LOCUS      pB7WG_ASX               8723 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Plant expression vector for multigene cloning.
ACCESSION   .
VERSION     .
KEYWORDS    pB7WG_ASX
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8723)
  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. 2018 Jan 4;13(1):e0190526. doi: 
            10.1371/journal.pone.0190526. eCollection 2018.
  PUBMED    29300787
REFERENCE   2  (bases 1 to 8723)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..8723
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1238..1867
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2296..3369
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3435..3629
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3865..3887)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3973..4113
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4028..4047
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4128..4145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4299..4887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4379..4398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5133..5924)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6445..6469
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      6562..6814
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(6834..7385)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(7427..7606)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             8136..8441
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     8320..8339
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    8606..8630
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 cagggttccc ctcgggatca aagtacttta aagtacttta aagtacttta aagtactttg
       61 atccaacccc tccgctgcta tagtgcagtc ggcttctgac gttcagtgca gccgtcttct
      121 gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct gcccttttcc
      181 tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga ctagaaccgg
      241 agacattacg ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc
      301 accgacgacc aggacttgac caaccaacgg gccgaactgc acgcggccgg ctgcaccaag
      361 ctgttttccg agaagatcac cggcaccagg cgcgaccgcc cggagctggc caggatgctt
      421 gaccacctac gccctggcga cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc
      481 acccgcgacc tactggacat tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc
      541 ctggcagagc cgtgggccga caccaccacg ccggccggcc gcatggtgtt gaccgtgttc
      601 gccggcattg ccgagttcga gcgttcccta atcatcgacc gcacccggag cgggcgcgag
      661 gccgccaagg cccgaggcgt gaagtttggc ccccgcccta ccctcacccc ggcacagatc
      721 gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg
      781 cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc gcagcgagga agtgacgccc
      841 accgaggcca ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc cgacgccctg
      901 gcggccgccg agaatgaacg ccaagaggaa caagcatgaa accgcaccag gacggccagg
      961 acgaaccgtt tttcattacc gaagagatcg aggcggagat gatcgcggcc gggtacgtgt
     1021 tcgagccgcc cgcgcacgtc tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg
     1081 atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc
     1141 taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga
     1201 tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa
     1261 ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact
     1321 cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc
     1381 ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg
     1441 cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc
     1501 ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag
     1561 cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt
     1621 cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat
     1681 cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat
     1741 cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga
     1801 acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact
     1861 catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc
     1921 cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc
     1981 atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg
     2041 gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag
     2101 taaatgagca aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga
     2161 aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg
     2221 ttggccaggc gtaagcggct gggttgtctg ccggccctgc aatggcactg gaacccccaa
     2281 gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg
     2341 ctgggtgatg acctggtgga gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc
     2401 gaggcagaag cacgccccgg tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa
     2461 tcccggcaac cgccggcagc cggtgcgccg tcgattagga agccgcccaa gggcgacgag
     2521 caaccagatt ttttcgttcc gatgctctat gacgtgggca cccgcgatag tcgcagcatc
     2581 atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc
     2641 tacgagcttc cagacgggca cgtagaggtt tccgcagggc cggccggcat ggccagtgtg
     2701 tgggattacg acctggtact gatggcggtt tcccatctaa ccgaatccat gaaccgatac
     2761 cgggaaggga agggagacaa gcccggccgc gtgttccgtc cacacgttgc ggacgtactc
     2821 aagttctgcc ggcgagccga tggcggaaag cagaaagacg acctggtaga aacctgcatt
     2881 cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga aggccaagaa cggccgcctg
     2941 gtgacggtat ccgagggtga agccttgatt agccgctaca agatcgtaaa gagcgaaacc
     3001 gggcggccgg agtacatcga gatcgagcta gctgattgga tgtaccgcga gatcacagaa
     3061 ggcaagaacc cggacgtgct gacggttcac cccgattact ttttgatcga tcccggcatc
     3121 ggccgttttc tctaccgcct ggcacgccgc gccgcaggca aggcagaagc cagatggttg
     3181 ttcaagacga tctacgaacg cagtggcagc gccggagagt tcaagaagtt ctgtttcacc
     3241 gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg atttgaagga ggaggcgggg
     3301 caggctggcc cgatcctagt catgcgctac cgcaacctga tcgagggcga agcatccgcc
     3361 ggttcctaat gtacggagca gatgctaggg caaattgccc tagcagggga aaaaggtcga
     3421 aaaggtctct ttcctgtgga tagcacgtac attgggaacc caaagccgta cattgggaac
