LOCUS       sequence-437048-        7899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EBV latent gene EBNA2 in mammalian cells (EBNA2 coding 
            sequence was optimised for expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7899)
  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 7899)
  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-opEBNA2-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..7899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(501..520)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(648..670)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(864..883)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     983..1005
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1043..1061)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1129..1233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1234..1302,1303..2094)
                     /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     complement(1452..1471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2265..2853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2754..2773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3007..3024
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3141..3162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3177..3182,3183..3200,3201..3207)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3215..3231
                     /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)."
     LTR             3393..3907
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(4006..4716)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     4553..4571
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    4728..5280
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4730..5297
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4890..4909)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     5119..5136
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     regulatory      5805..5814
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(6796..7212)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(6812..6828)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(6844..6866)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    7279..7620
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             join(7684..7899,1..301)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 aactgattgg ttagttcaaa taaggcacag ggtcatttca ggtccttggg gcaccctgga
       61 aacatctgat ggttctctag aaactgctga gggcgggacc gcatctgggg accatctgtt
      121 cttggccctg agccggggca ggaactgctt accacagata tcctgtttgg cccatattct
      181 gctgtctctc tgttcctaac cttgatctga acttctctat tctcagttat gtattttcca
      241 tgccttgcaa aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat
      301 tccccccttt ttctggagac taaataaaat cttttatttt atctatggct cgtactctat
      361 aggcttcagc tggtgatatt gttgagtcaa aactagagcc tggaccactg atatcctgtc
      421 tttaacaaat tggactaatc gcctctacgc cggacgcatc gtggccggca tcaccggcgc
      481 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
      541 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
      601 gggactgttg ggcgccatct ccttgcatgc accattcctt gcgccgtcgt tttacaacgt
      661 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
      721 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      781 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
      841 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
      901 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
      961 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     1021 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     1081 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1141 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1201 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1261 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1321 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1381 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1441 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1501 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1561 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1621 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1681 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1741 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     1801 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     1861 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     1921 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     1981 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2041 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2101 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2161 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2401 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2641 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2701 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2881 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2941 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3001 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3061 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3121 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     3181 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaat
     3241 tctccccagc tctttctgcg ctgtccataa tgaactattt caggagtatt ggagtatgtg
     3301 gattcaaaac agtggctaat tgagtcagga ttacttaata ttcgactcct accctcagaa
     3361 tgttgttctc caacttcaag aaactgttac ccatgaaaga cccccgctga cgggtagtca
     3421 atcactcaga ggagaccctc ccaaggaaca gcgagaccac aagtcggatg caactgcaag
