LOCUS       40924_8386        6544 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCAGGS-FLAGmA20f5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6544)
  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  JOURNAL   Unpublished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 6544)
  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 6544)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6544)
  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            "Unpublished"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..6544
                     /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..1815
                     /codon_start=1
                     /note="unnamed protein product; mA20 (1-590)"
                     /protein_id="SJL88014.1"
                     /translation="MAEQLLPQALYLSNMRKAVKIRERTPEDIFKPTNGIIYHFKTMHR
                     YTLEMFRTCQFCPQFREIIHKALIDRSVQASLESQKKLNWCREVRKLVALKTNGDGNCL
                     MHAACQYMWGVQDTDLVLRKALCSTLKETDTRNFKFRWQLESLKSQEFVETGLCYDTRN
                     WNDEWDNLVKMASADTPAARSGLQYNSLEEIHIFVLSNILRRPIIVISDKMLRSLESGS
                     NFAPLKVGGIYLPLHWPAQECYRYPIVLGYDSQHFVPLVTLKDSGPELRAVPLVNRDRG
                     RFEDLKVHFLTDPENEMKEKLLKEYLIVMEIPVQGWDHGTTHLINAAKLDEANLPKEIN
                     LVDDYFELVQHEYKKWQENSDQARRAAHAQNPLEPSTPQLSLMDIKCETPNCPFFMSVN
                     TQPLCHECSERRQKNQSKLPKLNSKLGPEGLPGVGLGSSNWSPEETAGGPHSAPPTAPS
                     LFLFSETTAMKCRSPGCPFTLNVQHNGFCERCHARQINASHTADPGKCQACLQDVTRTF
                     NGICSTCFKRTTAEPSSSLTSSIPASCHQRSKSDPSQLIQSLTPHSCHRTGNVSPSGCL
                     SQAARTPGDRAGTS"
     polyA_signal    1897..1952
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2313..2329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2337..2353)
                     /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(2361..2391)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2405..2426)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2484..2680
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    2686..2820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3058..3646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3820..4677)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4678..4782)
                     /label=AmpR promoter
     enhancer        4817..5196
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      4817..5194
                     /label=hCMV-IE enhancer
                     /note="hCMV-IE enhancer"
                     /regulatory_class="promoter"
     promoter        5198..5475
                     /label=chicken beta-actin promoter
     intron          5476..6493
                     /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 caacgctcca agtctgaccc ctcacaactc atccaaagtc tcactccaca ctcttgccac
     1741 cggactggaa atgtctctcc ttctggctgc ctctcccagg ctgcacggac tccaggagac
     1801 agagcaggga caagctaagt gtcgacgaag atctttttcc ctctgccaaa aattatgggg
     1861 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     1921 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     1981 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     2041 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     2101 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     2161 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     2221 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     2281 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2341 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctgggtg
     2401 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2461 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     2521 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2581 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     2641 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     2701 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2761 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     2821 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2881 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2941 ctcactcaaa ggcggtaata cggttatcca cagaatcagg gataacgcag gaaagaacat
     3001 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3481 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3841 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3901 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3961 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4021 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4081 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4141 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4201 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4261 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4321 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4381 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4441 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4501 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4561 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4621 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4681 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4741 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4801 gccacctggg tcgatcgaca ttgattattg actagttatt aatagtaatc aattacgggg
     4861 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     4921 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     4981 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     5041 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     5101 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     5161 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     5221 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     5281 ttattttgtg cagcgatggg ggcggggggg gggggggggc cccccccagg cggggcgggg
     5341 cggggcgagg ggcggggcgg ggcgaggcgg aaaggtgcgg cggcagccaa tcagagcggc
     5401 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
     5461 agcgcgcggc gggcgggagt cgttgcgcgc tgccttcccc ccgtgccccg ctccgccgcc
     5521 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
     5581 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
     5641 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
     5701 tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc
     5761 ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag
     5821 gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc
     5881 tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg
     5941 caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg
     6001 ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg
     6061 gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaagg ggcgcggcgg
     6121 cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta
     6181 atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg
     6241 gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa
     6301 ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca
     6361 gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
     6421 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt
     6481 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa
     6541 agaa
//
