LOCUS       62056_116        5355 bp DNA     circular SYN 26-AUG-2019
DEFINITION  Cloning vector 35SpGW, complete sequence.
ACCESSION   MK450604
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5355)
  AUTHORS   Siqueira Reis R.
  TITLE     CMV 35S promoter-driven Gateway(R) plasmid for protoplast 
            transformation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5355)
  AUTHORS   Siqueira Reis R.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-JAN-2019) Department of Plant Molecular Biology, 
            University of Lausanne, Quartier UNIL-Sorge, Batiment Biophore, 
            Lausanne, Vaud 1015, Switzerland
REFERENCE   3  (bases 1 to 5355)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-JAN-2019) Department of Plant Molecular Biology, University of 
            Lausanne, Quartier UNIL-Sorge, Batiment Biophore, Lausanne, Vaud 
            1015, Switzerland"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    358..457
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     550..566
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        666..1337
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     protein_bind    1397..1521
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1546..1576
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1630..2286
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2631..2933
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2977..3101)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3167..3414
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    complement(3463..3562)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3684..3754
                     /label=AmpR promoter
     CDS             3755..4612
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4705..5293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctggc gccggaacca attcagtcga
      481 ctggatccgg taccgaattc gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      541 cacgacgttg taaaacgacg gccagtgcca agcttggcgt gcctgcaggt caacatggtg
      601 gagcacgaca cacttgtcta ctccaaaaat atcaaagata cagtctcaga agaccaaagg
      661 gcaattgaga cttttcaaca aagggtaata tccggaaacc tcctcggatt ccattgccca
      721 gctatctgtc actttattgt gaagatagtg gaaaaggaag gtggctccta caaatgccat
      781 cattgcgata aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg tcccaaagat
      841 ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag
      901 caagtggatt gatgtgataa catggtggag cacgacacac ttgtctactc caaaaatatc
      961 aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag ggtaatatcc
     1021 ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa gatagtggaa
     1081 aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat cgttgaagat
     1141 gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
     1201 gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc cactgacgta
     1261 agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca
     1321 tttcatttgg agaggacctc gactctagag gatccccggg taccgggccc cccctcgagg
     1381 cgcgccaagc tatcaaacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat
     1441 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
     1501 acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct
     1561 tccggctcgt ataatgtgtg gattttgagt taggatccgg cgagattttc aggagctaag
     1621 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     1681 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     1741 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     1801 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
     1861 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     1921 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     1981 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     2041 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     2101 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     2161 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat
     2221 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     2281 ggggcgtaat ctagaggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc
     2341 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga
     2401 ggtgtgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca
     2461 aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg
     2521 tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg
     2581 gtttattgaa atgaacggct cttttgctga cgagaacagg gactggtgaa atgcagttta
     2641 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     2701 ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     2761 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     2821 tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca
     2881 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     2941 caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt
     3001 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
     3061 tcattttacg tttctcgttc agctttcttg tacaaagtgg ttcgataatt ccttaattaa
     3121 ctagttctag agcggccgcc caccgcggtg gagctcgaat ttccccgatc gttcaaacat
     3181 ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata
     3241 atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat
     3301 gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa
     3361 aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactgaattc
     3421 gtaatcatgg tcatagctgt ttcccgcact cgagatatct agacccagct ttcttgtaca
     3481 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     3541 gtcaaaataa aatcattatt tgccatccag ctgcagctct ggcccgtgtc tcaaaatctc
     3601 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     3661 ataaacagta atacaagggg tgttttcaaa tatgtatccg ctcatgagac aataaccctg
     3721 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3781 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3841 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3901 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3961 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     4021 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4081 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4141 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4201 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4261 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     4321 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4381 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4441 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4501 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4561 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaatcaga
     4621 attggttaat tggttgtaac attattcaga ttgggccccg ttccactgag cgtcagaccc
     4681 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4741 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4801 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4861 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4921 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4981 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5041 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5101 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5161 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5221 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5281 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5341 ttttgctcac atgtt
//
