LOCUS       sequence-437073-        7928 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 7928)
  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 7928)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..7928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             141..657
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    721..1062
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1129..1545
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1475..1497
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1513..1529
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1558..1567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3074..3640
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3090..3642
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3234..3251)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3461..3480
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3654..4364
                     /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     complement(3799..3817)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     LTR             4463..4977
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5139..5155
                     /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(5163..5169,5170..5187,5188..5193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5208..5229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5346..5363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5517..6105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5597..5616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6276..7067,7068..7136))
                     /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     6899..6918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7137..7241)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7309..7327
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7365..7387)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7487..7506
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7700..7722
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7850..7869
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa
       61 caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt
      121 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
      181 gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa
      241 ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc
      301 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt
      361 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
      421 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
      481 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc
      541 ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggactcgct
      601 gatccttggg agggtctcct cagattgatt gactgcccac ctcgggggtc tttcatttgg
      661 aggttccacc gagatttgga gacccctgcc cagggaccac cgaccccccc gccgggaggt
      721 aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc
      781 taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt
      841 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt
      901 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca
      961 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt
     1021 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt
     1081 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt
     1141 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
     1201 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
     1261 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
     1321 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
     1381 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
     1441 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
     1501 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat tagatctgcc
     1561 accatggggt ccctagaaat ggtgccaatg ggcgcgggtc cccctagccc cggcggggat
     1621 ccggatgggt acgatggcgg aaacaactcc caatatccat ctgcttctgg ctcttctggg
     1681 aacaccccca ccccaccgaa cgatgaggaa cgtgaatcta atgaagagcc cccaccgcct
     1741 tatgaggacc catattgggg caatggcgac cgtcactcgg actatcaacc actaggaacc
     1801 caagatcaaa gtctgtactt gggattgcaa cacgacggga atgacgggct ccctccccct
     1861 ccctactctc cacgggatga ctcatctcaa cacatatacg aagaagcggg cagaggaagt
     1921 atgaatccag tatgcctgcc tgtaattgtt gcgccctacc tcttttggct ggcggctatt
     1981 gccgcctcgt gtttcacggc ctcagttagt accgttgtga ccgccaccgg cttggccctc
     2041 tcacttctac tcttggcagc agtggccagc tcatatgccg ctgcacaaag gaaactgctg
     2101 acaccggtga cagtgcttac tgcggttgtc actttctttg caatttgcct aacatggagg
     2161 attgaggacc caccttttaa ttctcttctg tttgcattgc tggccgcagc tggcggacta
     2221 caaggcattt acgttctggt gatgcttgtg ctcctgatac tagcgtacag aaggagatgg
     2281 cgccgtttga ctgtttgtgg cggcatcatg tttttggcat gtgtacttgt cctcatcgtc
     2341 gacgctgttt tgcagctgag tcccctcctt ggagctgtaa ctgtggtttc catgacgctg
     2401 ctgctactgg ctttcgtcct ctggctctct tcgccagggg gcctaggtac tcttggtgca
     2461 gcccttttaa cattggcagc agctctggca ctgctagcgt cactgatttt gggcacactt
     2521 aacttgacta caatgttcct tctcatgctc ctatggacac ttgtggttct cctgatttgc
     2581 tcttcgtgct cttcatgtcc actgagcaag atccttctgg cacgactgtt cctatatgct
     2641 ctcgcactct tgttgctagc ctccgcgcta atcgctggtg gcagtatttt gcaaacaaac
     2701 ttcaagagtt taagcagcac tgaatttata cccaatttgt tctgcatgtt attactgatt
     2761 gtcgctggca tactcttcat tcttgctatc ctgaccgaat ggggcagtgg aaatagaaca
     2821 tacggtccag tttttatgtg cctcggtggc ctgctcacca tggtagccgg cgctgtgtgg
     2881 ctgacggtga tgtctaacac gcttttgtct gcctggattc ttacagcagg attcctgatt
     2941 ttcctcattg gctttgccct ctttggggtc attagatgct gccgctactg ctgctactac
     3001 tgccttacac tggaaagtga ggagcgccca ccgaccccat atcgcaacac tgtataactc
     3061 gaggttaacg aattccgccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     3121 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     3181 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     3241 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     3301 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     3361 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     3421 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     3481 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     3541 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     3601 ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga
     3661 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     3721 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     3781 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     3841 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     3901 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     3961 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     4021 tcatctacaa ggtgaagctg cgcggcacca acttcccctc agacggcccc gtaatgcaga
     4081 agaaaaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     4141 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     4201 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     4261 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     4321 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaaattaat taagaattat
     4381 caagcttatc gataccgtcg acctgcagcc aagcttatcg ataaaataaa agattttatt
     4441 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     4501 agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca
     4561 aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
     4621 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt
     4681 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     4741 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     4801 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg
     4861 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     4921 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg
     4981 taacagtttc ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat
     5041 cctgactcaa ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat
     5101 tatggacagc gcagaaagag ctggggagaa ttgtgaaatt gttatccgct cacaattcca
     5161 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5221 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5341 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5401 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5461 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5521 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5581 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5641 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5701 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5761 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5821 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5881 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5941 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6001 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6061 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6121 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6181 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6241 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6301 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6361 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6421 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6481 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6541 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6601 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6661 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6721 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6781 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6841 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6901 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6961 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7021 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7081 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7141 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7201 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7261 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7321 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     7381 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     7441 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     7501 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     7561 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7621 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     7681 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg
     7741 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     7801 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     7861 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     7921 cgtagagg
//
