LOCUS       sequence-270005-        9114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides episomal replication of the destination vector in primate 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9114)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 9114)
  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"
COMMENT     Sequence Label: pDEST-EB-R4R3
FEATURES             Location/Qualifiers
     source          1..9114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..168
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     152..168
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    188..311
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(659..964)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(761..780)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1284..1967)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     1882..1901
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1968..2070)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2216..2339)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2347..2363)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2827..4616
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     CDS             complement(4918..6843)
                     /codon_start=1
                     /product="Epstein-Barr nuclear antigen 1, also known as 
                     EBNA-1"
                     /label=EBNA1
                     /note="crucial for latent viral infection, and for episomal
                     amplification of vectors containing the oriP origin (Okita 
                     et al., 2013)"
                     /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR
                     GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG
                     AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGGAGGAGAGGGA
                     GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG
                     GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGGAGAG
                     GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG
                     GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP
                     PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR
                     RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL
                     YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK
                     DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE
                     GEEGQE"
     primer_bind     complement(7278..7296)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7364..7468
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7469..7537,7538..8329)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7687..7706)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8500..9088
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8989..9008
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcctggcc ttttgctggc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga
       61 cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg
      121 gaaggccatc cagcctcgcg tcgcggctag tcaggaaaca gctatgacca tgattacgcc
      181 aagctatcaa ctttgtatag aaaagttgaa cgagaaacgt aaaatgatat aaatatcaat
      241 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
      301 cagtcactat ggtcgaccga cagccttcca aatgttcttc tcaaacggaa tcgtcgtatc
      361 cagcctactc gctattgtcc tcaatgccgt attaaatcat aaaaagaaat aagaaaaaga
      421 ggtgcgagcc tcttttttgt gtgacaaaat aaaaacatct acctattcat atacgctagt
      481 gtcatagtcc tgaaaatcat ctgcatcaag aacaatttca caactcttat acttttctct
      541 tacaagtcgt tcggcttcat ctggattttc agcctctata cttactaaac gtgataaagt
      601 ttctgtaatt tctactgtat cgacctgcag actggctgtg tataagggag cctgacattt
      661 atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat
      721 cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc
      781 gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca
      841 gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca ataatatcac
      901 tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact
      961 gcatttcacc agcccctgtt ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg
     1021 acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag acgacgggct
     1081 tcattctgca tggttgtgct taccagaccg gagatattga catcatatat gccttgagca
     1141 actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca tacctctttt
     1201 tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat cagcgcgcaa
     1261 atacgcatac tgttatctgg cttttagtaa gccggatcca cgcggcgttt acgccccccc
     1321 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     1381 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     1441 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     1501 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     1561 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     1621 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     1681 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     1741 gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     1801 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     1861 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     1921 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     1981 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     2041 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag
     2101 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc
     2161 acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttacg gccgccatag
     2221 tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca aaatctaatt
     2281 taatatattg atatttatat cattttacgt ttctcgttca actttattat acatagttga
     2341 taattcactg gccgtcgttt tacctcgact ctagaggatc gatgccccgc cccggacgaa
     2401 ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt aatcccttca
     2461 gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg gggggaccaa
     2521 acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc acatttgcca
     2581 acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca acacaacatt
     2641 gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac atgtacctcc
     2701 caggggccca ggaagactac gggaggctac accaacgtca atcagagggg cctgtgtagc
     2761 taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc ccccttgtta
     2821 accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag actaacccta
     2881 attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg ttagtaaaag
     2941 ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta tttacatggg
     3001 gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa
     3061 ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct tcgtttagct
     3121 aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt
     3181 ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc attgtgctat
     3241 gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag gaatgaaaca
