LOCUS       sequence-437076-        7700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EBV latent gene LMP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7700)
  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 7700)
  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-LMP1-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..7700
                     /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"
     regulatory      4528..4537
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4655..4679)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4698..5265
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4715..5267
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4859..4876)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5086..5105
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5279..5281,5282..5284,5285..5998)
                     /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(5324..5345)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5585..5604)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5932..5953
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             6100..6614
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6776..6792
                     /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(6800..6806,6807..6824,6825..6830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6845..6866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6983..7000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7154..7700,1..42))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7234..7253)
                     /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 tcaccatgga
     3481 acacgacctt gagaggggcc caccgggccc gcgacggccc cctcgaggac cccccctctc
     3541 ctcttcccta ggccttgctc tccttctcct cctcttggcg ctactgtttt ggctgtacat
     3601 cgttatgagt gactggactg gaggagccct ccttgtcctc tattcctttg ctctcatgct
     3661 tataattata attttgatca tctttatctt ccgaagagac cttctctgtc cacttggagc
     3721 cctttgtata ctcctactga tgatcaccct cctgctcatc gctctctgga atttgcacgg
     3781 acaggcattg ttccttggaa ttgtgctgtt catcttcggg tgcttacttg tcttaggtat
     3841 ctggatctac ttattggaga tgctctggcg acttggtgcc accatctggc agcttttggc
     3901 cttcttccta gccttcttcc tagacctcat cctgctcatt attgctctct atctacaaca
     3961 aaactggtgg actctattgg ttgatctcct ttggctcctc ctgtttctgg cgattttaat
     4021 ctggatgtat taccatggac aacgacacag tgatgaacac caccacgatg actccctccc
     4081 gcaccctcaa caagctaccg atgattctgg ccatgaatct gactctaact ccaacgaggg
     4141 cagacaccac ctgctcgtga gtggagccgg cgacggaccc ccactctgct ctcaaaacct
     4201 aggcgcacct ggaggtggtc ctgacaatgg cccacaggac cctgacaaca ctgatgacaa
     4261 tggcccacag gaccctgaca acactgatga caatggccca catgacccgc tgcctcagga
     4321 ccctgacaac actgatgaca atggcccaca ggaccctgac aacactgatg acaatggccc
     4381 acatgacccg ctgcctcata gccctagcga ctctgctgga aatgatggag gccctccaca
     4441 attgacggaa gaggttgaaa acaaaggagg tgaccagggc ccgcctttga tgacagacgg
     4501 aggcggcggt catagtcatg attccggcca tggcggcggt gatccacacc ttcctacgct
     4561 gcttttgggt tcttctggtt ccggtggaga tgatgacgac ccccacggcc cagttcagct
     4621 aagctactat gactaagcaa gggtgggcgc gccgacccag ctttcttgta caaagtggtg
     4681 attcgaggtt aacgaattcc gccccccccc cctaacgtta ctggccgaag ccgcttggaa
     4741 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     4801 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     4861 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     4921 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     4981 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     5041 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     5101 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     5161 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     5221 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     5281 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     5341 cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg
     5401 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     5461 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     5521 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     5581 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     5641 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     5701 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     5761 catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     5821 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     5881 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     5941 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg
     6001 aattaattaa gaattatcaa gcttatcgat accgtcgacc tgcagccaag cttatcgata
     6061 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     6121 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
     6181 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
     6241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     6301 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     6361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     6421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     6481 tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     6541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     6601 cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga
     6661 gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata
     6721 ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt
     6781 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6841 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6901 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6961 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7021 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7081 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7321 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7561 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7681 ctggtagcgg tggttttttt
//
