LOCUS       40924_35718        8084 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pPxylA+-cel8A, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8084)
  AUTHORS   Heinze S, Zimmermann K, Ludwig C, Heinzlmeir S, Schwarz WH, Zverlov 
            VV, Liebl W, Kornberger P.
  TITLE     Efficient secretory heterologous enzyme production in the novel host
            Paenibacillus polymyxa by usage of its promoter sequences
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8084)
  AUTHORS   Heinze S, Zimmermann K, Ludwig C, Heinzlmeir S, Schwarz WH, Zverlov 
            VV, Liebl W, Kornberger P.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAY-2018) Department for Microbiology, Technical 
            University of Munich, Emil-Ramann-Str. 4, Freising, Bavaria 85354, 
            Germany
REFERENCE   3  (bases 1 to 8084)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8084)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-MAY-2018) Department for Microbiology, Technical University of 
            Munich, Emil-Ramann-Str. 4, Freising, Bavaria 85354, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(79..667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(841..1698)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1699..1803)
                     /label=AmpR promoter
     CDS             complement(1832..2116)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /protein_id="AXF54489.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGQQKAHYVNHHPNQVFWGRGAVKH"
     gene            complement(1832..2116)
                     /gene="traJ"
                     /label=traJ
     oriT            complement(2149..2258)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2515..3276)
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /label=kanR
                     /note="Kanamycin Resistance"
                     /protein_id="AXF54490.1"
                     /translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            complement(2515..3276)
                     /gene="kanR"
                     /label=kanR
     CDS             complement(3448..4449)
                     /label=repB
                     /note="RepB replication protein"
     CDS             complement(5063..6229)
                     /codon_start=1
                     /gene="xylR"
                     /product="XylR"
                     /label=xylR
                     /note="XylR from B. megaterium xyl-operon"
                     /protein_id="AXF54491.1"
                     /translation="MVIIQIADQALVKKMNQKLILDEILKNSPVSRATLSEITGLNKST
                     VSSQVNTLLEKDFIFEIGAGQSRGGRRPVMLVFNKNAGYSIGIDIGVDYLNGILTDLEG
                     NIILEKTSDLSSSSASEVKEILFALIHGFVTHMPESPYGLVGIGICVPGLVDRHQQIIF
                     MPNLNWNIKDLQFLIESEFNVPVFVENEANAGAYGEKVFGMTKNYENIVYISINIGIGT
                     GLVINNELYKGVQGFSGEMGHMTIDFNGPKCSCGNRGCWELYASEKALLASLSKEEKNI
                     SRKEIVERANKNDVEMLNALQNFGFYIGIGLTNILNTFDIEAVILRNHIIESHPIVLNT
                     IKNEVSSRVHSHLDNKCELLPSSLGKNAPALGAVSIVIDSFLSVTPIS"
     gene            complement(5063..6229)
                     /gene="xylR"
                     /label=xylR
     regulatory      6333..6367
                     /note="PxylA, xylose inducible promoter, derived from
                     pHIS1525"
                     /regulatory_class="promoter"
     misc_feature    6369..6397
                     /label=XylR binding site
                     /note="XylR binding site"
     regulatory      6438..6447
                     /note="rbs+, ribosome binding site optimized for B.
                     megaterium"
                     /regulatory_class="ribosome_binding_site"
     CDS             6461..6544
                     /codon_start=1
                     /gene="SPLipB-Cel8A-His6"
                     /product="LipB signal peptide"
                     /label=SPLipB-Cel8A-His6
                     /protein_id="AXF54492.1"
                     /translation="MKKVLMAFIICLSLILSVLAAPPSGAKA"
     CDS             7904..7921
                     /label=6xHis
                     /note="6xHis affinity tag"
