg cagtggcctg ttctatagca tctttacaga
     1621 ccagttgaaa tgcaacctta gtgtgatcaa tctagatcct gagatcaacc ctgagggatt
     1681 ctgcatcctc ctggacttca tgtacacatc tcggctcaat ttgcgggagg gcaacatcat
     1741 ggctgtgatg gccacggcta tgtacctgca gatggagcat gttgtggaca cttgccggaa
     1801 gtttattaag gccagtgaag cagagatggt ttctgccatc aagcctcctc gtgaagagtt
     1861 cctcaacagc cggatgctga tgccccaaga catcatggcc tatcggggtc gtgaggtggt
     1921 ggagaacaac ctgccactga ggagcgcccc tgggtgtgag agcagagcct ttgcccccag
     1981 cctgtacagt ggcctgtcca caccgccagc ctcttattcc atgtacagcc acctccctgt
     2041 cagcagcctc ctcttctccg atgaggagtt tcgggatgtc cggatgcctg tggccaaccc
     2101 cttccccaag gagcgggcac tcccatgtga tagtgccagg ccagtccctg gtgagtacag
     2161 ccggccgact ttggaggtgt cccccaatgt gtgccacagc aatatctatt cacccaagga
     2221 aacaatccca gaagaggcac gaagtgatat gcactacagt gtggctgagg gcctcaaacc
     2281 tgctgccccc tcagcccgaa atgcccccta cttcccttgt gacaaggcca gcaaagaaga
     2341 agagagaccc tcctcggaag atgagattgc cctgcatttc gagcccccca atgcacccct
     2401 gaaccggaag ggtctggtta gtccacagag cccccagaaa tctgactgcc agcccaactc
     2461 gcccacagag tcctgcagca gtaagaatgc ctgcatcctc caggcttctg gctcccctcc
     2521 agccaagagc cccactgacc ccaaagcctg caactggaag aaatacaagt tcatcgtgct
     2581 caacagcctc aatcagaatg ccaaaccaga ggggcctgag caggctgagc tgggccgcct
     2641 ttccccacga gcctacacgg ccccacctgc ctgccagcca cccatggagc ctgagaacct
     2701 tgacctccag tccccaacca agctgagtgc cagcggggag gactccacca tcccacaagc
     2761 cagccggctc aataacatcg ttaacaggtc catgacgggc tctccccgca gcagcagcga
     2821 gagccactca ccactctaca tgcacccccc gaagtgcacg tcctgcggct ctcagtcccc
     2881 acagcatgca gagatgtgcc tccacaccgc tggccccacg ttccctgagg agatgggaga
     2941 gacccagtct gagtactcag attctagctg tgagaacggg gccttcttct gcaatgagtg
     3001 tgactgccgc ttctctgagg aggcctcact caagaggcac acgctgcaga cccacagtga
     3061 caaaccctac aagtgtgacc gctgccaggc ctccttccgc tacaagggca acctcgccag
     3121 ccacaagacc gtccataccg gtgagaaacc ctatcgttgc aacatctgtg gggcccagtt
     3181 caaccggcca gccaacctga aaacccacac tcgaattcac tctggagaga agccctacaa
     3241 atgcgaaacc tgcggagcca gatttgtaca ggtggcccac ctccgtgccc atgtgcttat
     3301 ccacactggt gagaagccct atccctgtga aatctgtggc acccgtttcc ggcaccttca
     3361 gactctgaag agccacctgc gaatccacac aggagagaaa ccttaccatt gtgagaagtg
     3421 taacctgcat ttccgtcaca aaagccagct gcgacttcac ttgcgccaga agcatggcgc
     3481 catcaccaac accaaggtgc aataccgcgt gtcagccact gacctgcctc cggagctccc
     3541 caaagcctgc aacggatccg gcagcggcgg ggaaggaagg ggctccctgc tcacctgcgg
     3601 cgacgtggaa gagaaccccg gacctatggc gcacgctggg agaacagggt acgataaccg
     3661 ggagatagtg atgaagtaca tccattataa gctgtcgcag aggggctacg agtgggatgc
     3721 gggagatgtg ggagccgctc ctcctggtgc cgctcccgca ccgggcatct tctcctccca
     3781 gcctgggcac acgcctcatc cagccgcatc ccgggatcct gtcgccagga cctcgcctct
     3841 gcagacaccg gctgctcctg gagctgctgc aggacctgcg ctcagcccgg tgccacctgt
     3901 ggtccacctg accctccgcc aggccggcga cgacttctcc cgccgctacc gccgcgactt
     3961 cgccgagatg tccagccagc tgcacctgac gcccttcacc gcgcggggac gctttgccac
     4021 ggtggtggag gagctcttca gggacggggt gaactggggg aggattgtgg ccttctttga
     4081 gttcggtggg gtcatgtgtg tggagagcgt caaccgggag atgtcgcccc tggtggacaa
     4141 catcgccctg tggatgactg agtacctgaa ccggcacctg cacacctgga tccaggataa
     4201 cggaggctgg gatgcctttg tggaactgta cggccccagc atgcggcctc tgtttgattt
     4261 ctcctggctg tctctgaaga ctctgctcag tttggccctg gtgggagctt gcatcaccct
     4321 gggtgcctat ctgggccaca agtgatagaa cgcggccgca ccggtggatc ctacgtagaa
     4381 ttccgccccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     4441 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     4501 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     4561 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     4621 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     4681 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     4741 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     4801 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     4861 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     4921 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgag ctttccatgt
     4981 aaatttgtag ccagcttcct tctgattttc aatgtttctt ccaaaggtgc agtctccaaa
     5041 gagattacga atgccttgga aacctggggt gccttgggtc aggacatcaa cttggacatt
     5101 cctagttttc aaatgagtga tgatattgac gatataaaat gggaaaaaac ttcagacaag
     5161 aaaaagattg cacaattcag aaaagagaaa gagactttca aggaaaaaga tacatataag
