LOCUS       sequence-373044-        8784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector E. coli/B.meg.; encoding the RNA polymerase of the 
            phage SP6 under control of the xylose inducible promoter PxylA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8784)
  AUTHORS   Stammen S, Schuller F, Dietrich S, Gamer M, Biedendieck R, Jahn D
  TITLE     Application of Escherichia coli phage K1E DNA-dependent RNA 
            polymerase for in vitro RNA synthesis and in vivo protein production
            in Bacillus megaterium.
  JOURNAL   Appl Microbiol Biotechnol. 2010 Sep;88(2):529-39. doi: 
            10.1007/s00253-010-2732-y. Epub 2010 Jul 2.
  PUBMED    20596705
REFERENCE   2  (bases 1 to 8784)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1007/s00253-010-2732-y"; journalName: "Appl Microbiol 
            Biotechnol"; date: "2010-09"; volume: "88"; issue: "2"; pages: 
            "529-39"
COMMENT     Sequence Label: pSP6-RNAP
FEATURES             Location/Qualifiers
     source          1..8784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2586..2605)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2705..2727
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2765..2783)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2851..2955
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2956..3024,3025..3816)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3174..3193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3987..4575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4476..4495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4729..4746
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4863..4884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4899..4904,4905..4922,4923..4929)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4937..4953
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     4942..4964
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4961..4977
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4961..4977
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccggagatct tcgaactagt ttggaccatt tgtcatttcc ccctttgatt taagtgaaca
       61 agtttatcca tcaactatct taattgagtt agtttgttta tccaataaac taactttatc
      121 tcatcatata caaaataaat gtttatttca atgttttttt tagaaaattt agttataata
      181 ttagatatga tacttttaaa tatctaattc aagcttcaaa aaacaccaac ttagttcggt
      241 ggataaacaa aggagtggtt attattcaaa ttgcagatca agctttagta aaaaaaatga
      301 atcaaaaatt aatattagat gaaattttga agaactcccc tgtctccagg gcaactctct
      361 ctgagattac aggattaaac aagtctactg tctcctctca agtaaataca ctgcttgaaa
      421 aagattttat ttttgaaatt ggggcagggc aatctagagg cggcagaaga cctgtaatgc
      481 ttgtttttaa taagaatgca ggctactcga ttggtattga tataggagtc gactatctta
      541 acggaattct aaccgactta gaaggaaata ttattctcga gaagacttct gacttgtcta
      601 gttcttccgc tagtgaagta aaagagattt tatttgcact tattcatggt tttgtaaccc
      661 atatgcctga gtccccttat ggtctagtcg gaataggaat ttgtgttcca ggccttgtag
      721 atcgtcatca gcaaattatt ttcatgccta acttaaattg gaatatcaaa gatttgcagt
      781 ttttaattga gagtgagttt aatgttccgg tttttgttga aaatgaagct aatgcaggag
      841 catacggtga aaaagtattt ggtatgacaa aaaactatga aaacatcgtt tacatcagta
      901 ttaatatcgg aattggaact ggacttgtta ttaacaacga attgtataaa ggtgttcagg
      961 gtttttctgg ggaaatgggt catatgacga tagattttaa tggacccaaa tgcagctgtg
     1021 gaaatcgagg ctgttgggaa ttatatgctt ctgaaaaagc gttactggct tcgctctcta
     1081 aagaagaaaa gaatatttct cgaaaagaga ttgtggaacg cgcaaataaa aatgatgtag
     1141 aaatgttaaa tgcacttcaa aactttggct tttatatcgg aattggatta accaatatcc
     1201 ttaatacatt tgatatagaa gctgttatct tgagaaatca tataattgaa tctcatccca
     1261 ttgttttaaa tacgattaaa aacgaagttt cttctagagt ccattctcat ttagacaata
     1321 aatgtgaact attgccttct tcgttaggaa aaaatgcacc tgctttagga gcggtttcta
     1381 tcgttattga ttctttttta agtgttaccc ctataagtta gttaagtgaa cgcaaagggc
     1441 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagatcatt
     1501 atttagcgat gtcgcctact cctgaggacg atccggcaat agttaccctt attatcaaga
     1561 taagaaagaa aaggattttt cgctacgctc aaatccttta aaaaaacaca aaagaccaca
     1621 ttttttaatg tggtctttta ttcttcaact aaagcaccca ttagttcaac aaacgaaaat
