LOCUS       sequence-437068-        8043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EBV latent gene EBNA2 in mammalian cells (EBNA2 coding 
            sequence was optimised for expression and carries a C-terminal 
            3xHA-tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8043)
  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 8043)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..8043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    1425..1449
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2446..2455
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2944..2970
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(2992..3016)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3035..3602
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3052..3604
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3196..3213)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3423..3442
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3616..3618,3619..3621,3622..4335)
                     /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(3661..3682)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3922..3941)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4269..4290
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4437..4951
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5113..5129
                     /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(5137..5143,5144..5161,5162..5167))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5182..5203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5320..5337)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5491..6079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5571..5590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6250..7041,7042..7110))
                     /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     6873..6892
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7111..7215)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7283..7301
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7339..7361)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7461..7480
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7674..7696
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7824..7843
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8043,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctctcg aatcacaagt ttgtacaaaa
     1441 aagcaggctc cgcggccgcc cccttcgcgc ccacccttca ccatgcccac cttctatctg
     1501 gctctgcacg gcggacagac ctaccacctg atcgtggata cagacagcct gggcaaccct
     1561 agcctgagcg tgatccctag caacccctac caagagcagc tgagcgacac acctctgatc
     1621 cctctgacaa tcttcgtggg cgagaacaca ggcgttccac ctccattgcc tccaccgcca
     1681 ccgccgcctc caccacctcc gcctccacct ccgccaccac cgccaccgcc tccgccgcca
     1741 ccaccttctc caccgccgcc acctccacca ccaccaccgc ctcaaagaag agatgcctgg
     1801 acacaagagc ccagtcctct ggatagagat cccctgggat acgatgtcgg acacggacct
     1861 ctggcctctg ccatgagaat gctgtggatg gccaactaca tcgtgcggca gtctagaggc
     1921 gacagaggcc tgattctgcc tcagggacct cagacagctc cacaggctag actggtgcag
     1981 cctcatgtgc ctccactcag accaaccgct cctaccatcc tgtctccact gagccagcct
     2041 agactgacac ctcctcagcc tctgatgatg cctcctaggc ctacacctcc tacaccactg
     2101 cctccagcta cactgacagt gcctccaaga ccaaccaggc ctaccacact tcctccaaca
     2161 cctctgctga ctgtgctgca gaggcctaca gaactgcagc ccacaccatc tccacctaga
     2221 atgcatctgc cagtgctgca cgtgcccgac cagtctatgc atcctctgac acaccagagc
     2281 acccctaacg accctgacag ccctgagcct agaagcccca ccgtgttcta caacatccct
     2341 cctatgccac tgccacctag tcagttgcca cctcctgctg ctccagctca acctcctcct
     2401 ggcgtgatca atgaccagca gctgcaccat ctgccatctg gacctccatg gtggcctcca
     2461 atctgtgacc ctccacagcc ttctaagacc cagggacaga gcagaggaca gtcccgtggt
     2521 agaggccgtg gacgtggaag aggtagaggc aagggcaaga gcagagacaa gcagagaaaa
     2581 cctggcggcc cttggaggcc tgagccaaac acatctagcc ctagcatgcc tgagctgagc
     2641 cctgtgctgg gactgcatca aggacaaggt gccggcgatt ctcctactcc tggaccttct
     2701 aacgccgctc ctgtgtgcag aaactctcac accgccacac ctaacgtgtc cccaatccac
     2761 gagcctgaga gccacaatag ccccgaggct cctatcctgt ttcctgacga ctggtatcct
     2821 cctagcatcg accctgccga cctggacgag agctgggatt acatcttcga gacaaccgag
     2881 tctcccagca gcgacgagga ttacgtggaa ggacctagca agaggcccag acctagcatc
     2941 caatacccat acgatgttcc agattacgct taagcaaggg tgggcgcgcc gacccagctt
     3001 tcttgtacaa agtggtgatt cgaggttaac gaattccgcc ccccccccct aacgttactg
     3061 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     3121 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     3181 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     3241 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     3301 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     3361 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     3421 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     3481 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     3541 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     3601 aatatggcca caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3661 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtctgg cgagggcgag
     3721 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3781 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     3841 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3901 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3961 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4021 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4081 gacaagcaga agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc
     4141 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4201 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     4261 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4321 gagctgtaca agtaatgaat taattaagaa ttatcaagct tatcgatacc gtcgacctgc
     4381 agccaagctt atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     4441 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     4501 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     4561 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4621 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     4681 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     4741 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     4801 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac
     4861 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     4921 gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca
     4981 acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga
     5041 atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg
     5101 agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5161 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5221 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5281 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5341 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5401 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5461 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5521 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5581 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5641 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5701 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5761 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5821 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5881 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5941 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6001 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6061 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6121 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6301 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6361 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6421 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6481 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6541 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6601 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6661 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6721 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6781 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6841 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6901 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6961 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7021 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7081 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7141 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7201 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7261 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     7321 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     7381 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     7441 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     7501 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     7561 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     7621 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7681 acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     7741 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     7801 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     7861 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt
     7921 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     7981 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     8041 gga
//