     3481 cggaacccgt acattgggaa cccaaagccg tacattggga accggtcaca catgtaagtg
     3541 actgatataa aagagaaaaa aggcgatttt tccgcctaaa actctttaaa acttattaaa
     3601 actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg
     3661 caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct
     3721 atcgcggccg ctggccgctc aaaaatggct ggcctacggc caggcaatct accagggcgc
     3781 ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac atcaaggcac cctgcctcgc
     3841 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3901 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3961 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     4021 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     4081 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     4141 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4201 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4261 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4321 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4381 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4441 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4501 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4561 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4621 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4681 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     4741 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4801 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4861 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4921 gctcagtgga acgaaaactc acgttaaggg attttggtca tgcatgatat atctcccaat
     4981 ttgtgtaggg cttattatgc acgcttaaaa ataataaaag cagacttgac ctgatagttt
     5041 ggctgtgagc aattatgtgc ttagtgcatc taacgcttga gttaagccgc gccgcgaagc
     5101 ggcgtcggct tgaacgaatt tctagctaga cattatttgc cgactacctt ggtgatctcg
     5161 cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct
     5221 tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg ggccggcagg
     5281 cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg
     5341 taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc
     5401 gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga
     5461 tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc
     5521 agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca
     5581 ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg
     5641 tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa
     5701 gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg
     5761 gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag
     5821 ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc
     5881 tctgatagtt gagtcgatac ttcggcgatc accgcttccc ccatgatgtt taactttgtt
     5941 ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     6001 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacccc aaaaaaacag
     6061 tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt
     6121 ctggaccagt tgcgtgagcg catacgctac ttgcattaca gcttacgaac cgaacaggct
     6181 tatgtccact gggttcgtgc ccgaattgat cacaggcagc aacgctctgt catcgttaca
     6241 atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt
     6301 tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct
     6361 gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
     6421 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct
     6481 tagacaactt aataacacat tgcggacgtt tttaatgtac tgaattaacg ccgaattgaa
     6541 ttatcagctt gcatgccggt cgatctagta acatagatga caccgcgcgc gataatttat
     6601 cctagtttgc gcgctatatt ttgttttcta tcgcgtatta aatgtataat tgcgggactc
     6661 taatcataaa aacccatctc ataaataacg tcatgcatta catgttaatt attacatgct
     6721 taacgtaatt caacagaaat tatatgataa tcatcgcaag accggcaaca ggattcaatc
     6781 ttaagaaact ttattgccaa atgtttgaac gatctgcttg actctagggg tcatcagatt
     6841 tcggtgacgg gcaggaccgg acggggcggc accggcaggc tgaagtccag ctgccagaaa
     6901 cccacgtcat gccagttccc gtgcttgaag ccggccgccc gcagcatgcc gcggggggca
     6961 tatccgagcg cctcgtgcat gcgcacgctc gggtcgttgg gcagcccgat gacagcgacc
     7021 acgctcttga agccctgtgc ctccagggac ttcagcaggt gggtgtagag cgtggagccc
     7081 agtcccgtcc gctggtggcg gggggagacg tacacggtcg actcggccgt ccagtcgtag
     7141 gcgttgcgtg ccttccaggg acccgcgtag gcgatgccgg cgacctcgcc gtccacctcg
     7201 gcgacgagcc agggatagcg ctcccgcaga cggacgaggt cgtccgtcca ctcctgcggt
     7261 tcctgcggct cggtacggaa gttgaccgtg cttgtctcga tgtagtggtt gacgatggtg
     7321 cagaccgccg gcatgtccgc ctcggtggca cggcggatgt cggccgggcg tcgttctggg
     7381 ctcatggtag atcccctcga tcgagttgag agtgaatatg agactctaat tggataccga
     7441 ggggaattta tggaacgtca gtggagcatt tttgacaaga aatatttgct agctgatagt
     7501 gaccttaggc gacttttgaa cgcgcaataa tggtttctga cgtatgtgct tagctcatta
     7561 aactccagaa acccgcggct cagtggctcc ttcaacgttg cggttctgtc agttccaaac
     7621 gtaaaacggc ttgtcccgcg tcatcggcgg gggtcataac gtgactccct taattctccg
     7681 ctcatgatcg ataattcgcg gtacccgggg atcctaggga taacagggta atatccctat
     7741 tgtcccatta aaataggaaa gtctggtaac tcaggacact gtatctgcta cgctgtttat
     7801 aagcttggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     7861 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
     7921 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     7981 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     8041 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     8101 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     8161 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     8221 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtcccccgtg
     8281 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     8341 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     8401 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     8461 acagccagta agctttaagg tttcagtcac actacggata cttttacaac gggagcagtt
     8521 attcaattac cctgttatcc ctacagctta gcttgagctt ggatcagatt gtcgtttccc
     8581 gccttcagtt taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa
     8641 agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc
     8701 catttgtatg tgcatgccaa cca
//