LOCUS       sequence-436979-        7584 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 7584)
  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 7584)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..7584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     701..718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    835..856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(871..876,877..894,895..901)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    909..925
                     /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             1087..1601
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(1700..2410)
                     /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     2247..2265
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    2422..2974
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2424..2990
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2584..2603)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2813..2830
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     regulatory      3094..3103
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4152..4161
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4175..4591)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4191..4207)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4223..4245)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4658..4999
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             5063..5579
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5779..5798)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5926..5948)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6142..6161)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6261..6283
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6321..6339)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6407..6511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6512..6580,6581..7372)
                     /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(6730..6749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7543..7584,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      721 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      781 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      841 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      901 gtgtggaatt gtgagcggat aacaatttca caattctccc cagctctttc tgcgctgtcc
      961 ataatgaact atttcaggag tattggagta tgtggattca aaacagtggc taattgagtc
     1021 aggattactt aatattcgac tcctaccctc agaatgttgt tctccaactt caagaaactg
     1081 ttacccatga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
     1141 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
     1201 gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt
     1261 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
     1321 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
     1381 gaaactgctg agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc
     1441 aggaactgct taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa
     1501 ccttgatctg aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt
     1561 acttaagcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga
     1621 ctaaataaaa tcttttattt tatcgataag cttggctgca ggtcgacggt atcgataagc
     1681 ttgataattc ttaattaatt tacttgtaca gctcgtccat gccgccggtg gagtggcggc
     1741 cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca
     1801 acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg
     1861 tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc ttgatctcgc
     1921 ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag cccatggttt
     1981 tcttctgcat tacggggccg tctgagggga agttggtgcc gcgcagcttc accttgtaga
     2041 tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga
     2101 agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga
     2161 tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg gacaggatgt
     2221 cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc tgggtgccct
     2281 cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc acggagccct
     2341 ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc tcctcgccct
     2401 tgctcaccat ggttgtggcc atattatcat cgtgtttttc aaaggaaaac cacgtccccg
     2461 tggttcgggg ggcctagacg tttttttaac ctcgactaaa cacatgtaaa gcatgtgcac
     2521 cgaggcccca gatcagatcc catacaatgg ggtaccttct gggcatcctt cagccccttg
     2581 ttgaatacgc ttgaggagag ccatttgact ctttccacaa ctatccaact cacaacgtgg
     2641 cactggggtt gtgccgcctt tgcaggtgta tcttatacac gtggcttttg gccgcagagg
     2701 cacctgtcgc caggtggggg gttccgctgc ctgcaaaggg tcgctacaga cgttgtttgt
     2761 cttcaagaag cttccagagg aactgcttcc ttcacgacat tcaacagacc ttgcattcct
     2821 ttggcgagag gggaaagacc cctaggaatg ctcgtcaaga agacagggcc aggtttccgg
     2881 gccctcacat tgccaaaaga cggcaatatg gtggaaaata acatatagac aaacgcacac
     2941 cggccttatt ccaagcggct tcggccagta acgttagggg ggggggcgga attcttagtc
     3001 atagtagctt agctgaactg ggccgtgggg gtcgtcatca tctccaccgg aaccagaaga
     3061 acccaaaagc agcgtaggaa ggtgtggatc accgccgcca tggccggaat catgactatg
     3121 accgccgcct ccgtctgtca tcaaaggcgg gccctggtca cctcctttgt tttcaacctc
