LOCUS       40924_24114        6838 bp DNA     circular SYN 18-DEC-2018
DEFINITION  GFP expression vector pHapII, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6838)
  AUTHORS   Zhang N, Wu K, He X, Li S-Q., Zhang Z-H., Shen B, Yang Y-M., Zhang 
            R-F., Huang Q-W., Shen Q-R.
  TITLE     A new bioorganic fertilizer can effectively control banana wilt by 
            strong colonization with Bacillus subtilis N11
  JOURNAL   Plant Soil 344 (1-2), 87-97 (2011)
REFERENCE   2  (bases 1 to 6838)
  AUTHORS   Zhang Z, Tian W, Liu D.
  TITLE     Development of a GFP vector system in Bacillus under the control of 
            the constitutive promoter HpaII
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6838)
  AUTHORS   Zhang Z, Tian W, Liu D, Zhou T, Wu Y, Liu Y, Xu L.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-APR-2010) Jiang Su Provincial Key Lab for Organic 
            Solid Waste Utilization, Nanjing Agricultural University, Wei Gang 
            #1, Nan jing, Jiang Su 210095, China
REFERENCE   4  (bases 1 to 6838)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6838)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Soil 
            344 (1-2), 87-97 (2011)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (26-APR-2010) Jiang Su Provincial Key Lab for Organic Solid Waste 
            Utilization, Nanjing Agricultural University, Wei Gang #1, Nan jing,
            Jiang Su 210095, China"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             814..1815
                     /codon_start=1
                     /label=repB
                     /note="RepB replication protein"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     CDS             1987..2748
                     /codon_start=1
                     /product="kanamycin nucleotidyltranferase"
                     /label=kanamycin nucleotidyltranferase
                     /note="KM"
                     /protein_id="ADK73494.1"
                     /translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     CDS             2965..3216
                     /codon_start=1
                     /product="ble"
                     /label=ble
                     /protein_id="ADK73495.1"
                     /translation="MRMLQSIPALPVGDIKKSIGFYWDKVGFTLVHHEDGFAVLMCNEV
                     RIHLWEASDEGWRLVVMIHRFVQVRSRLLLVLLVAALK"
     CDS             3873..4586
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4617..4633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4641..4657)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4665..4695)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4710..4731)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5019..5607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5781..6638)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6639..6743)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ccttaaggaa cgtacagacg
      421 gcttaaaagc ctttaaaaac gtttttaagg ggtttgtaga caaggtaaag gataaaacag
      481 cacaattcca agaaaaacac gatttagaac ctaaaaagaa cgaatttgaa ctaactcata
      541 accgagaggt aaaaaaagaa cgaagtcgag atcagggaat gagtttataa aataaaaaaa
      601 gcacctgaaa aggtgtcttt ttttgatggt tttgaacttg ttctttctta tcttgataca
      661 tatagaaata acgtcatttt tattttagtt gctgaaaggt gcgttgaagt gttggtatgt
      721 atgtgtttta aagtattgaa aacccttaaa attggttgca cagaaaaacc ccatctgtta
      781 aagttataag tgactaaaca aataactaaa tagatggggg tttcttttaa tattatgtgt
      841 cctaatagta gcatttattc agatgaaaaa tcaagggttt tagtggacaa gacaaaaagt
      901 ggaaaagtga gaccatggag agaaaagaaa atcgctaatg ttgattactt tgaacttctg
      961 catattcttg aatttaaaaa ggctgaaaga gtaaaagatt gtgctgaaat attagagtat
     1021 aaacaaaatc gtgaaacagg cgaaagaaag ttgtatcgag tgtggttttg taaatccagg
     1081 ctttgtccaa tgtgcaactg gaggagagca atgaaacatg gcattcagtc acaaaaggtt
     1141 gttgctgaag ttattaaaca aaagccaaca gttcgttggt tgtttctcac attaacagtt
     1201 aaaaatgttt atgatggcga agaattaaat aagagtttgt cagatatggc tcaaggattt
     1261 cgccgaatga tgcaatataa aaaaattaat aaaaatcttg ttggttttat gcgtgcaacg
     1321 gaagtgacaa taaataataa agataattct tataatcagc acatgcatgt attggtatgt
     1381 gtggaaccaa cttattttaa gaatacagaa aactacgtga atcaaaaaca atggattcaa
     1441 ttttggaaaa aggcaatgaa attagactat gatccaaatg taaaagttca aatgattcga
     1501 ccgaaaaata aatataaatc ggatatacaa tcggcaattg acgaaactgc aaaatatcct
     1561 gtaaaggata cggattttat gaccgatgat gaagaaaaga atttgaaacg tttgtctgat
     1621 ttggaggaag gtttacaccg taaaaggtta atctcctatg gtggtttgtt aaaagaaata
     1681 cataaaaaat taaaccttga tgacacagaa gaaggcgatt tgattcatac agatgatgac
     1741 gaaaaagccg atgaagatgg attttctatt attgcaatgt ggaattggga acggaaaaat
     1801 tattttatta aagagtagtt caacaaacgg gccagtttgt tgaagattag atgctataat
     1861 tgttattaaa aggattgaag gatgcttagg aagacgagtt attaatagct gaataagaac
     1921 ggtgctctcc aaatattctt atttagaaaa gcaaatctaa aattatctga aaagggaatg
     1981 agaataatga atggaccaat aataatgact agagaagaaa gaatgaagat tgttcatgaa
     2041 attaaggaac gaatattgga taaatatggg gatgatgtta aggctattgg tgtttatggc
     2101 tctcttggtc gtcagactga tgggccctat tcggatattg agatgatgtg tgtcatgtca