     1681 tggataaagt gggatatttt taaaatatat atttatgtta cagtaatatt gacttttaaa
     1741 aaaggattga ttctaatgaa gaaagcagac aagtaagcct cctaaattca ctttagataa
     1801 aaatttagga ggcatatcaa atgaacttta ataaaattga tttagacaat tggaagagaa
     1861 aagagatatt taatcattat ttgaaccaac aaacgacttt tagtataacc acagaaattg
     1921 atattagtgt tttataccga aacataaaac aagaaggata taaattttac cctgcattta
     1981 ttttcttagt gacaagggtg ataaactcaa atacagcttt tagaactggt tacaatagcg
     2041 acggagagtt aggttattgg gataagttag agccacttta tacaattttt gatggtgtat
     2101 ctaaaacatt ctctggtatt tggactcctg taaagaatga cttcaaagag ttttatgatt
     2161 tatacctttc tgatgtagag aaatataatg gttcggggaa attgtttccc aaaacaccta
     2221 tacctgaaaa tgctttttct ctttctatta ttccatggac ttcatttact gggtttaact
     2281 taaatatcaa taataatagt aattaccttc tacccattat tacagcagga aaattcatta
     2341 ataaaggtaa ttcaatatat ttaccgctat ctttacaggt acatcattct gtttgtgatg
     2401 gttatcatgc aggattgttt atgaactcta ttcaggaatt gtcagatagg cctaatgact
     2461 ggcttttata atatgagata atgccgactg tactttttac agtcggtttt ctaatgtcac
     2521 taacctgccc cgttagttga agaaggtttt tatattacag ctccagatct atggtgcact
     2581 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     2641 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2701 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     2761 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2821 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2881 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2941 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3001 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3061 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3121 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3181 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3241 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3301 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3361 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3421 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3481 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3541 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3601 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3661 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3721 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3781 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3841 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3901 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3961 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4021 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4081 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4141 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4201 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4261 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4321 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4381 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4441 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4501 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4561 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4621 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4681 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4741 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     4801 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     4861 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     4921 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     4981 gattacgcca agcttgcatg cctgcaggtc atttattaac gattacctgc ttttaattct
     5041 gcatctaatt gcataagctc cgccttcatt ttttcctttt cagcttcaaa atcattattc
     5101 actggaagat cttctgctac tgcattttca tcatttgttt gtagccactt tggaaccatt
     5161 tctttacgtg cattacgtac tacacgtttc ctttttgttt gcttcggttt ccaatgttta
     5221 agacggtatt ctaatagtga tttatattgc ttattcgtat caacgattac agtatcgtta
     5281 agcatttcac ttttcacttc gtcgaaaggt acattattat atttttctcg taatttatca
     5341 tcaataaaat cacgtggact actactaatt ttttcatttg ttaatttttc atttgtttct
     5401 tttttattta taggtgtacg ttctgtgttg tacacactgt gttgtacaga acgtacatca
     5461 gaagaaattt gcttattttg tatctgatgt acattttgta catcagatat tttttcagtg
