LOCUS       40924_8366        6640 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGS-FLAGmA20d5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  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  JOURNAL   Unpublished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 6640)
  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 6640)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6640)
  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            "Unpublished"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..6640
                     /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..1914
                     /codon_start=1
                     /note="unnamed protein product; mA20 (d547-699)"
                     /protein_id="SJL87289.1"
                     /translation="MAEQLLPQALYLSNMRKAVKIRERTPEDIFKPTNGIIYHFKTMHR
                     YTLEMFRTCQFCPQFREIIHKALIDRSVQASLESQKKLNWCREVRKLVALKTNGDGNCL
                     MHAACQYMWGVQDTDLVLRKALCSTLKETDTRNFKFRWQLESLKSQEFVETGLCYDTRN
                     WNDEWDNLVKMASADTPAARSGLQYNSLEEIHIFVLSNILRRPIIVISDKMLRSLESGS
                     NFAPLKVGGIYLPLHWPAQECYRYPIVLGYDSQHFVPLVTLKDSGPELRAVPLVNRDRG
                     RFEDLKVHFLTDPENEMKEKLLKEYLIVMEIPVQGWDHGTTHLINAAKLDEANLPKEIN
                     LVDDYFELVQHEYKKWQENSDQARRAAHAQNPLEPSTPQLSLMDIKCETPNCPFFMSVN
                     TQPLCHECSERRQKNQSKLPKLNSKLGPEGLPGVGLGSSNWSPEETAGGPHSAPPTAPS
                     LFLFSETTAMKCRSPGCPFTLNVQHNGFCERCHARQINASHTADPGKCQACLQDVTRTF
                     NGICSTCFKRTTAEPSSSLTSSIPASCHQKCARASCKNILACRSEELCMECQHLSQRVG
                     SVAHRGEPTPEEPPKQRCRAPACDHFGNAKCNGYCNECYQFKQMYG"
     polyA_signal    1993..2048
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2409..2425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2433..2449)
                     /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(2457..2487)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2501..2522)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2580..2776
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    2782..2916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3154..3742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3916..4773)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4774..4878)
                     /label=AmpR promoter
     enhancer        4913..5292
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      4913..5290
                     /label=hCMV-IE enhancer
                     /note="hCMV-IE enhancer"
                     /regulatory_class="promoter"
     promoter        5294..5571
                     /label=chicken beta-actin promoter
     intron          5572..6589
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
ORIGIN
        1 ttcctcgaga tggactacaa ggacgacgat gacaagggta ccgccatggc tgaacaactt
       61 cttcctcagg ctttgtattt gagcaatatg cggaaagctg tgaagatacg agagagaacc
      121 ccagaagaca ttttcaaacc taccaatggg atcatctatc actttaaaac catgcaccga
      181 tacacgctgg agatgttcag aacatgccag ttttgcccac agttccgaga gatcatccac
      241 aaagcactta ttgacagaag tgtccaggct tccctggaaa gccagaagaa gctcaactgg
      301 tgtcgtgaag tcaggaagct cgtggctctg aaaaccaatg gtgatggaaa ctgcctcatg
      361 catgcagctt gtcagtacat gtggggtgtt caggatactg acctggtcct gaggaaggcc
      421 ctctgcagca cccttaagga gacagacact cggaacttta aattccgctg gcagctggaa
      481 tctctgaaat ctcaggaatt tgtggaaaca ggactttgct acgacactcg gaactggaat
      541 gacgaatggg acaacttggt caaaatggca tcagcagaca cacctgcagc ccgaagtgga
      601 cttcagtaca attccctgga agaaatccac atatttgtcc tcagcaacat cctcagaaga
      661 cccatcattg tcatttcaga caaaatgcta agaagtttgg aatctggttc caattttgct
      721 cctttgaaag tgggtgggat ttatctgcct cttcactggc ctgcccagga gtgttacaga
      781 tatcccatcg tcctaggcta tgacagccag cactttgtac ccctggtgac cctgaaggac
      841 agtggacctg aacttcgcgc tgttccactt gttaacagag accggggtag gtttgaagac
      901 ttaaaagttc acttcttgac agatcctgag aatgagatga aggaaaagct tctaaaggag
      961 tacttgatag tgatggagat ccctgtgcaa ggctgggacc acggcacgac tcacctgatc
     1021 aacgctgcaa aattggatga agctaactta cccaaagaaa taaatttggt agacgattac
     1081 tttgagcttg ttcagcacga atacaagaaa tggcaggaga acagcgatca ggccaggaga
     1141 gcggcacatg cgcagaaccc cttggagcct tccacacccc agctatcact catggatata
     1201 aaatgtgaga cacccaactg tcctttcttc atgtccgtga acactcagcc tttatgccac
     1261 gaatgctcag agaggcgcca aaagaatcag agcaagctcc caaagctgaa ctcgaagcta
     1321 ggccctgaag gactcccagg cgtgggactt ggctcctcaa actggagccc cgaggaaacc
     1381 gctggaggac ctcattcagc cccacccaca gcacccagcc tttttctctt cagtgagacc
     1441 actgcaatga agtgcaggag tcctgggtgc ccttttactt tgaatgtgca gcataatgga
     1501 ttctgtgagc gttgccacgc ccggcagatt aatgccagcc acaccgcaga ccctggaaag
     1561 tgccaagcct gccttcagga tgtcactcgg acctttaatg gcatctgcag tacctgtttc
     1621 aaaaggacta cagcagagcc cagctccagc ctcacttcca gtatccctgc ctcctgtcac
     1681 caaaaatgtg cccgggcctc ctgcaagaac attctggcct gtcgcagtga ggaactctgt
     1741 atggagtgcc agcacctaag ccaacgagta ggttctgtgg cccaccgggg tgagcccacg
     1801 cctgaagagc cccctaaaca gcgctgccgg gcccctgctt gtgatcactt tggcaatgcc
     1861 aagtgtaatg gttactgcaa tgagtgctac cagttcaagc agatgtatgg ctaagtgtcg
     1921 acgaagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     1981 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     2041 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     2101 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     2161 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     2221 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     2281 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     2341 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     2401 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2461 atacgagccg gaagcataaa gtgtaaagcc tgggtgccta atgagtgagc taactcacat
     2521 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc
     2581 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2641 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2701 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2761 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
     2821 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2881 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
     2941 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3001 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3061 tatccacaga atcagggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga tcgacattga
     4921 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     4981 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     5041 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     5101 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     5161 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     5221 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     5281 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     5341 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     5401 gggggggggg gggggccccc cccaggcggg gcggggcggg gcgaggggcg gggcggggcg
     5461 aggcggaaag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     5521 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgtt
     5581 gcgcgctgcc ttccccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
     5641 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     5701 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
     5761 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
     5821 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     5881 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
     5941 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
     6001 gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc
     6061 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     6121 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     6181 cgggccgggg agggctcggg ggaaggggcg cggcggcccc cggagcgccg gcggctgtcg
     6241 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact
     6301 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag
     6361 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg
     6421 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga
     6481 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     6541 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     6601 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
//