     3481 agggtttatt ggatacacgg gtacccgggc gacgcagtct atcggaggac tggcgcgccg
     3541 agtgaggggt tgtgggctct tttattgagc tcggggagca gaagcgcgcg aacagaagcg
     3601 agaagcgaac tgattggtta gttcaaataa ggcacagggt catttcaggt ccttggggca
     3661 ccctggaaac atctgatggt tctctagaaa ctgctgaggg cgggaccgca tctggggacc
     3721 atctgttctt ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc
     3781 atattctgct gtctctctgt tcctaacctt gatctgaact tctctattct cagttatgta
     3841 ttttccatgc cttgcaaaat ggcgttactt aagctagctt gccaaaccta caggtggggt
     3901 ctttcattcc cccctttttc tggagactaa ataaaatctt ttattttatc gataagcttg
     3961 gctgcaggtc gacggtatcg ataagcttga taattcttaa ttaatttact tgtacagctc
     4021 gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg tactgttcca cgatggtgta
     4081 gtcctcgttg tgggaggtga tgtccaactt gatgttgacg ttgtaggcgc cgggcagctg
     4141 cacgggcttc ttggccttgt aggtggtctt gacctcagcg tcgtagtggc cgccgtcctt
     4201 cagcttcagc ctctgcttga tctcgccctt cagggcgccg tcctcggggt acatccgctc
     4261 ggaggaggcc tcccagccca tggttttctt ctgcattacg gggccgtctg aggggaagtt
     4321 ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc tgcagggagg agtcctgggt
     4381 cacggtcacc acgccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg
     4441 gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc ttcacgtagg ccttggagcc
     4501 gtacatgaac tgaggggaca ggatgtccca ggcgaagggc agggggccac ccttggtcac
     4561 cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc
     4621 gaactcgtgg ccgttcacgg agccctccat gtgcaccttg aagcgcatga actccttgat
     4681 gatggccatg ttatcctcct cgcccttgct caccatggtt gtggccatat tatcatcgtg
     4741 tttttcaaag gaaaaccacg tccccgtggt tcggggggcc tagacgtttt tttaacctcg
     4801 actaaacaca tgtaaagcat gtgcaccgag gccccagatc agatcccata caatggggta
     4861 ccttctgggc atccttcagc cccttgttga atacgcttga ggagagccat ttgactcttt
     4921 ccacaactat ccaactcaca acgtggcact ggggttgtgc cgcctttgca ggtgtatctt
     4981 atacacgtgg cttttggccg cagaggcacc tgtcgccagg tggggggttc cgctgcctgc
     5041 aaagggtcgc tacagacgtt gtttgtcttc aagaagcttc cagaggaact gcttccttca
     5101 cgacattcaa cagaccttgc attcctttgg cgagagggga aagaccccta ggaatgctcg
     5161 tcaagaagac agggccaggt ttccgggccc tcacattgcc aaaagacggc aatatggtgg
     5221 aaaataacat atagacaaac gcacaccggc cttattccaa gcggcttcgg ccagtaacgt
     5281 tagggggggg gggcggaatt cgttaacctc gagttattgg atgctaggtc tgggcctctt
     5341 gctaggtcct tccacgtaat cctcgtcgct gctgggagac tcggttgtct cgaagatgta
     5401 atcccagctc tcgtccaggt cggcagggtc gatgctagga ggataccagt cgtcaggaaa
     5461 caggatagga gcctcggggc tattgtggct ctcaggctcg tggattgggg acacgttagg
     5521 tgtggcggtg tgagagtttc tgcacacagg agcggcgtta gaaggtccag gagtaggaga
     5581 atcgccggca ccttgtcctt gatgcagtcc cagcacaggg ctcagctcag gcatgctagg
     5641 gctagatgtg tttggctcag gcctccaagg gccgccaggt tttctctgct tgtctctgct
     5701 cttgcccttg cctctacctc ttccacgtcc acggcctcta ccacgggact gtcctctgct
     5761 ctgtccctgg gtcttagaag gctgtggagg gtcacagatt ggaggccacc atggaggtcc
     5821 agatggcaga tggtgcagct gctggtcatt gatcacgcca ggaggaggtt gagctggagc
     5881 agcaggaggt ggcaactgac taggtggcag tggcatagga gggatgttgt agaacacggt
     5941 ggggcttcta ggctcagggc tgtcagggtc gttaggggtg ctctggtgtg tcagaggatg
     6001 catagactgg tcgggcacgt gcagcactgg cagatgcatt ctaggtggag atggtgtggg
     6061 ctgcagttct gtaggcctct gcagcacagt cagcagaggt gttggaggaa gtgtggtagg
     6121 cctggttggt cttggaggca ctgtcagtgt agctggaggc agtggtgtag gaggtgtagg
     6181 cctaggaggc atcatcagag gctgaggagg tgtcagtcta ggctggctca gtggagacag
     6241 gatggtagga gcggttggtc tgagtggagg cacatgaggc tgcaccagtc tagcctgtgg
     6301 agctgtctga ggtccctgag gcagaatcag gcctctgtcg cctctagact gccgcacgat
     6361 gtagttggcc atccacagca ttctcatggc agaggccaga ggtccgtgtc cgacatcgta
     6421 tcccagggga tctctatcca gaggactggg ctcttgtgtc caggcatctc ttctttgagg
     6481 cggtggtggt ggtggaggtg gcggcggtgg agaaggtggt ggcggcggag gcggtggcgg
     6541 tggtggcgga ggtggaggcg gaggtggtgg aggcggcggt ggcggtggag gcaatggagg
     6601 tggaacgcct gtgttctcgc ccacgaagat tgtcagaggg atcagaggtg tgtcgctcag
     6661 ctgctcttgg taggggttgc tagggatcac gctcaggcta gggttgccca ggctgtctgt
     6721 atccacgatc aggtggtagg tctgtccgcc gtgcagagcc agatagaagg tgggcatggt
     6781 ggcagatcta attccggcgc ctagagaagg agtgagggct ggataaaggg aggatcgagg
     6841 cggggtcgaa cgaggaggtt caagggggag agacggggcg gatggaggaa gaggaggcgg
     6901 aggcttaggg tgtacaaagg gcttgaccca gggagggggg tcaaaagcca aggcttccca
     6961 ggtcacgatg taggggacct ggtctgggtg tccatgcggg ccaggtgaaa agaccttgat
     7021 cttaacctgg gtgatgaggt ctcggttaaa ggtgccgtct cgcggccatc cgacgttaaa
     7081 ggttggccat tctgcagagc agaaggtaac ccaacgtctc ttcttgacat ctaccgactg
     7141 gttgtgagcg atccgctcga catctttcca gtgacctaag gtcaaactta agggagtggt
     7201 aacagtctgg ccctaatttt cagacaaata cagaaacaca gtcagacaga gacaacacag
     7261 aacgatgctg cagcgctgca gcagacaaga cgcgcggctt cggttccaaa ccgaaagcaa
     7321 aaattcagac ggaggcggga actgttttag gttctcgtct cctaccagaa ccacatatcc
     7381 tgacggggtc ggattccaca tcgactccct tcctcaggtc gggccacaaa aacggccccc
     7441 aaagtccctg ggacgtctcc cagggttgcg gccgggtgtt cagaactcgt cagttccacc
     7501 acgggtccgc cagatacaga gctagttagc taactagtac agacgcaggc gcaaacatta
     7561 gatgccggca caaacacgca caaagacaga gacagacacg aaacgaccgc tggccagctt
     7621 acctcccggc gggggggtcg gtggtccctg ggcaggggtc tccaaatctc ggtggaacct
     7681 ccaaatgaaa gacccccgag gtgggcagtc aatcaatctg aggagaccct cccaaggatc
     7741 agcgagtcca cgattcggat gcaaacagca agaggcttta ttgggaatac gggtacccgg
     7801 gcgacgcagt ctatcggagg actggcgcgc cgagtgaggg gttgtgggct cttttattga
     7861 gctcggggag cagaagcgcg cgaacagaag cgagaagcg
//