     3181 ttccgtcaat tgtggagggc ctccatcatt tccagcagag tcgctagggc tatgaggcag
     3241 cgggtcatgt gggccattgt catcagtgtt gtcagggtcc tgtgggccat tgtcatcagt
     3301 gttgtcaggg tcctgaggca gcgggtcatg tgggccattg tcatcagtgt tgtcagggtc
     3361 ctgtgggcca ttgtcatcag tgttgtcagg gtcctgtggg ccattgtcag gaccacctcc
     3421 aggtgcgcct aggttttgag agcagagtgg gggtccgtcg ccggctccac tcacgagcag
     3481 gtggtgtctg ccctcgttgg agttagagtc agattcatgg ccagaatcat cggtagcttg
     3541 ttgagggtgc gggagggagt catcgtggtg gtgttcatca ctgtgtcgtt gtccatggta
     3601 atacatccag attaaaatcg ccagaaacag gaggagccaa aggagatcaa ccaatagagt
     3661 ccaccagttt tgttgtagat agagagcaat aatgagcagg atgaggtcta ggaagaaggc
     3721 taggaagaag gccaaaagct gccagatggt ggcaccaagt cgccagagca tctccaataa
     3781 gtagatccag atacctaaga caagtaagca cccgaagatg aacagcacaa ttccaaggaa
     3841 caatgcctgt ccgtgcaaat tccagagagc gatgagcagg agggtgatca tcagtaggag
     3901 tatacaaagg gctccaagtg gacagagaag gtctcttcgg aagataaaga tgatcaaaat
     3961 tataattata agcatgagag caaaggaata gaggacaagg agggctcctc cagtccagtc
     4021 actcataacg atgtacagcc aaaacagtag cgccaagagg aggagaagga gagcaaggcc
     4081 tagggaagag gagagggggg gtcctcgagg gggccgtcgc gggcccggtg ggcccctctc
     4141 aaggtcgtgt tccatggtgg ccagatctaa ttccggcgcc tagagaagga gtgagggctg
     4201 gataaaggga ggatcgaggc ggggtcgaac gaggaggttc aagggggaga gacggggcgg
     4261 atggaggaag aggaggcgga ggcttagggt gtacaaaggg cttgacccag ggaggggggt
     4321 caaaagccaa ggcttcccag gtcacgatgt aggggacctg gtctgggtgt ccatgcgggc
     4381 caggtgaaaa gaccttgatc ttaacctggg tgatgaggtc tcggttaaag gtgccgtctc
     4441 gcggccatcc gacgttaaag gttggccatt ctgcagagca gaaggtaacc caacgtctct
     4501 tcttgacatc taccgactgg ttgtgagcga tccgctcgac atctttccag tgacctaagg
     4561 tcaaacttaa gggagtggta acagtctggc cctaattttc agacaaatac agaaacacag
     4621 tcagacagag acaacacaga acgatgctgc agcgctgcag cagacaagac gcgcggcttc
     4681 ggttccaaac cgaaagcaaa aattcagacg gaggcgggaa ctgttttagg ttctcgtctc
     4741 ctaccagaac cacatatcct gacggggtcg gattccacat cgactccctt cctcaggtcg
     4801 ggccacaaaa acggccccca aagtccctgg gacgtctccc agggttgcgg ccgggtgttc
     4861 agaactcgtc agttccacca cgggtccgcc agatacagag ctagttagct aactagtaca
     4921 gacgcaggcg caaacattag atgccggcac aaacacgcac aaagacagag acagacacga
     4981 aacgaccgct ggccagctta cctcccggcg ggggggtcgg tggtccctgg gcaggggtct
     5041 ccaaatctcg gtggaacctc caaatgaaag acccccgagg tgggcagtca atcaatctga
     5101 ggagaccctc ccaaggatca gcgagtccac gattcggatg caaacagcaa gaggctttat
     5161 tgggaatacg ggtacccggg cgacgcagtc tatcggagga ctggcgcgcc gagtgagggg
     5221 ttgtgggctc ttttattgag ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa
     5281 ctgattggtt agttcaaata aggcacaggg tcatttcagg tccttggggc accctggaaa
     5341 catctgatgg ttctctagaa actgctgagg gcgggaccgc atctggggac catctgttct
     5401 tggccctgag ccggggcagg aactgcttac cacagatatc ctgtttggcc catattctgc
     5461 tgtctctctg ttcctaacct tgatctgaac ttctctattc tcagttatgt attttccatg
     5521 ccttgcaaaa tggcgttact taagctagct tgccaaacct acaggtgggg tctttcattc
     5581 cccccttttt ctggagacta aataaaatct tttattttat ctatggctcg tactctatag
     5641 gcttcagctg gtgatattgt tgagtcaaaa ctagagcctg gaccactgat atcctgtctt
     5701 taacaaattg gactaatcgc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     5761 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     5821 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     5881 gactgttggg cgccatctcc ttgcatgcac cattccttgc gccgtcgttt tacaacgtcg
     5941 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     6001 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     6061 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     6121 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     6181 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6241 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     6301 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     6361 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6421 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6481 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6541 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6601 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6661 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6721 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6781 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6841 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6901 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6961 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7021 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7081 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7141 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7201 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7261 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7321 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7381 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7441 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7501 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7561 gcgcgtaatc tgctgcttgc aaac
//
