LOCUS       Exported                9113 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Provides episomal replication of the destination vector in primate 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    pDEST-EB-R4R3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9113)
  TITLE     Minoru Ko Lab plasmids
REFERENCE   2  (bases 1 to 9113)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9113
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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    187..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(1314..1967)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
     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      2826..4615
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     CDS             complement(4917..6842)
                     /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
                     AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGG?GGAG?GGG?
                     GG?GGAG?GGGAGAGGGAGGAGAGGG?GGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG
                     G?GG?GG?GAGGAGAGGAGAGGAGAGGAGGAGAGGAGG?G?GG?GG?G?GGGAGGAGAG
                     GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG
                     GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP
                     PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR
                     RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL
                     YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK
                     DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE
                     GEEGQE"
     primer_bind     complement(7277..7295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7363..7467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7468..8328
                     /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(7686..7705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8499..9087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8988..9007
                     /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 gatccccgcc ccggacgaac
     2401 taaacctgac tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag
     2461 ttggttggta caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa
     2521 cacaaagggg ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa
     2581 cactgagtgg ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg
     2641 cctttatgtg taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc
     2701 aggggcccag gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct
     2761 accgataagc ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa
     2821 ccctaaacgg gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa
     2881 ttcaatagca tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg
     2941 gtcctaagga acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg
     3001 tcaggattcc acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag
     3061 gagcgggcag tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta
     3121 atagaataac tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt
     3181 tcaggtgacg cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg
     3241 acaccaatat aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat
     3301 tctgaatatc tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt
     3361 ccatctcaca cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg
     3421 ggttattaag atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta
     3481 tttgtaacaa ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta
     3541 acacccccga aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg
     3601 ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg
     3661 cggttttgga ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac
     3721 tgcggtcaaa ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag
     3781 taggtgggcg ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca
     3841 cttgcgcctg agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag
     3901 cacggtgggc taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg
     3961 ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     4021 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg
     4081 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     4141 ctatcctaat ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg
     4201 ctatcctaat ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat
     4261 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag cataggctat
     4321 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4381 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4441 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     4501 cctcatgcat atacagtcag catatgatac ccagtagtag agtgggagtg ctatcctttg
     4561 catatgccgc cacctcccaa gggggcgtga attttcgctg cttgtccttt tcctgctggt
     4621 tgctcccatt cttaggtgaa tttaaggagg ccaggctaaa gccgtcgcat gtctgattgc
     4681 tcaccaggta aatgtcgcta atgttttcca acgcgagaag gtgttgagcg cggagctgag
     4741 tgacgtgaca acatgggtat gcccaattgc cccatgttgg gaggacgaaa atggtgacaa