ORIGIN
        1 caaactgcat aactattatg tacgagcaaa aggccagcaa aaggccagga accgtaaaaa
       61 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      481 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1141 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1441 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1801 gcgcacattt ccccgaaaag tgccacctga cttagtgctt tacggcacct cgaccccaaa
     1861 aaacttgatt agggtgatgg ttcacgtagt gggccttttg ctggcccagg tcgccattga
     1921 tgcgggccag ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc
     1981 cgacggccag caggtaggcc gacaggctca tgcctgccgc cgccgccttt tcctcaatcg
     2041 ctcttcgttc gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct
     2101 tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc
     2161 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
     2221 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
     2281 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
     2341 ataccgaaaa aatcgctata atgaccccga agcgtctaag aaaccattat tatcatgaca
     2401 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     2461 ggtgaaaacc tctgacagta accaacatga ttaacaatta ttagaggtca tcgttcaaaa
     2521 tggtatgcgt tttgacacat ccactatata tccgtgtcgt tctgtccact cctgaatccc
     2581 attccagaaa ttctctagcg attccagaag tttctcagag tcggaaagtt gaccagacat
     2641 tacgaactgg cacagatggt cataacctga aggaagatct gattgcttaa ctgcttcagt
     2701 taagaccgaa gcgctcgtcg tataacagat gcgatgatgc agaccaatca acatggcacc
     2761 tgccattgct acctgtacag tcaaggatgg tagaaatgtt gtcggtcctt gcacacgaat
     2821 attacgccat ttgcctgcat attcaaacag ctcttctacg ataagggcac aaatcgcatc
     2881 gtggaacgtt tgggcttcta ccgatttagc agtttgatac actttctcta agtatccacc
     2941 tgaatcataa atcggcaaaa tagagaaaaa ttgaccatgt gtaagcggcc aatctgattc
     3001 cacctgagat gcataatcta gtagaatctc ttcgctatca aaattcactt ccaccttcca
     3061 ctcaccggtt gtccattcat ggctgaactc tgcttcctct gttgacatga cacacatcat
     3121 ctcaatatcc gaatagggcc catcagtctg acgaccaaga gagccataaa caccaatagc
     3181 cttaacatca tccccatatt tatccaatat tcgttcctta atttcatgaa caatcttcat
     3241 tctttcttct ctagtcatta ttattggtcc attcactatt ctcattccct tttcagataa
     3301 ttttagattt gcttttctaa ataagaatat ttggagagca ccgttcttat tcagctatta
     3361 ataactcgtc ttcctaagca tccttcaatc cttttaataa caattatagc atctaatctt
     3421 caacaaactg gcccgtttgt tgaactactc tttaataaaa taatttttcc gttcccaatt
     3481 ccacattgca ataatagaaa atccatcttc atcggctttt tcgtcatcat ctgtatgaat
     3541 caaatcgcct tcttctgtgt catcaaggtt taatttttta tgtatttctt ttaacaaacc
     3601 accataggag attaaccttt tacggtgtaa accttcctcc aaatcagaca aacgtttcaa
     3661 attcttttct tcatcatcgg tcataaaatc cgtatccttt acaggatatt ttgcagtttc
     3721 gtcaattgcc gattgtatat ccgatttata tttatttttc ggtcgaatca tttgaacttt
     3781 tacatttgga tcatagtcta atttcattgc ctttttccaa aattgaatcc attgtttttg
     3841 attcacgtag ttttctgtat tcttaaaata agttggttcc acacatacca atacatgcat
     3901 gtgctgatta taagaattat ctttattatt tattgtcact tccgttgcac gcataaaacc
     3961 aacaagattt ttattaattt ttttatattg catcattcgg cgaaatcctt gagccatatc
     4021 tgacaaactc ttatttaatt cttcgccatc ataaacattt ttaactgtta atgtgagaaa
     4081 caaccaacga actgttggct tttgtttaat aacttcagca acaacctttt gtgactgaat
     4141 gccatgtttc attgctctcc tccagttgca cattggacaa agcctggatt tacaaaacca
     4201 cactcgatac aactttcttt cgcctgtttc acgattttgt ttatactcta atatttcagc
     4261 acaatctttt actctttcag cctttttaaa ttcaagaata tgcagaagtt caaagtaatc
     4321 aacattagcg attttctttt ctctccatgg tctcactttt ccactttttg tcttgtccac
     4381 taaaaccctt gatttttcat ctgaataaat gctactatta ggacacataa tattaaaaga
     4441 aacccccatc tatttagtta tttgtttagt cacttataac tttaacagat ggggtttttc
     4501 tgtgcaacca attttaaggg ttttcaatac tttaaaacac atacatacca acacttcaac
     4561 gcacctttca gcaactaaaa taaaaatgac gttatttcta tatgtatcaa gataagaaag
     4621 aacaagttca aaaccatcaa aaaaagacac cttttcaggt gcttttttta ttttataaac
     4681 tcattccctg atctcgactt cgttcttttt ttacctctcg gttatgagtt agttcaaatt
     4741 cgttcttttt aggttctaaa tcgtgttttt cttggaattg tgctgtttta tcctttacct
     4801 tgtctacaaa ccccttaaaa acgtttttaa aggcttttaa gccgtctgta cgttcctaaa
