LOCUS       40924_26511        8981 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pK7WG_ASX, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8981)
  AUTHORS   Lukan T, Machens F, Coll A, Baebler S, Messerschmidt K, Gruden K.
  TITLE     Plant X-tender: An extension of the AssemblX system for the assembly
            and expression of multigene constructs in plants
  JOURNAL   PLoS ONE 13 (1), e0190526 (2018)
  PUBMED    29300787
REFERENCE   2  (bases 1 to 8981)
  AUTHORS   Lukan T, Machens F, Coll A, Baebler S, Messerschmidt K, Gruden K.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-NOV-2017) Department of Biotechnology and Systems 
            Biology, National Institute of Biology, Vecna pot 111, Ljubljana 
            1000, Slovenia
REFERENCE   3  (bases 1 to 8981)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8981)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2018"; volume: "13"; issue: "1"; pages: "e0190526"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-NOV-2017) Department of Biotechnology and Systems Biology, 
            National Institute of Biology, Vecna pot 111, Ljubljana 1000, 
            Slovenia"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1331..1957
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2394..3458
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      3527..3721
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    4065..4205
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4391..4979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5226..6014)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6542..6566
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(6659..6911)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(6954..7745)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(7779..7958)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    8067..8084
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     misc_feature    8103..8152
                     /label=homology region A0
                     /note="homology region A0"
     misc_feature    8153..8158
                     /label=HindIII restriction site
                     /note="HindIII restriction site"
     CDS             8488..8790
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    8822..8827
                     /label=HindIII restriction site
                     /note="HindIII restriction site"
     misc_feature    8828..8877
                     /label=homology region B0
                     /note="homology region B0"
     misc_feature    8878..8895
                     /label=I-SceI restriction site
                     /note="I-SceI restriction site"
     misc_feature    8958..8981
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 cctaagagaa aagagcgttt attagaataa cggatattta aaagggcgtg aaaaggttta
       61 tccgttcgtc catttgtatg tgcatgccaa ccacagggtt cccctcggga tcaaagtact
      121 ttaaagtact ttaaagtact ttaaagtact ttgatccaac ccctccgctg ctatagtgca
      181 gtcggcttct gacgttcagt gcagccgtct tctgaaaacg acatgtcgca caagtcctaa
      241 gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt
      301 cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc
      361 gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa
      421 cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc
      481 aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg
      541 acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag
      601 cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc
      661 acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc
      721 ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt
      781 ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag
      841 gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac
      901 cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc
      961 cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag
     1021 gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga
     1081 tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac gtctcaaccg
     1141 tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca
     1201 gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa
     1261 acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca
     1321 aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac
     1381 catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga
     1441 ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac
     1501 cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga
     1561 cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc
     1621 agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga
     1681 cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt
     1741 tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc
     1801 cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac
     1861 tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt
     1921 ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa
     1981 atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct
     2041 gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg
     2101 gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag
     2161 ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag
     2221 atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc
     2281 cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgt
     2341 ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt gacggtcgca
     2401 aaccatccgg cccggtacaa atcggcgcgg cgctgggtga tgacctggtg gagaagttga
     2461 aggccgcgca ggccgcccag cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt
     2521 ggcaagcggc cgctgatcga atccgcaaag aatcccggca accgccggca gccggtgcgc
     2581 cgtcgattag gaagccgccc aagggcgacg agcaaccaga ttttttcgtt ccgatgctct
     2641 atgacgtggg cacccgcgat agtcgcagca tcatggacgt ggccgttttc cgtctgtcga
     2701 agcgtgaccg acgagctggc gaggtgatcc gctacgagct tccagacggg cacgtagagg
     2761 tttccgcagg gccggccggc atggccagtg tgtgggatta cgacctggta ctgatggcgg
     2821 tttcccatct aaccgaatcc atgaaccgat accgggaagg gaagggagac aagcccggcc
     2881 gcgtgttccg tccacacgtt gcggacgtac tcaagttctg ccggcgagcc gatggcggaa
     2941 agcagaaaga cgacctggta gaaacctgca ttcggttaaa caccacgcac gttgccatgc
     3001 agcgtacgaa gaaggccaag aacggccgcc tggtgacggt atccgagggt gaagccttga
     3061 ttagccgcta caagatcgta aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc
     3121 tagctgattg gatgtaccgc gagatcacag aaggcaagaa cccggacgtg ctgacggttc
     3181 accccgatta ctttttgatc gatcccggca tcggccgttt tctctaccgc ctggcacgcc
