LOCUS       40924_16974        6943 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pEF1hCASP-9S-His, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6943)
  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  JOURNAL   Unpublished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 6943)
  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 6943)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6943)
  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            "Unpublished"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..6943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        473..1651
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        1668..1686
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2574..2615
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             2625..2642
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    2671..2895
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2941..3369
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3383..3712
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3779..4570
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4749..4882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4919..4935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4943..4959)
                     /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(4967..4997)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5012..5033)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5310..5898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6072..6929)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(join(6930..6943,1..91))
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
      481 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
      541 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
      601 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
      661 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
      721 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
      781 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
      841 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
      901 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
      961 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     1021 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     1081 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     1141 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     1201 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     1261 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     1321 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     1381 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     1441 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     1501 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     1561 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     1621 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac
     1681 tatagggaga cccaagctgg ctaggtaagc ttggtaccga gctcggatcc actagtccag
     1741 tgtggtggaa ttccgccatg gacgaagcgg atcggcggct cctgcggcgg tgccggctgc
     1801 ggctggtgga agagctgcag gtggaccagc tctgggacgt cctgctgagc cgcgagctgt
     1861 tcaggcccca tatgatcgag gacatccagc gggcaggctc tggatctcgg cgggatcagg
     1921 ccaggcagct gatcatagat ctggagactc gagggagtca ggctcttcct ttgttcatct
     1981 cctgcttaga ggacacaggc caggacatgc tggcttcgtt tctgcgaact aacaggcaag
     2041 caggaaagtt gtcgaagcca accctagaaa accttacccc agtggtgctc agaccagaga
     2101 ttcgcaaacc agaggttctc agaccggaaa cacccagacc agtggacatt ggttctggag
     2161 gattcggtga tgtcgagcag aaagaccatg ggtttgaggt ggcctccact tcccctgaag
     2221 acgagtcccc tggcagtaac cccgagccag atgccacccc gttccaggaa ggtttgagga
     2281 ccttcgacca gctggacgcc atatctagtt tgcccacacc cagtgacatc tttgtgtcct
     2341 actctacttt cccaggtttt gtttcctgga gggaccccaa gagtggctcc tggtacgttg
     2401 agaccctgga cgacatcttt gagcagtggg ctcactctga agacctgcag tccctcctgc
     2461 ttagggtcgc taatgctgtt tcggtgaaag ggatttataa acagatgcct ggttgcttta
     2521 atttcctccg gaaaaaactt ttctttaaaa catcatctag agggcccttc gaaggtaagc
     2581 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     2641 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2701 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2761 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2821 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2881 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2941 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3001 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3061 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3121 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3181 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3241 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3301 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3361 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3421 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3541 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3601 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3661 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3721 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     3781 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3841 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3901 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3961 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4021 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4081 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4141 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4201 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4261 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4321 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4381 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4441 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4501 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4561 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     4621 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4681 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4741 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4801 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4861 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4921 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4981 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5041 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5101 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5161 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5221 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagg
     5281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5941 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6001 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6061 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6121 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6181 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6241 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6301 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6361 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6421 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6481 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6541 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6601 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6661 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6721 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6781 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6841 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6901 agggcgacac ggaaatgttg aatactcata ctcttccttt ttc
//