     3301 ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa agactggatg
     3361 tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa tatgatactg
     3421 gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt gttacactct
     3481 atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact ggttgtctct
     3541 aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca aagacaagtg
     3601 gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac acgccgccct
     3661 gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc ccgctaacca
     3721 ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat acctgcataa
     3781 gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca aattacacac
     3841 acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag gtcgctgaga
     3901 gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg gatagcatat
     3961 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4021 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     4081 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4141 gctatcctaa tctgtatccg ggtagcatat gctatcctaa tagagattag ggtagtatat
     4201 gctatcctaa tttatatctg ggtagcatat actacccaaa tatctggata gcatatgcta
     4261 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gcataggcta
     4321 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     4381 tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     4441 tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta
     4501 tcctcatgca tatacagtca gcatatgata cccagtagta gagtgggagt gctatccttt
     4561 gcatatgccg ccacctccca agggggcgtg aattttcgct gcttgtcctt ttcctgctgg
     4621 ttgctcccat tcttaggtga atttaaggag gccaggctaa agccgtcgca tgtctgattg
     4681 ctcaccaggt aaatgtcgct aatgttttcc aacgcgagaa ggtgttgagc gcggagctga
     4741 gtgacgtgac aacatgggta tgcccaattg ccccatgttg ggaggacgaa aatggtgaca
     4801 agacagatgg ccagaaatac accaacagca cgcatgatgt ctactgggga tttattcttt
     4861 agtgcggggg aatacacggc ttttaatacg attgagggcg tctcctaaca agttacatca
     4921 ctcctgccct tcctcaccct catctccatc acctccttca tctccgtcat ctccgtcatc
     4981 accctccgcg gcagcccctt ccaccatagg tggaaaccag ggaggcaaat ctactccatc
     5041 gtcaaagctg cacacagtca ccctgatatt gcaggtagga gcgggctttg tcataacaag
     5101 gtccttaatc gcatccttca aaacctcagc aaatatatga gtttgtaaaa agaccatgaa
     5161 ataacagaca atggactccc ttagcgggcc aggttgtggg ccgggtccag gggccattcc
     5221 aaaggggaga cgactcaatg gtgtaagacg acattgtgga atagcaaggg cagttcctcg
     5281 ccttaggttg taaagggagg tcttactacc tccatatacg aacacaccgg cgacccaagt
     5341 tccttcgtcg gtagtccttt ctacgtgact cctagccagg agagctctta aaccttctgc
     5401 aatgttctca aatttcgggt tggaacctcc ttgaccacga tgctttccaa accaccctcc
     5461 ttttttgcgc ctgcctccat caccctgacc ccggggtcca gtgcttgggc cttctcctgg
     5521 gtcatctgcg gggccctgct ctatcgctcc cgggggcacg tcaggctcac catctgggcc
     5581 accttcttgg tggtattcaa aataatcggc ttcccctaca gggtggaaaa atggccttct
     5641 acctggaggg ggcctgcgcg gtggagaccc ggatgatgat gactgactac tgggactcct
     5701 gggcctcttt tctccacgtc cacgacctct ccccctggct ctttcacgac ttccccccct
     5761 ggctctttca cgtcctctac cccggcggcc tccactacct cctcgacccc ggcctccact
     5821 acctcctcga ccccggcctc cactgcctcc tcgaccccgg cctccacctc ctgctcctgc
     5881 ccctcctgct cctgcccctc ctcctgctcc tgcccctcct gcccctcctg ctcctgcccc
     5941 tcctgcccct cctgctcctg cccctcctgc ccctcctgct cctgcccctc ctgcccctcc
     6001 tcctgctcct gcccctcctg cccctcctcc tgctcctgcc cctcctgccc ctcctgctcc
     6061 tgcccctcct gcccctcctg ctcctgcccc tcctgcccct cctgctcctg cccctcctgc
     6121 tcctgcccct cctgctcctg cccctcctgc tcctgcccct cctgcccctc ctgcccctcc
     6181 tcctgctcct gcccctcctg ctcctgcccc tcctgcccct cctgcccctc ctgctcctgc
     6241 ccctcctcct gctcctgccc ctcctgcccc tcctgcccct cctcctgctc ctgcccctcc
     6301 tgcccctcct cctgctcctg cccctcctcc tgctcctgcc cctcctgccc ctcctgcccc
     6361 tcctcctgct cctgcccctc ctgcccctcc tcctgctcct gcccctcctc ctgctcctgc
     6421 ccctcctgcc cctcctgccc ctcctcctgc tcctgcccct cctcctgctc ctgcccctcc
     6481 tgcccctcct gcccctcctg cccctcctcc tgctcctgcc cctcctcctg ctcctgcccc
     6541 tcctgctcct gcccctcccg ctcctgctcc tgctcctgtt ccaccgtggg tccctttgca
     6601 gccaatgcaa cttggacgtt tttggggtct ccggacacca tctctatgtc ttggccctga
     6661 tcctgagccg cccggggctc ctggtcttcc gcctcctcgt cctcgtcctc ttccccgtcc
     6721 tcgtccatgg ttatcacccc ctcttctttg aggtccactg ccgccggagc cttctggtcc
     6781 agatgtgtct cccttctctc ctaggccatt tccaggtcct gtacctggcc cctcgtcaga
     6841 catgattcac actaaaagag atcaatagac atctttatta gacgacgctc agtgaataca
     6901 gggagtgcag actcctgccc cctccaacag cccccccacc ctcatcccct tcatggtcgc
     6961 tgtcagacag atccaggtct gaaaattccc catcctccga accatcctcg tcctcatcac
     7021 caattactcg cagcccggaa aactcccgct gaacatcctc aagatttgcg tcctgagcct
     7081 caagccaggc ctcaaattcc tcgtccccct ttttgctgga cggtagggat ggggattctc
     7141 gggacccctc ctcttcctct tcaaggtcac cagacagaga tgctactggg gcaacggaag
     7201 aaaagctggg tgcggcctgt gaggatcagc ttatcgatga taagctgtca aacatgagaa
     7261 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     7321 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7381 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7441 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7501 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7561 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7621 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7681 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     7741 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7801 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7861 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7921 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7981 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     8041 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     8101 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     8161 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     8221 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     8281 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     8341 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     8401 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     8461 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     8521 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     8581 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     8641 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     8701 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     8761 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     8821 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     8881 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     8941 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     9001 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     9061 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacg
//