     3241 gcgccgcagg caaggcagaa gccagatggt tgttcaagac gatctacgaa cgcagtggca
     3301 gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc
     3361 tgccggagta cgatttgaag gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct
     3421 accgcaacct gatcgagggc gaagcatccg ccggttccta atgtacggag cagatgctag
     3481 ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggtct ctttcctgtg gatagcacgt
     3541 acattgggaa cccaaagccg tacattggga accggaaccc gtacattggg aacccaaagc
     3601 cgtacattgg gaaccggtca cacatgtaag tgactgatat aaaagagaaa aaaggcgatt
     3661 tttccgccta aaactcttta aaacttatta aaactcttaa aacccgcctg gcctgtgcat
     3721 aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc
     3781 gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg
     3841 ctggcctacg gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc
     3901 gccggcgccc acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3961 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4021 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     4081 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     4141 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4201 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4261 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4321 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4381 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4441 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4501 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4561 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4621 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4681 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4741 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4801 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4861 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4921 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4981 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5041 ggattttggt catgcatgat atatctccca atttgtgtag ggcttattat gcacgcttaa
     5101 aaataataaa agcagacttg acctgatagt ttggctgtga gcaattatgt gcttagtgca
     5161 tctaatcgct tgagttaacg ccggcgaagc ggcgtcggct tgaacgaatt tctagctaga
     5221 cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac
     5281 tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca
     5341 agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc
     5401 ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca
     5461 tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc
     5521 gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc
     5581 aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg
     5641 gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg
     5701 cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca
     5761 gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa
     5821 gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg
     5881 tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt
     5941 tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc
     6001 accgcttccc ccatgatgtt taactttgtt ttagggcgac tgccctgctg cgtaacatcg
     6061 ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc
     6121 cgaggcatag actgtacccc aaaaaaacat gtcataacaa gaagccatga aaaccgccac
     6181 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg acggcagtta
     6241 cgctacttgc attacagctt acgaaccgaa cgaggcttat gtccactggg ttcgtgcccg
     6301 aattgatcac aggcagcaac gctctgtcat cgttacaatc aacatgctac cctccgcgag
     6361 atcatccgtg tttcaaaccc ggcagcttag ttgccgttct tccgaatagc atcggtaaca
     6421 tgagcaaagt ctgccgcctt acaacggctc tcccgctgac gccgtcccgg actgatgggc
     6481 tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg
     6541 gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc
     6601 ggacgttttt aatgtactga attaacgccg aattgaatta tcagcttgca tgccggtcga
     6661 tctagtaaca tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg
     6721 ttttctatcg cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata
     6781 aataacgtca tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat
     6841 atgataatca tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg
     6901 tttgaacgat ctgcttgact ctagctagag tccgaacccc agagtcccgc tcagaagaac
     6961 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     7021 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     7081 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     7141 cggccatttt ccaccatgat attcggcaag caggcatcgc cgtgggtcac gacgagatcc
     7201 tcgccgtcgg gcatccgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     7261 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     7321 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     7381 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     7441 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     7501 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     7561 tcttggagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     7621 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     7681 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     7741 atcatgcctc gatcgagttg agagtgaata tgagactcta attggatacc gaggggaatt
     7801 tatggaacgt cagtggagca tttttgacaa gaaatatttg ctagctgata gtgaccttag
     7861 gcgacttttg aacgcgcaat aatggtttct gacgtatgtg cttagctcat taaactccag
     7921 aaacccgcgg ctgagtggct ccttcaacgt tgcggttctg tcagttccaa acgtaaaacg
     7981 gcttgtcccg cgtcatcggc gggggtcata acgtgactcc cttaattctc cgctcatgat
     8041 cgataattcg cggtacccgg ggatcctagg gataacaggg taatatccct attgtcccat
     8101 taaaatagga aagtctggta actcaggaca ctgtatctgc tacgctgttt ataagcttgg
     8161 cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat
     8221 atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt
     8281 acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct
     8341 ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga
     8401 aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt
     8461 ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta taaaagagag
     8521 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg
     8581 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtcccccg tgaactttac
     8641 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     8701 ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     8761 aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat acacagccag
     8821 taagctttaa ggtttcagtc acactacgga tacttttaca acgggagcag ttattcaatt
     8881 accctgttat ccctacagct tagcttgagc ttggatcaga ttgtcgtttc ccgccttcag
     8941 tttaaactat cagtgtttga caggatatat tggcgggtaa a
//
