LOCUS       40924_13255        7412 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian vector pCR3-GW-GST-C, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7412)
  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  JOURNAL   Unpublished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 7412)
  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 7412)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7412)
  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            "Unpublished"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..7412
                     /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"
     protein_bind    722..846
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        871..901
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             955..1611
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1956..2258
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2302..2426)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2458..3108
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     promoter        complement(3127..3145)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3171..3395
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3527..4115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(4444..4491)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(4726..5511)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDQEHQGL
                     APAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALA
                     TRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5546..5903)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             complement(5966..6823)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6824..6928)
                     /label=AmpR promoter
     rep_origin      6955..7410
                     /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 cggatccact agtccagtgt ggtggaattc tgcagatatc
      721 aacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt
      781 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
      841 actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
      901 gtgtggattt tgagttagga tccggcgaga ttttcaggag ctaaggaagc taaaatggag
      961 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     1021 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     1081 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1141 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     1201 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1261 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1321 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1381 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1441 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1501 gtgctgatgc cgctggcgat tcaggttcat catgccgtct gtgatggctt ccatgtcggc
     1561 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaagatct
     1621 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     1681 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtgt gctatgaagc
     1741 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     1801 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     1861 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     1921 cggctctttt gctgacgaga acagggactg gtgaaatgca gtttaaggtt tacacctata
     1981 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     2041 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     2101 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     2161 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     2221 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tccgttatac
     2281 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     2341 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     2401 cgttcagctt tcttgtacaa agtggttgat atccagcaca gtggcggccg ctcgagctcc
     2461 cctatactag gttattggaa aattaagggc cttgtgcaac ccactcgact tcttttggaa
     2521 tatcttgaag aaaaatatga agagcatttg tatgagcgcg atgaaggtga taaatggcga
     2581 aacaaaaagt ttgaattggg tttggagttt cccaatcttc cttattatat tgatggtgat
     2641 gttaaattaa cacagtctat ggccatcata cgttatatag ctgacaagca caacatgttg
     2701 ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg aaggagcggt tttggatatt
     2761 agatacggtg tttcgagaat tgcatatagt aaagactttg aaactctcaa agttgatttt
     2821 cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca taaaacatat
     2881 ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt atgacgctct tgatgttgtt
     2941 ttatacatgg acccaatgtg cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt
     3001 attgaagcta tcccacaaat tgataagtac ttgaaatcca gcaagtatat agcatggcct
     3061 ttgcagggct ggcaagccac gtttggtggt ggcgaccatc ctccaaaatg atctagaggg
     3121 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     3181 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3241 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3301 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3361 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagtgg
     3421 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3481 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3541 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3601 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3661 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3721 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3781 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3841 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3901 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3961 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4021 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4081 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4141 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4201 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4261 tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg
     4321 ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc
     4381 cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt
     4441 tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc
     4501 gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga
     4561 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
     4621 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
     4681 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
     4741 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     4801 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     4861 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     4921 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     4981 gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttg
     5041 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     5101 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     5161 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     5221 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     5281 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     5341 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     5401 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     5461 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     5521 tcctcatcct gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc
     5581 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
     5641 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
     5701 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
     5761 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
     5821 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     5881 acagctggtt ctttccgcct caggactctt cctttttcaa taaatcaatc taaagtatat
     5941 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6001 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6061 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6121 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6181 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6241 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6301 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6361 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6421 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6481 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6541 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6601 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6661 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6721 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6781 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6841 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6901 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     6961 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     7021 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     7081 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     7141 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     7201 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     7261 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     7321 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     7381 attttaacaa aatattaacg cttacaattt ac
//
