LOCUS       40924_10501        6448 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian vector pCFP-N-GW, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6448)
  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  JOURNAL   Unpublished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 6448)
  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 6448)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6448)
  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            "Unpublished"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..6448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      39..365
                     /label=pMB1 ori
                     /note="pMB1 ori"
     enhancer        432..735
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        736..939
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             984..1700
                     /codon_start=1
                     /label=ECFP
                     /note="enhanced CFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    1719..1843
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1868..1898
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1952..2608
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2953..3255
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3299..3423)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    3427..3483
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3607..3728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3735..4190)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4217..4321
                     /label=AmpR promoter
     promoter        4323..4680
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4715..5506
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5741..5788
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6117..6448
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
       61 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      121 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      181 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      241 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      301 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
      361 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
      421 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      481 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      541 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      601 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      661 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      721 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      781 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      841 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      901 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      961 cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1021 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1081 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1141 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca
     1201 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1261 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1321 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1381 aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata
     1441 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1501 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1561 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1621 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1681 gcatggacga gctgtacaag tccggactca gatcatcaac aagtttgtac aaaaaagctg
     1741 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
     1801 agactacata atactgtaaa acacaacata tccagtcact atggcggccg cattaggcac
     1861 cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga gttaggatcc
     1921 ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
     1981 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     2041 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     2101 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
     2161 tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
     2221 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
     2281 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     2341 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg
     2401 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     2461 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
     2521 ggttcatcat gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
     2581 gtactgcgat gagtggcagg gcggggcgta aagatctgga tccggcttac taaaagccag
     2641 ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata ctgatatgta
     2701 tacccgaagt atgtcaaaaa gaggtgtgct atgaagcagc gtattacagt gacagttgac
     2761 agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt ctggtaagca
     2821 caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg
     2881 aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct gacgagaaca
     2941 gggactggtg aaatgcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg
     3001 tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt gatccccctg
     3061 gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc
     3121 ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc
     3181 ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg
     3241 atgttctggg gaatataaat gtcaggctcc gttatacaca gccagtctgc aggtcgacca
     3301 tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta
     3361 atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct tgtacaaagt
     3421 ggttgatgat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga
     3481 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     3541 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3601 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3661 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3721 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3781 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3841 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3901 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3961 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4021 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4081 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4141 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4201 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4261 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4321 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4441 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4501 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4561 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4621 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4681 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4741 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4801 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4861 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4921 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4981 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5041 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5101 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5161 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5221 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5281 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5341 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5401 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5461 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5521 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5581 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5641 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5701 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5761 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5821 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5881 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5941 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6001 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6061 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6121 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6181 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6241 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     6301 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6361 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6421 cagcggtcgg gctgaacggg gggttcgt
//
