LOCUS       sequence-270832-        8709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of multiple human genes (BCL2 and MYC).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8709)
  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 8709)
  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-MYC-t2A-BCL2
FEATURES             Location/Qualifiers
     source          1..8709
                     /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             1478..2794
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPIPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     CDS             2816..2869
                     /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    3626..4193
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3643..4195
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3787..3804)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4014..4033
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             5104..5618
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5780..5796
                     /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(5804..5810,5811..5828,5829..5834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5849..5870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5987..6004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6158..6746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6238..6257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6917..7708,7709..7777))
                     /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     7540..7559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7778..7882)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7950..7968
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8006..8028)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8128..8147
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8341..8363
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8491..8510
                     /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 ccatggattt
     1441 ttttcgggta gtggaaaacc agcagcctcc cgcgacgatg cccctcaacg ttagcttcac
     1501 caacaggaac tatgacctcg actacgactc ggtgcagccg tatttctact gcgacgagga
     1561 ggagaacttc taccagcagc agcagcagag cgagctgcag cccccggcgc ccagcgagga
     1621 tatctggaag aaattcgagc tgctgcccat cccgcccctg tcccctagcc gccgctccgg
     1681 gctctgctcg ccctcctacg ttgcggtcac acccttctcc cttcggggag acaacgacgg
     1741 cggtggcggg agcttctcca cggccgacca gctggagatg gtgaccgagc tgctgggagg
     1801 agacatggtg aaccagagtt tcatctgcga cccggacgac gagaccttca tcaaaaacat
     1861 catcatccag gactgtatgt ggagcggctt ctcggccgcc gccaagctcg tctcagagaa
     1921 gctggcctcc taccaggctg cgcgcaaaga cagcggcagc ccgaaccccg cccgcggcca
     1981 cagcgtctgc tccacctcca gcttgtacct gcaggatctg agcgccgccg cctcagagtg
     2041 catcgacccc tcggtggtct tcccctaccc tctcaacgac agcagctcgc ccaagtcctg
     2101 cgcctcgcaa gactccagcg ccttctctcc gtcctcggat tctctgctct cctcgacgga
     2161 gtcctccccg cagggcagcc ccgagcccct ggtgctccat gaggagacac cgcccaccac
     2221 cagcagcgac tctgaggagg aacaagaaga tgaggaagaa atcgatgttg tttctgtgga
     2281 aaagaggcag gctcctggca aaaggtcaga gtctggatca ccttctgctg gaggccacag
     2341 caaacctcct cacagcccac tggtcctcaa gaggtgccac gtctccacac atcagcacaa
     2401 ctacgcagcg cctccctcca ctcggaagga ctatcctgct gccaagaggg tcaagttgga
     2461 cagtgtcaga gtcctgagac agatcagcaa caaccgaaaa tgcaccagcc ccaggtcctc
     2521 ggacaccgag gagaatgtca agaggcgaac acacaacgtc ttggagcgcc agaggaggaa
     2581 cgagctaaaa cggagctttt ttgccctgcg tgaccagatc ccggagttgg aaaacaatga
     2641 aaaggccccc aaggtagtta tccttaaaaa agccacagca tacatcctgt ccgtccaagc
     2701 agaggagcaa aagctcattt ctgaagagga cttgttgcgg aaacgacgag aacagttgaa
     2761 acacaaactt gaacagctac ggaactcttg tgcgaacgga tccggcagcg gcggggaagg
     2821 aaggggctcc ctgctcacct gcggcgacgt ggaagagaac cccggaccta tggcgcacgc
     2881 tgggagaaca gggtacgata accgggagat agtgatgaag tacatccatt ataagctgtc
     2941 gcagaggggc tacgagtggg atgcgggaga tgtgggagcc gctcctcctg gtgccgctcc
     3001 cgcaccgggc atcttctcct cccagcctgg gcacacgcct catccagccg catcccggga
     3061 tcctgtcgcc aggacctcgc ctctgcagac accggctgct cctggagctg ctgcaggacc
     3121 tgcgctcagc ccggtgccac ctgtggtcca cctgaccctc cgccaggccg gcgacgactt
     3181 ctcccgccgc taccgccgcg acttcgccga gatgtccagc cagctgcacc tgacgccctt
     3241 caccgcgcgg ggacgctttg ccacggtggt ggaggagctc ttcagggacg gggtgaactg
     3301 ggggaggatt gtggccttct ttgagttcgg tggggtcatg tgtgtggaga gcgtcaaccg
     3361 ggagatgtcg cccctggtgg acaacatcgc cctgtggatg actgagtacc tgaaccggca
     3421 cctgcacacc tggatccagg ataacggagg ctgggatgcc tttgtggaac tgtacggccc
     3481 cagcatgcgg cctctgtttg atttctcctg gctgtctctg aagactctgc tcagtttggc
     3541 cctggtggga gcttgcatca ccctgggtgc ctatctgggc cacaagtgat agaacgcggc
     3601 cgcaccggtg gatcctacgt agaattccgc cccccccccc taacgttact ggccgaagcc
     3661 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     3721 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     3781 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     3841 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     3901 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     3961 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     4021 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     4081 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     4141 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     4201 acaaccatgg tgagctttcc atgtaaattt gtagccagct tccttctgat tttcaatgtt
     4261 tcttccaaag gtgcagtctc caaagagatt acgaatgcct tggaaacctg gggtgccttg
     4321 ggtcaggaca tcaacttgga cattcctagt tttcaaatga gtgatgatat tgacgatata
     4381 aaatgggaaa aaacttcaga caagaaaaag attgcacaat tcagaaaaga gaaagagact
     4441 ttcaaggaaa aagatacata taagctattt aaaaatggaa ctctgaaaat taagcatctg
     4501 aagaccgatg atcaggatat ctacaaggta tcaatatatg atacaaaagg aaaaaatgtg
     4561 ttggaaaaaa tatttgattt gaagattcaa gagagggtct caaaaccaaa gatctcctgg
     4621 acttgtatca acacaaccct gacctgtgag gtaatgaatg gaactgaccc cgaattaaac
     4681 ctgtatcaag atgggaaaca tctaaaactt tctcagaggg tcatcacaca caagtggacc
     4741 accagcctga gtgcaaaatt caagtgcaca gcagggaaca aagtcagcaa ggaatccagt
     4801 gtcgagcctg tcagctgtcc agagaaaggt ctggacatct atctcatcat tggcatatgt
     4861 ggaggaggca gcctcttgat ggtctttgtg gcactgctcg ttttctatat caccaaaagg
     4921 aaaaaacaga ggagtcggag aaatgatgag gagctggaga caagagccca cagagtagct
     4981 actgaagaaa ggggccggaa gccccaccaa attccagctt caacccctca gaattaagtc
     5041 gacctgcagc caagcttatc gataaaataa aagattttat ttagtctcca gaaaaagggg
     5101 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     5161 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     5221 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     5281 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     5341 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     5401 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     5461 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc
     5521 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     5581 tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg
     5641 gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact
     5701 gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga
     5761 gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5821 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5881 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5941 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6001 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6061 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7021 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7921 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     7981 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     8041 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     8101 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     8161 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     8221 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     8281 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     8341 cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg agatggcgcc
     8401 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     8461 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     8521 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc gattagtcca
     8581 atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac aatatcacca
     8641 gctgaagcct atagagtacg agccatagat aaaataaaag attttattta gtctccagaa
     8701 aaagggggg
//