     4861 ctagtaacat ataaacagcc agttgccgtt atgataggtg actggctgca tgggatgaaa
     4921 aggtgagggt ggagacagac ataacactct taatagaaga gggtaattct ttctctttta
     4981 tagaaaatca attaattgaa agtagctcct tcattcttaa gatcaacgtg atataggttt
     5041 gctaaccttt gcgttcactt aactaactta taggggtaac acttaaaaaa gaatcaataa
     5101 cgatagaaac cgctcctaaa gcaggtgcat tttttcctaa cgaagaaggc aatagttcac
     5161 atttattgtc taaatgagaa tggactctag aagaaacttc gtttttaatc gtatttaaaa
     5221 caatgggatg agattcaatt atatgatttc tcaagataac agcttctata tcaaatgtat
     5281 taaggatatt ggttaatcca attccgatat aaaagccaaa gttttgaagt gcatttaaca
     5341 tttctacatc atttttattt gcgcgttcca caatctcttt tcgagaaata ttcttttctt
     5401 ctttagagag cgaagccagt aacgcttttt cagaagcata taattcccaa cagcctcgat
     5461 ttccacagct gcatttgggt ccattaaaat ctatcgtcat atgacccatt tccccagaaa
     5521 aaccctgaac acctttatac aattcgttgt taataacaag tccagttcca attccgatat
     5581 taatactgat gtaaacgatg ttttcatagt tttttgtcat accaaatact ttttcaccgt
     5641 atgctcctgc attagcttca ttttcaacaa aaaccggaac attaaactca ctctcaatta
     5701 aaaactgcaa atctttgata ttccaattta agttaggcat gaaaataatt tgctgatgac
     5761 gatctacaag gcctggaaca caaattccta ttccgactag accataaggg gactcaggca
     5821 tatgggttac aaaaccatga ataagtgcaa ataaaatctc ttttacttca ctagcggaag
     5881 aactagacaa gtcagaagtc ttctcgagaa taatatttcc ttctaagtcg gttagaattc
     5941 cgttaagata gtcgactcct atatcaatac caatcgagta gcctgcattc ttattaaaaa
     6001 caagcattac aggtcttctg ccgcctctag attgccctgc cccaatttca aaaataaaat
     6061 ctttttcaag cagtgtattt acttgagagg agacagtaga cttgtttaat cctgtaatct
     6121 cagagagagt tgccctggag acaggggagt tcttcaaaat ttcatctaat attaattttt
     6181 gattcatttt ttttactaaa gcttgatctg caatttgaat aataaccact cctttgttta
     6241 tccaccgaac taagttggtg ttttttgaag cttgaattag atatttaaaa gtatcatatc
     6301 taatattata actaaatttt ctaaaaaaaa cattgaaata aacatttatt ttgtatatga
     6361 tgagataaag ttagtttatt ggataaacaa actaactcaa ttaagatagt tgatggataa
     6421 acttgttcac ttaaatcaag gaggtgaatg tacaacgcgt atgaaaaaag tacttatggc
     6481 attcattatt tgtttatcgc tgattctatc tgttttagcc gctccgccgt ctggcgcaaa
     6541 agctgcggcc ggtgcacata tggccggagt tccttttaac acgaagtatc cttatgggcc
     6601 aacatcaatt gctgacaatc aatctgaggt tacagccatg ttaaaagctg aatgggaaga
     6661 ttggaaaagt aaacgtatca cctctaatgg tgcaggcggc tacaaaagag tgcaaagaga
     6721 tgcgtcaaca aattacgata ctgtatctga aggaatggga tatggtctgt tattagcggt
     6781 ttgctttaac gaacaagccc tatttgacga cttataccga tatgtaaaat ctcatttcaa
     6841 cggcaatgga ttaatgcatt ggcatattga tgcaaataac aacgtaacct cacatgatgg
     6901 aggtgatgga gcagctacag atgcagacga ggatattgca ctagctctga tatttgctga
     6961 caaattatgg ggaagttctg gagctattaa ttacgggcaa gaagcacgta cacttattaa
     7021 caatctgtat aatcactgtg tagaacatgg ctcttatgtc ttgaaaccag gagatcgttg
     7081 gggtggtagc agcgtgacaa acccatcata ttttgcacct gcatggtata aagtgtatgc
     7141 tcagtatact ggagataccc gttggaatca ggttgcagac aaatgctatc agatagtaga
     7201 agaagtgaag aagtacaaca atggcactgg tcttgttccc gattggtgta cagcatctgg
     7261 cactccagct tcaggacaga gttacgatta taagtatgac gctactcgat atgggtggag
     7321 aactgccgtt gattattcat ggtttggaga tcaacgtgca aaagcgaatt gtgatatgtt
     7381 aacgaaattc tttgctagag atggtgcgaa aggtattgtc gatggctaca caattcaagg
     7441 ctctaaaatc agcaataatc acaatgcttc tttcattgga cctgtagctg ctgcttcaat
     7501 gacgggatac gatcttaact ttgccaaaga actatatcgc gaaacagttg cagtaaagga
     7561 tagtgagtat tatgggtact atggaaacag tttacgcctt ttaacccttt tatatattac
     7621 gggtaatttt cccaatcctt taagcgattt atctgggcaa cctacgcctc catctaatcc
     7681 gacaccgtct ttacctcctc aagttgtgta tggtgatgta aatggtgacg gaaatgtcaa
     7741 ctctacagac ctaacaatgt tgaaacgtta tcttcttaag tcagttacga acatcaaccg
     7801 tgaagcagcc gatgtaaatc gagatggagc gattaatagc agtgatatga cgatattgaa
     7861 acgctacttg atcaaatcta ttccgcattt accatattct agacatcacc atcatcacca
     7921 ctaatgcggt agtttatcac agttaaattg ctaacgcagt caggcaccgt gtatgaaatc
     7981 taacaatgcg ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag gcataggctt
     8041 ggttatgccg gtactgccgg gcctatttca ctttttgcat tcta
//
