LOCUS       40924_34396        8042 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pPgrac-cel8A, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8042)
  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  JOURNAL   Unpublished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 8042)
  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 8042)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8042)
  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            "Unpublished"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..8042
                     /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)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1699..1803)
                     /label=AmpR promoter
     CDS             complement(1832..2116)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /protein_id="AXF54477.1"
                     /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="AXF54478.1"
                     /translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            complement(2515..3276)
                     /gene="kanR"
                     /label=kanR
     CDS             complement(3448..4449)
                     /codon_start=1
                     /label=repB
                     /note="RepB replication protein"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     CDS             complement(4905..5984)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    6368..6392
                     /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)."
     CDS             6419..6502
                     /codon_start=1
                     /gene="SPLipB-Cel8A-His6"
                     /product="LipB signal peptide"
                     /label=SPLipB-Cel8A-His6
                     /protein_id="AXF54480.1"
                     /translation="MKKVLMAFIICLSLILSVLAAPPSGAKA"
     CDS             7862..7879
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
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 ctagtgagct caggccttaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4921 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4981 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     5041 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     5101 aggcgaaaat cctgtttgat ggtggttgac ggcgggatat aacatgagct gtcttcggta
     5161 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     5221 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     5281 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     5341 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     5401 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     5461 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     5521 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     5581 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     5641 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     5701 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     5761 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     5821 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     5881 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     5941 acaccggcat actctgcgac atcgtataac gttactggtt tcatcaaaat cgtctccctc
     6001 cgtttgaata tttgattgat cgtaaccaga tgaagcactc tttccactat ccctacagtg
     6061 ttatggcttg aacaatcacg aaacaataat tggtacgtac gatctttcag ccgactcaaa
     6121 catcaaatct tacaaatgta gtctttgaaa gtattacata tgtaagattt aaatgcaacc
     6181 gttttttcgg aaggaaatga tgacctcgtt tccaccggaa ttagcttggt accagctatt
     6241 gtaacataat cggtacgggg gtgaaaaagc taacggaaaa gggagcggaa aagaatgatg
     6301 taagcgtgaa aaatttttta tcttatcact tgaaattgga agggagattc tttattataa
     6361 gaattgtgga attgtgagcg gataacaatt cccaattaaa ggaggaagga tcacgcgtat
     6421 gaaaaaagta cttatggcat tcattatttg tttatcgctg attctatctg ttttagccgc
     6481 tccgccgtct ggcgcaaaag ctgcggccgg tgcacatatg gccggagttc cttttaacac
     6541 gaagtatcct tatgggccaa catcaattgc tgacaatcaa tctgaggtta cagccatgtt
     6601 aaaagctgaa tgggaagatt ggaaaagtaa acgtatcacc tctaatggtg caggcggcta
     6661 caaaagagtg caaagagatg cgtcaacaaa ttacgatact gtatctgaag gaatgggata
     6721 tggtctgtta ttagcggttt gctttaacga acaagcccta tttgacgact tataccgata
     6781 tgtaaaatct catttcaacg gcaatggatt aatgcattgg catattgatg caaataacaa
     6841 cgtaacctca catgatggag gtgatggagc agctacagat gcagacgagg atattgcact
     6901 agctctgata tttgctgaca aattatgggg aagttctgga gctattaatt acgggcaaga
     6961 agcacgtaca cttattaaca atctgtataa tcactgtgta gaacatggct cttatgtctt
     7021 gaaaccagga gatcgttggg gtggtagcag cgtgacaaac ccatcatatt ttgcacctgc
     7081 atggtataaa gtgtatgctc agtatactgg agatacccgt tggaatcagg ttgcagacaa
     7141 atgctatcag atagtagaag aagtgaagaa gtacaacaat ggcactggtc ttgttcccga
     7201 ttggtgtaca gcatctggca ctccagcttc aggacagagt tacgattata agtatgacgc
     7261 tactcgatat gggtggagaa ctgccgttga ttattcatgg tttggagatc aacgtgcaaa
     7321 agcgaattgt gatatgttaa cgaaattctt tgctagagat ggtgcgaaag gtattgtcga
     7381 tggctacaca attcaaggct ctaaaatcag caataatcac aatgcttctt tcattggacc
     7441 tgtagctgct gcttcaatga cgggatacga tcttaacttt gccaaagaac tatatcgcga
     7501 aacagttgca gtaaaggata gtgagtatta tgggtactat ggaaacagtt tacgcctttt
     7561 aaccctttta tatattacgg gtaattttcc caatccttta agcgatttat ctgggcaacc
     7621 tacgcctcca tctaatccga caccgtcttt acctcctcaa gttgtgtatg gtgatgtaaa
     7681 tggtgacgga aatgtcaact ctacagacct aacaatgttg aaacgttatc ttcttaagtc
     7741 agttacgaac atcaaccgtg aagcagccga tgtaaatcga gatggagcga ttaatagcag
     7801 tgatatgacg atattgaaac gctacttgat caaatctatt ccgcatttac catattctag
     7861 acatcaccat catcaccact aatgcggtag tttatcacag ttaaattgct aacgcagtca
     7921 ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg tcaccctgga
     7981 tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctatttcact ttttgcattc
     8041 ta
//