     4801 gacagatggc cagaaataca ccaacagcac gcatgatgtc tactggggat ttattcttta
     4861 gtgcggggga atacacggct tttaatacga ttgagggcgt ctcctaacaa gttacatcac
     4921 tcctgccctt cctcaccctc atctccatca cctccttcat ctccgtcatc tccgtcatca
     4981 ccctccgcgg cagccccttc caccataggt ggaaaccagg gaggcaaatc tactccatcg
     5041 tcaaagctgc acacagtcac cctgatattg caggtaggag cgggctttgt cataacaagg
     5101 tccttaatcg catccttcaa aacctcagca aatatatgag tttgtaaaaa gaccatgaaa
     5161 taacagacaa tggactccct tagcgggcca ggttgtgggc cgggtccagg ggccattcca
     5221 aaggggagac gactcaatgg tgtaagacga cattgtggaa tagcaagggc agttcctcgc
     5281 cttaggttgt aaagggaggt cttactacct ccatatacga acacaccggc gacccaagtt
     5341 ccttcgtcgg tagtcctttc tacgtgactc ctagccagga gagctcttaa accttctgca
     5401 atgttctcaa atttcgggtt ggaacctcct tgaccacgat gctttccaaa ccaccctcct
     5461 tttttgcgcc tgcctccatc accctgaccc cggggtccag tgcttgggcc ttctcctggg
     5521 tcatctgcgg ggccctgctc tatcgctccc gggggcacgt caggctcacc atctgggcca
     5581 ccttcttggt ggtattcaaa ataatcggct tcccctacag ggtggaaaaa tggccttcta
     5641 cctggagggg gcctgcgcgg tggagacccg gatgatgatg actgactact gggactcctg
     5701 ggcctctttt ctccacgtcc acgacctctc cccctggctc tttcacgact tccccccctg
     5761 gctctttcac gtcctctacc ccggcggcct ccactacctc ctcgaccccg gcctccacta
     5821 cctcctcgac cccggcctcc actgcctcct cgaccccggc ctccacctcc tgctcctgcc
     5881 cctcctgctc ctgcccctcc tcctgctcct gcccctcctg cccctcctgc tcctgcccct
     5941 cctgcccctc ctgctcctgc ccctcctgcc cctcctgctc ctgcccctcc tgcccctcct
     6001 cctgctcctg cccctcctgc ccctcctcct sctcctsccc ctcctscccc tcctsctcct
     6061 scccctcctg cccctcctgc tcctgcccct cctgcccctc ctgctcctgc ccctcctgct
     6121 cctgcccctc ctgctcctgc ccctcctgct cctscccctc ctscccctcc tscccctcct
     6181 cctgctcctg cccctcctgc tcctgcccct cctgcccctc ctgcccctcc tgctcctgcc
     6241 cctcctcctg ctcctgcccc tcctgcccct cctscccctc ctcctgctcc tgcccctcct
     6301 gcccctcctc ctgctcctgc ccctcctcct sctcctgccc ctcctscccc tcctscccct
     6361 cctcctsctc ctgcccctcc tscccctcct cctgctcctg cccctcctcc tgctcctgcc
     6421 cctcctgccc ctcctgcccc tcctcctgct cctgcccctc ctcctgctcc tgcccctcct
     6481 gcccctcctg cccctcctgc ccctcctcct gctcctgccc ctcctcctgc tcctgcccct
     6541 cctgctcctg cccctcccgc tcctgctcct gctcctgttc caccgtgggt ccctttgcag
     6601 ccaatgcaac ttggacgttt ttggggtctc cggacaccat ctctatgtct tggccctgat
     6661 cctgagccgc ccggggctcc tggtcttccg cctcctcgtc ctcgtcctct tccccgtcct
     6721 cgtccatggt tatcaccccc tcttctttga ggtccactgc cgccggagcc ttctggtcca
     6781 gatgtgtctc ccttctctcc taggccattt ccaggtcctg tacctggccc ctcgtcagac
     6841 atgattcaca ctaaaagaga tcaatagaca tctttattag acgacgctca gtgaatacag
     6901 ggagtgcaga ctcctgcccc ctccaacagc ccccccaccc tcatcccctt catggtcgct
     6961 gtcagacaga tccaggtctg aaaattcccc atcctccgaa ccatcctcgt cctcatcacc
     7021 aattactcgc agcccggaaa actcccgctg aacatcctca agatttgcgt cctgagcctc
     7081 aagccaggcc tcaaattcct cgtccccctt tttgctggac ggtagggatg gggattctcg
     7141 ggacccctcc tcttcctctt caaggtcacc agacagagat gctactgggg caacggaaga
     7201 aaagctgggt gcggcctgtg aggatcagct tatcgatgat aagctgtcaa acatgagaat
     7261 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     7321 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     7381 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     7441 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     7501 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     7561 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     7621 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     7681 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     7741 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     7801 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     7861 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     7921 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     7981 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     8041 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     8101 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     8161 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     8221 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     8281 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     8341 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     8401 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     8461 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     8521 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     8581 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     8641 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     8701 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     8761 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     8821 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     8881 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     8941 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     9001 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     9061 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acg
//
