LOCUS       40924_8311        5000 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGS-FLAG-hUb-K27, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5000)
  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  JOURNAL   Unpublished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 5000)
  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 5000)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5000)
  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            "Unpublished"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..5000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             13..36
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             46..273
                     /label=ubiquitin-K27
                     /note="human ubiquitin variant containing Lys-27, with all
                     other lysines mutated to arginine"
     polyA_signal    353..408
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(769..785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(793..809)
                     /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(817..847)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(861..882)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        940..1136
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    1142..1276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1514..2102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2276..3133)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3134..3238)
                     /label=AmpR promoter
     enhancer        3273..3652
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      3273..3650
                     /label=hCMV-IE enhancer
                     /note="hCMV-IE enhancer"
                     /regulatory_class="promoter"
     promoter        3654..3931
                     /label=chicken beta-actin promoter
     intron          3932..4949
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
ORIGIN
        1 ttcctcgaga tggactacaa ggacgacgat gacaagggta ccgcgatgca gatcttcgtg
       61 aggaccctga ccggcaggac catcactctg gaggtggagc ccagtgacac catcgaaaat
      121 gtgaaggcca gaatccaaga tagggaaggc atcccccccg accagcagag gctcatcttt
      181 gcaggcaggc agctggaaga tggccgcact ctttctgact acaacatcca gagggagtcg
      241 accctgcacc tggtcctgcg tctcagaggt gggtaggtcg acgaagatct ttttccctct
      301 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
      361 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
      421 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
      481 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
      541 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
      601 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
      661 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
      721 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
      781 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
      841 gtgtaaagcc tgggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      901 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
      961 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1021 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     1081 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     1141 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     1201 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     1261 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1321 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1381 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcagggata
     1441 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1501 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1561 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1621 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1681 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1741 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1801 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1861 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1921 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1981 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2041 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2101 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2161 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2221 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2281 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2341 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2401 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2461 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2521 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2581 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2641 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2701 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2761 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2821 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2881 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2941 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3001 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3061 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3121 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3181 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3241 catttccccg aaaagtgcca cctgggtcga tcgacattga ttattgacta gttattaata
     3301 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     3361 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     3421 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     3481 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     3541 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     3601 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     3661 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     3721 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggccccc
     3781 cccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggaaag gtgcggcggc
     3841 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     3901 gccctataaa aagcgaagcg cgcggcgggc gggagtcgtt gcgcgctgcc ttccccccgt
     3961 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
     4021 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg
     4081 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt
     4141 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg
     4201 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct
     4261 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg
     4321 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg
     4381 cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc
     4441 ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg
     4501 gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg agggctcggg
     4561 ggaaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc
     4621 attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg
     4681 cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt
     4741 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
     4801 cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac
     4861 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
     4921 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     4981 ctcatcattt tggcaaagaa
//
