LOCUS       40924_17059        6582 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pEF6A-E-PKR-ND, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6582)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6582)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 6582)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6582)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        351..1529
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        1546..1564
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1639..1677
                     /label=E-tag
                     /note="E-tag"
     CDS             1687..2442
                     /codon_start=1
                     /note="unnamed protein product; hPKR-ND"
                     /protein_id="SJL86758.1"
                     /translation="MAGDLSAGFFMEELNTYRQKQGVVLKYQELPNSGPPHDRRFTFQV
                     IIDGREFPEGEGRSKKEAKNAAAKLAVEILNKEKKAVSPLLLTTTNSSEGLSMGNYIGL
                     INRIAQKKRLTVNYEQCASGVHGPEGFHYKCKMGQKEYSIGTGSTKQEAKQLAAKLAYL
                     QILSEETSVKSDYLSSGSFATTCESQSNSLVTSTLASESSSEGDFSADTSEINSNSDSL
                     NSSSLLMNGLRNNQRKAKRSLAPRFDLPD"
     CDS             2464..2493
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             2509..2526
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    2555..2779
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2825..3253
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3267..3596
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        3644..3691
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3710..4105
                     /label=BSD
                     /note="blasticidin S deaminase"
     polyA_signal    4266..4399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4436..4452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4460..4476)
                     /label=lac operator
                     /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(4484..4514)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4529..4550)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4827..5415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5589..6446)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6447..6551)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac taggcttttg caaaaagctt tgcaaagatg gataaagttt taaacagaga
      301 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      361 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      421 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      481 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      541 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      601 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      661 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      721 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      781 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      841 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      901 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      961 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
     1021 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
     1081 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1141 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1201 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1261 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1321 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1381 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1441 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1501 gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata cgactcacta
     1561 tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac tagtccagtg
     1621 tggtggaatt ccaccatggg tgcgccggtg ccgtatccgg acccgctgga accgcgtgcg
     1681 gccgctatgg ctggtgatct ttcagcaggt ttcttcatgg aggaacttaa tacataccgt
     1741 cagaagcagg gagtagtact taaatatcaa gaactgccta attcaggacc tccacatgat
     1801 aggaggttta catttcaagt tataatagat ggaagagaat ttccagaagg tgaaggtaga
     1861 tcaaagaagg aagcaaaaaa tgccgcagcc aaattagctg ttgagatact taataaggaa
     1921 aagaaggcag ttagtccttt attattgaca acaacgaatt cttcagaagg attatccatg
     1981 gggaattaca taggccttat caatagaatt gcccagaaga aaagactaac tgtaaattat
     2041 gaacagtgtg catcgggggt gcatgggcca gaaggatttc attataaatg caaaatggga
     2101 cagaaagaat atagtattgg tacaggttct actaaacagg aagcaaaaca attggccgct
     2161 aaacttgcat atcttcagat attatcagaa gaaacctcag tgaaatctga ctacctgtcc
     2221 tctggttctt ttgctactac gtgtgagtcc caaagcaact ctttagtgac cagcacactc
     2281 gcttctgaat catcatctga aggtgacttc tcagcagata catcagagat aaattctaac
     2341 agtgacagtt taaacagttc ttcgttgctt atgaatggtc tcagaaataa tcaaaggaag
     2401 gcaaaaagat ctttggcacc cagatttgac cttcctgact aactgcagtc tagagggccc
     2461 ttcgaacaaa aactcatctc agaagaggat ctgaatatgc ataccggtca tcatcaccat
     2521 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2581 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2641 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2701 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2761 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2821 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2881 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2941 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3001 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3061 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3121 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3181 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3241 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3301 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3361 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3421 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3481 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3541 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3601 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     3661 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagcc
     3721 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     3781 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     3841 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     3901 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     3961 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     4021 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     4081 gccctctggt tatgtgtggg agggctaagc acttcgtggc cgaggagcag gactgacacg
     4141 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4201 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4261 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4321 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4381 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4441 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4501 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4561 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4621 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4681 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4741 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccaggaac
     4801 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4861 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4921 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4981 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5041 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5101 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5161 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5221 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5281 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5341 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5401 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5461 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5521 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5581 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5641 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5701 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5761 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5821 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5881 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5941 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6001 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6061 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6121 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6181 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6241 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6301 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6361 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6421 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6481 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6541 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
