LOCUS       40924_37398        8807 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pRMCE-DV1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8807)
  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  JOURNAL   Unpublished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 8807)
  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 8807)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8807)
  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            "Unpublished"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..8807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    35..277
                     /label=chicken beta-globin 5'HS4 insulator core sequence
                     /note="chicken beta-globin 5'HS4 insulator core sequence"
     misc_feature    278..520
                     /label=chicken beta-globin 5'HS4 insulator core sequence
                     /note="chicken beta-globin 5'HS4 insulator core sequence"
     promoter        546..1045
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     regulatory      1059..1068
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1070..1117
                     /label=FRT mutant
                     /note="FRT mutant"
     primer_bind     complement(1162..1178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1186..1202)
                     /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(1210..1240)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1255..1276)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1564..2152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2326..3183)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3184..3288)
                     /label=AmpR promoter
     primer_bind     3762..3778
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3801..3848
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    3865..3968
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     primer_bind     4001..4017
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4031..4064
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4773..4907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(5005..5101)
                     /label=SV40 polyA late
                     /note="SV40 polyA late"
                     /regulatory_class="terminator"
     regulatory      5005..5101
                     /label=SV40 polyA early
                     /note="SV40 polyA early"
                     /regulatory_class="terminator"
     polyA_signal    5027..5161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      complement(5259..5355)
                     /label=SV40 polyA late
                     /note="SV40 polyA late"
                     /regulatory_class="terminator"
     regulatory      5259..5355
                     /label=SV40 polyA early
                     /note="SV40 polyA early"
                     /regulatory_class="terminator"
     polyA_signal    5281..5415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(5444..5477)
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(5444..5477)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(5512..5528)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5533..5657
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5682..5712
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             5766..6422
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             6767..7069
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(7113..7237)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     7242..7258
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7281..7775
                     /label=5' UTR of the EMCV polyprotein gene (IRES)
                     /note="5' UTR of the EMCV polyprotein gene (IRES)"
     CDS             7788..8504
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    8538..8762
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgcgccgggc cctcgcgact cgaggaataa attcgagctc acggggacag ccccccccca
       61 aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg
      121 ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc
      181 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
      241 agcctgcaga cacctggggg atacggggaa aaagcttgag ctcacgggga cagccccccc
      301 ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc
      361 cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac
      421 agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct
      481 ttgagcctgc agacacctgg gggatacggg gaaaaagctt gtcgacgaat aaattcaatt
      541 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
      601 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
      661 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
      721 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
      781 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
      841 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
      901 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
      961 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
     1021 ctcttcctca tctccgggcc tttcgacctg caatcgccgc caccatggag aagttcctat
     1081 tccgaagttc ctattcttca aaaggtatag gaacttccga tcgggatccc caatcctgca
     1141 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1201 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1261 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1321 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1381 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1801 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1981 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3061 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3301 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3361 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     3421 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     3481 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     3541 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     3601 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3661 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3721 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3781 attccacaaa agtgcgatcg gaagttccta ttccgaagtt cctattctct agaaagtata
     3841 ggaacttctt aattaaggga tctgtagggc gcagtagtcc agggtttcct tgatgatgtc
     3901 atacttatcc tgtccctttt ttttccacag ctcgcggttg aggacaaact cttcgcggtc
     3961 tttccagtgg ggatcgacgg tatcgtagag tcgaggccgc tctagaacta gtggatccgg
     4021 aacccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtc cctcgacctg
     4081 caggaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta
     4141 gcagccccgc tggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
     4201 ccaccggtag cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc
     4261 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat
     4321 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg
     4381 ggtaggatca taatttaaac aagcaaaacc aaattaaggg ccagctcatt cctcccactc
     4441 atgatctata gatctataga tctctcgtgg gatcattgtt tttctcttga ttcccacttt
     4501 gtggttctaa gtactgtggt ttccaaatgt gtcagtttca tagcctgaag aacgagatca
     4561 gcagcctctg ttccacatac acttcattct cagtattgtt ttgccaagtt ctaattccat
     4621 cagaagcttg cagatctgcg actctagagg atctgcgact ctagaggatc ataatcagcc
     4681 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4741 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4801 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4861 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctgc gactctagag
     4921 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     4981 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     5041 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5101 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     5161 ctgcgactct agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
     5221 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     5281 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5341 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5401 tatcatgtct ggatccccat caagctgatc cggaaccctt aatataactt cgtataatgt
     5461 atgctatacg aagttattag gtccctcgac ctgcagccca agctagctta tcgataccgt
     5521 cgacctcgaa tcacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     5581 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     5641 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     5701 gctcgtataa tgtgtggatt ttgagttagg atccggcgag attttcagga gctaaggaag
     5761 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     5821 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     5881 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     5941 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     6001 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     6061 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     6121 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     6181 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     6241 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     6301 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc tgtgatggct
     6361 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     6421 cgtaaacgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     6481 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggtg
     6541 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     6601 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     6661 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     6721 attgaaatga acggctcttt tgctgacgag aacagggact ggtgaaatgc agtttaaggt
     6781 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     6841 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     6901 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     6961 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     7021 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     7081 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     7141 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     7201 tttacgtttc tcgttcagct ttcttgtaca aagtggtgat tcgaggtcga cggtatcgat
     7261 aagcttgata tcgaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     7321 cgcttggaat aaggccggtg agcgtttgtc tatatgttat tttccaccat attgccgtct
     7381 tttggcaatg tgagggcccg aaacctggcc ctgtcttctt gacgacgatt cctaggggtc
     7441 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     7501 tggaagcttc ttgaagacaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     7561 acctggcgac atggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     7621 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     7681 gcacatgctt tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg
     7741 ggacgtggtt ttggtttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg
     7801 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     7861 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     7921 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     7981 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     8041 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     8101 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     8161 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     8221 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     8281 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     8341 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     8401 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     8461 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgagc
     8521 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     8581 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     8641 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     8701 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     8761 ggcttctgag gcggaaagaa ccagctgggg ctcgatcctc tagttgg
//
