LOCUS       40924_13285        5611 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCR3Flag-E8-FLIP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5611)
  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  JOURNAL   Unpublished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 5611)
  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 5611)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5611)
  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            "Unpublished"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..5611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             705..728
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             735..1250
                     /codon_start=1
                     /note="unnamed protein product; E8-FLIP"
                     /protein_id="SJL88342.1"
                     /translation="MSHYSMIDTYFSLDEDETETYLYLCRDLLKNKGEFQCTRDAFKFL
                     SDYACLSAANQMELLFRVGRLDLIRRIFGQTWTPDSCPRYYMPICSPFRCLMALVNDFL
                     SDKEVEEMYFLCAPRLESHLEPGSKKSFLRLASLLEDLELLGGDKLTFLRHLLTTIGRA
                     DLVKNLQV"
     promoter        complement(1326..1344)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    1370..1594
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1726..2314)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(2643..2690)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(2925..3710)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(3745..4102)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             complement(4165..5022)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5023..5127)
                     /label=AmpR promoter
     rep_origin      5154..5609
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg gtaccgagct cggatccccg ggaatcaaaa catggattac aaagacgatg
      721 acgataaaga attcatgagt cactacagta tgattgacac ttacttctcc ctggacgagg
      781 atgagaccga gacctatctg tacctctgca gggacctcct gaaaaataag ggagagttcc
      841 agtgcaccag ggacgccttt aaatttctgt cagactatgc gtgcctctcg gccgccaacc
      901 agatggagct attgtttagg gtggggaggc tggacttgat aaggagaatc tttggccaga
      961 cctggacgcc agactcgtgc cccaggtact acatgcccat ctgctccccc ttcaggtgcc
     1021 tgatggccct ggtcaatgac ttcctctcgg acaaggaggt ggaggagatg tactttctgt
     1081 gcgccccgcg cctggagagt cacttagagc cgggctccaa aaagtctttc ctcagactgg
     1141 ccagtctgct ggaagacctg gagctgctgg ggggcgataa gctaacattt ttaaggcacc
     1201 tcttgaccac catcggacgg gccgacctgg taaaaaatct acaggtttag ggatccgaat
     1261 aagccgaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc
     1321 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     1381 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1441 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1501 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1561 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagtggc
     1621 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1681 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1741 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1801 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1861 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1921 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1981 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2041 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2101 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2161 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2221 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2281 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2341 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2401 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2461 atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg
     2521 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc
     2581 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
     2641 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg
     2701 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa
     2761 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     2821 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     2881 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     2941 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     3001 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     3061 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     3121 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     3181 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttga
     3241 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     3301 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     3361 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     3421 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     3481 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     3541 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     3601 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     3661 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     3721 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     3781 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     3841 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     3901 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     3961 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     4021 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     4081 cagctggttc tttccgcctc aggactcttc ctttttcaat aaatcaatct aaagtatata
     4141 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4201 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4261 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4321 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4381 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4441 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4501 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4561 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4621 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4681 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4741 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4801 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4861 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4921 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4981 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5041 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5101 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     5161 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5221 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5281 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5341 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5401 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5461 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5521 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5581 ttttaacaaa atattaacgc ttacaattta c
//