     5221 ctatttaaaa atggaactct gaaaattaag catctgaaga ccgatgatca ggatatctac
     5281 aaggtatcaa tatatgatac aaaaggaaaa aatgtgttgg aaaaaatatt tgatttgaag
     5341 attcaagaga gggtctcaaa accaaagatc tcctggactt gtatcaacac aaccctgacc
     5401 tgtgaggtaa tgaatggaac tgaccccgaa ttaaacctgt atcaagatgg gaaacatcta
     5461 aaactttctc agagggtcat cacacacaag tggaccacca gcctgagtgc aaaattcaag
     5521 tgcacagcag ggaacaaagt cagcaaggaa tccagtgtcg agcctgtcag ctgtccagag
     5581 aaaggtctgg acatctatct catcattggc atatgtggag gaggcagcct cttgatggtc
     5641 tttgtggcac tgctcgtttt ctatatcacc aaaaggaaaa aacagaggag tcggagaaat
     5701 gatgaggagc tggagacaag agcccacaga gtagctactg aagaaagggg ccggaagccc
     5761 caccaaattc cagcttcaac ccctcagaat taagtcgacc tgcagccaag cttatcgata
     5821 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     5881 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
     5941 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
     6001 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     6061 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     6121 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     6181 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     6241 tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     6301 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     6361 cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga
     6421 gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata
     6481 ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt
     6541 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6601 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6661 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6721 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6781 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6841 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6901 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6961 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7021 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7081 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7141 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7201 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7261 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7321 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7381 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7441 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7501 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7561 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7621 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7681 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7741 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7801 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7861 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7921 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7981 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8041 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8101 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8161 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8221 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8281 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8341 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8401 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8461 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8521 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8581 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8641 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8701 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8761 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8821 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8881 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8941 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     9001 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     9061 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     9121 cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     9181 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     9241 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     9301 ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt
     9361 ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc
     9421 atagataaaa taaaagattt tatttagtct ccagaaaaag ggggg
//
