LOCUS       sequence-437080-        8033 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EBV latent gene LMP2A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8033)
  AUTHORS   Zhang J, Sommermann T, Li X, Gieselmann L, de la Rosa K, Stecklum M,
            Klein F, Kocks C, Rajewsky K
  TITLE     LMP1 and EBNA2 constitute a minimal set of EBV genes for 
            transformation of human B cells.
  JOURNAL   Front Immunol. 2023 Dec 19;14:1331730. doi: 
            10.3389/fimmu.2023.1331730. eCollection 2023.
  PUBMED    38169736
REFERENCE   2  (bases 1 to 8033)
  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: "Front 
            Immunol."; date: "2023-12-19"; volume: "14:1331730. doi"; pages: " 
            10.3389/fimmu.2023.1331730. eCollection 2023"
COMMENT     Sequence Label: MSCV-LMP2A-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8033
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(213..1004,1005..1073))
                     /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     836..855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1074..1178)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1246..1264
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1302..1324)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1424..1443
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1637..1659
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1787..1806
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2006..2522
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    2586..2927
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             2994..3410
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3340..3362
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3378..3394
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    3431..3455
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4988..5012)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5031..5598
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5048..5600
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5192..5209)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5419..5438
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5612..5614,5615..5617,5618..6331)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5657..5678)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5918..5937)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6265..6286
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             6433..6947
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7109..7125
                     /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(7133..7139,7140..7157,7158..7163))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7178..7199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7316..7333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7487..8033,1..42))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7567..7586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
       61 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
      301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
      361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
      661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
      721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
      781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
      841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
      901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
      961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1201 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1261 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1321 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1381 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     1441 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     1501 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     1561 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     1621 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat
     1681 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     1741 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     1801 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     1861 agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga
     1921 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     1981 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     2041 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     2101 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     2161 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     2221 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     2281 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     2341 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     2401 gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggac
     2461 tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg gggtctttca
     2521 tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc cccccgccgg
     2581 gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg
     2641 gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt atctggcgga
     2701 cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg
     2761 actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc
     2821 cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
     2881 aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca
     2941 tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagggccaga
     3001 ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc
     3061 acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc
     3121 caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg
     3181 ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg
     3241 tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta
     3301 agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga
     3361 ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc ggaattagat
     3421 ctctcgaatc acaagtttgt acaaaaaagc aggctccgcg gccgccccct tcaccatggg
     3481 gtccctagaa atggtgccaa tgggcgcggg tccccctagc cccggcgggg atccggatgg
     3541 gtacgatggc ggaaacaact cccaatatcc atctgcttct ggctcttctg ggaacacccc
     3601 caccccaccg aacgatgagg aacgtgaatc taatgaagag cccccaccgc cttatgagga
     3661 cccatattgg ggcaatggcg accgtcactc ggactatcaa ccactaggaa cccaagatca
     3721 aagtctgtac ttgggattgc aacacgacgg gaatgacggg ctccctcccc ctccctactc
     3781 tccacgggat gactcatctc aacacatata cgaagaagcg ggcagaggaa gtatgaatcc
     3841 agtatgcctg cctgtaattg ttgcgcccta cctcttttgg ctggcggcta ttgccgcctc
     3901 gtgtttcacg gcctcagtta gtaccgttgt gaccgccacc ggcttggccc tctcacttct
     3961 actcttggca gcagtggcca gctcatatgc cgctgcacaa aggaaactgc tgacaccggt
     4021 gacagtgctt actgcggttg tcactttctt tgcaatttgc ctaacatgga ggattgagga
     4081 cccacctttt aattctcttc tgtttgcatt gctggccgca gctggcggac tacaaggcat
     4141 ttacgttctg gtgatgcttg tgctcctgat actagcgtac agaaggagat ggcgccgttt
     4201 gactgtttgt ggcggcatca tgtttttggc atgtgtactt gtcctcatcg tcgacgctgt
     4261 tttgcagctg agtcccctcc ttggagctgt aactgtggtt tccatgacgc tgctgctact
     4321 ggctttcgtc ctctggctct cttcgccagg gggcctaggt actcttggtg cagccctttt
     4381 aacattggca gcagctctgg cactgctagc gtcactgatt ttgggcacac ttaacttgac
     4441 tacaatgttc cttctcatgc tcctatggac acttgtggtt ctcctgattt gctcttcgtg
     4501 ctcttcatgt ccactgagca agatccttct ggcacgactg ttcctatatg ctctcgcact
     4561 cttgttgcta gcctccgcgc taatcgctgg tggcagtatt ttgcaaacaa acttcaagag
     4621 tttaagcagc actgaattta tacccaattt gttctgcatg ttattactga ttgtcgctgg
     4681 catactcttc attcttgcta tcctgaccga atggggcagt ggaaatagaa catacggtcc
     4741 agtttttatg tgcctcggtg gcctgctcac catggtagcc ggcgctgtgt ggctgacggt
     4801 gatgtctaac acgcttttgt ctgcctggat tcttacagca ggattcctga ttttcctcat
     4861 tggctttgcc ctctttgggg tcattagatg ctgccgctac tgctgctact actgccttac
     4921 actggaaagt gaggagcgcc caccgacccc atatcgcaac actgtataag caagggtggg
     4981 cgcgccgacc cagctttctt gtacaaagtg gtgattcgag gttaacgaat tccgcccccc
     5041 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     5101 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     5161 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     5221 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     5281 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     5341 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     5401 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     5461 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     5521 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     5581 tgaaaaacac gatgataata tggccacaac catggtgagc aagggcgagg agctgttcac
     5641 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     5701 gtctggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     5761 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     5821 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     5881 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     5941 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     6001 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     6061 cgtctatatc atggccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca
     6121 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     6181 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     6241 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     6301 cactctcggc atggacgagc tgtacaagta atgaattaat taagaattat caagcttatc
     6361 gataccgtcg acctgcagcc aagcttatcg ataaaataaa agattttatt tagtctccag
     6421 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     6481 ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa
     6541 cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     6601 ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa
     6661 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     6721 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     6781 gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc
     6841 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     6901 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc
     6961 ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa
     7021 ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc
     7081 gcagaaagag ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac
     7141 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7201 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7261 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7321 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7381 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7441 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7501 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7561 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7621 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7681 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7741 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7801 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7861 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7921 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7981 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt ttt
//