     2161 acagaggaag cagagttcag ccatgaatgg acaaccggtg agtggaaggt ggaagtgaat
     2221 tttgatagcg aagagattct actagattat gcatctcagg tggaatcaga ttggccgctt
     2281 acacatggtc aatttttctc tattttgccg atttatgatt caggtggata cttagagaaa
     2341 gtgtatcaaa ctgctaaatc ggtagaagcc caaacgttcc acgatgcgat ttgtgccctt
     2401 atcgtagaag agctgtttga atatgcaggc aaatggcgta atattcgtgt gcaaggaccg
     2461 acaacatttc taccatcctt gactgtacag gtagcaatgg caggtgccat gttgattggt
     2521 ctgcatcatc gcatctgtta tacgacgagc gcttcggtct taactgaagc agttaagcaa
     2581 tcagatcttc cttcaggtta tgaccatctg tgccagttcg taatgtctgg tcaactttcc
     2641 gactctgaga aacttctgga atcgctagag aatttctgga atgggattca ggagtggaca
     2701 gaacgacacg gatatatagt ggatgtgtca aaacgcatac cattttgaac gatgacctct
     2761 aataattgtt aatcatgttg gttacgtatt tattaacttc tcctagtatt agtaattatc
     2821 atggctgtca tggcgcatta acggaataaa gggtgtgctt aaatcgggcc attttgcgta
     2881 ataagaaaaa ggattaatta tgagcgaatt gaattaataa taaggtaata gatttacatt
     2941 agaaaatgaa aggggatttt atgcatgaga atgttacagt ctatcccggc attgccagtc
     3001 ggggatatta aaaagagtat aggtttttat tgggataaag taggtttcac tttggttcac
     3061 catgaagatg gattcgcagt tctaatgtgt aatgaggttc ggattcatct atgggaggca
     3121 agtgatgaag gctggcgcct cgtagtaatg attcaccggt ttgtacaggt gcggagtcgt
     3181 ttattgctgg tactgctagt tgccgcattg aagtagaggg aattgatgaa ttatatcaac
     3241 atattaagcc tttgggcatt ttgcacccca atacatcatt aaaagatcag tggtgggatg
     3301 aacgagactt tgcagtaatt gatcccgaca acaatttgat tagctttttt caacaaataa
     3361 aaagctaaaa tctattatta atctgttcag caatcgggcg cgattgctga ataaaagata
     3421 cgagagacct ctcttgtatc ttttttattt tgagtggttt tgtccgttac actagaaaac
     3481 cgaaagacaa taaaaatttt attcttgctg agtctggctt tcggtaagct agacaaaacg
     3541 gacaaaataa aaattggcaa gggtttaaag gtggagattt tttgagtgat cttctcaaaa
     3601 aatactacct gtcccttgct gatttttaaa cgagcacgag agcaaaaccc ccctttgctg
     3661 aggtggcaga gggcaggttt ttttgtttct tttttctcgt aaaaaaaaga aaggtcttaa
     3721 aggttttatg gttttggtcg gcactgccgc gcctcgcaga gcacacactt tatgaatata
     3781 aagtatagtg tgttatactt tacttggaag tggttgccgg aaagagcgaa aatgcctcac
     3841 atttgtgcca cctaaaaagg agcgatttac atatgagtaa aggagaagaa cttttcactg
     3901 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     3961 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     4021 ctggaaaact acctgttcca tggccaacac ttgtcactac tttcggttat ggtgttcaat
     4081 gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt gccatgcccg
     4141 aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg
     4201 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
     4261 ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac tcacacaatg
     4321 tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca
     4381 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     4441 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
     4501 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     4561 cacatggcat ggatgaacta tacaaataaa tgctgcagaa gcttggcgta atcatggtca
     4621 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4681 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4741 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4801 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4861 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4921 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4981 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5041 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5101 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5161 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5221 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5281 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5341 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5401 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5461 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5521 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5581 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5641 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5701 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5761 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5821 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5881 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5941 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6001 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6061 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6121 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6181 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6241 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6301 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6361 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6421 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6481 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6541 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6601 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6661 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6721 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6781 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