     5521 atttcaagtg gcgaatgttg tatcacgctt aacgtcatga cgtttttacc ttgcgattgt
     5581 aagagattta tttgttcaat gacaagcaat tcgaaattag cttgcgaaat cggaatatct
     5641 gatacggcgt aaaaataatc tttttttccg tcaacatatg cgctaaaacc gacagcgtat
     5701 tgtttaatag ctaattcctt ccaagcagcg tctacatttc tacgagaaaa tttgttttgt
     5761 aactgggtct tcctaatcgt ccaaccttca cctaagctca taatatgact taatagaccg
     5821 atagaccgca agtcctccag gtccgtttgt aatggcgcgt tatgtatttg agcatattgc
     5881 gttgttttta ttgttttgat aattccattc atttctaaat tcctccaacg attgattttt
     5941 tgttggagta cttgctataa tatacgtaac caaataaacg tactattgca acaagtacaa
     6001 ccaagagaac ggaggcctca atctccgttc ttttttatta tatcatgtcc tattgatgaa
     6061 aataggagtg ggtataaaat cggaatttta ctagagacat tgattaattt gcagacagtt
     6121 ttagttaaca gttgaatata aatgactcta gaggatcctt aggcaaatac gtattcagaa
     6181 tccatgattt cgttaaggtc gaactcccct tgctcaggta cttcgatacc tgtatcaacc
     6241 atccagcgct cttcatgctc ctccagtagt ttctgaagcg cattaccatc aatatacatt
     6301 gcaaccatct gccctttaag tgccactcta agagtgaggg tgttgtctgc atgagtacca
     6361 aaagagtcgt ggattacagc gatactagtt acgcccttgt ctaccaattc acatacggta
     6421 aggataaggt gacttgcgtc atgaccgtgt acgaaattag gtgctgctgc tcccatcata
     6481 gcggcttcat ctacgatatc cgtttcaacc tgaagggaca tcttgatatc acccatcaga
     6541 caggtacgca cgcgtagcat ctcggttgcc atgatcttct gttctaagat gaagccagta
     6601 ggcagggtgt acatcaggcc ttcattacgt ttcgctgcaa agcgtgcaag ctggcgtatc
     6661 atcttcatag ctactatcgg tgccttaact acttcagaaa tagaaggcca gattagtgcc
     6721 gtcatgtagt tgtaagctgc gcccggagtc aagtaatctt gacgatcgtc ttcaaaagga
     6781 tgtaccttgt ttgccgtccg cccttctgct actgccttct gcgcctcttt ttcctctaag
     6841 tctacgatgt aatcaatcac agattcacgg caagttaagc gagtagaacc atatggcaag
     6901 gtcatcacgg gctttttggt taagctacgg gtaataccaa tactatccca tgcgctagcc
     6961 attgctcgca gttctgtacc ggacagcgtg acgctaccag aagtaaacgt ggttgcatcg
     7021 tccgcatcca tatatagcgc attcttcttg ataaccactt gcgccaccgc cccatagata
     7081 tcctgcggtg catcggaggg tttcaggtta acagctttgg cccctacttc gtcgcgaagc
     7141 atagcactat agtgctgaat gcctgaacaa gacccgtcct gatgtactgg taggtgagtg
     7201 cggaattcgt cggcccttcc ttcatccacc aaatcaaggt attgagcata ctcaaagcac
     7261 caagcgagga attcataagg tgcatcagct ttagcccatt gggtgaatgt gagagggtct
     7321 gcggcgatgt ctcgacacat atcttggaat tcctcatcta atacgttaga cacgcgcaca
     7381 tcaaaagttt tcttgtccca tccccaaagg ttagcaccat tgatgcagaa ccatttaagc
     7441 gcttctacgc cattcacagg gcgtccctcg gtaaagcgga gtaatgcctt acctaagtcg
     7501 ttagactgcg gagagagcgt gctagattgc acatagacac ggctgcggct atccattgcg
     7561 tacacgaagt aaatggattc aaaggcgcta tatttacggg cctgtcctac catgcgaaca
     7621 acggcggcgg actttgaacc gcgcttagtt tctgcggtat ataggcgcgc gcattcgcct
     7681 ttccagttaa tgaattgttg ccactgctca ggtgatagca tctctttcag ttcacgaccg
     7741 cgcaggtgtt ggaattcaac aggtaccggg ttagctggct tgttctcctt gtcaatcagt
     7801 ggcttgaagg aaggtacacc ataaccaagg tctaagcgga ttgcttcttc aataactgct
     7861 aatacatcct tgttgatttg ccattgtgta ttttgtaatg cgttgatagc cttataaacc
     7921 tttggcattt gcttttgagt caacttgcgt acatgctcac ggttaccttt tacaagacgg
     7981 atacggctag ctaccttctc agtatggaac cctccattaa atggagttct ccaaggacga
     8041 ggagggatta cgcaaggggc ataagctggg cttaattgcg ctacgtgctc tttgaatgcg
     8101 ctaatccact ggcctacact ttcagaagtt tgtaagtagt aaatagtctt tccgccataa
     8161 gtgcgcatag cacgcataaa tacaggttca ccattataga aaacgctacc ttctaagatt
     8221 tcaagcaagg tagtaccaat ctgcaattga gtttcttttg gccacgcctc ccaacggtca
     8281 aagtccgcgt ccttttctgc aactgatttt tcagcaacta cagctacgtt atgagcgtga
     8341 cgatatgact tagtacggct agccttgagt gacttcttaa ccttctcaaa gtatttagcg
     8401 gcgtgacctt ctagcttaga aaagcgcact tggtcttcaa tgcgttctgc tacactcatt
     8461 gcaatagcct gaagggtagc atccgtattc agcatatcca taacaacttt catagtgatg
     8521 tatgctgcaa cttcattttc tacacattgt aagaaagcca atgcgcgagg tgcacgacct
     8581 ttcttacctt cgtactcttc tttataagcc tgaatgcctt cagccatagg tgcaataagt
     8641 tctgacaaca ggcggcggtt ccatgctgtg tcgctctcgc tacctgctgc aatctggcgt
     8701 tgttgatctg cttcgaagcg acgaatgcca ccattaaaca tctcttcttc taattgaagc
     8761 tggatagcgt gtaaatcttg agct
//
