LOCUS       sequence-69927-e        8778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector E. coli/B.meg.; encoding the RNA polymerase of the 
            phage K1E under control of the xylose inducible promoter PxylA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8778)
  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 8778)
  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: pK1E-RNAP
FEATURES             Location/Qualifiers
     source          1..8778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3821..3837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3821..3837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3834..3856)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3845..3861
                     /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)."
     promoter        complement(join(3869..3875,3876..3893,3894..3899))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3914..3935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4052..4069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4223..4811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4303..4322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4982..5773,5774..5842))
                     /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     5605..5624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5843..5947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6015..6033
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6071..6093)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6193..6212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctagtcaaga cctacacgct attcaacttc aacttgaaga agaaatgttt aacggcggca
       61 tccgtcgctt tgaagcggac caacaacgcc agattgcatc cggtaatgaa tcagacacgg
      121 catggaatcg ccgcttattg tccgaactaa tcgcacctat ggcggaaggt attcaggctt
      181 acaaggaaga atacgaaggc aagagaggtc gtgcaccacg tgcattagct ttcattaact
      241 gcgtaggaaa cgaagtggca gcatatatca ccatgaaaat cgttatggat atgctgaata
      301 cagacgttac cttacaagct atcgctatga atgtagcaga ccgcatcgag gaccaagtac
      361 gttttagcaa gctggaaggt cacgcagcta agtactttga gaaagtgaag aagtcgctta
      421 aggctagcaa gactaaatct tatcgtcatg cgcacaacgt agcggtagta gctgagaaat
      481 ctgttgctga ccgtgacgcg gatttctccc gttgggaggc atggcctaaa gacaccttgc
      541 tgcaaatcgg tatgaccttg ctcgaaatat tggagaatag tgtattcttc aatggtcaac
      601 ctgtcttcct tcgtaccttg cgcactaacg gcggaaagca tggtgtttac tatttacaga
      661 ccagtgaaca tgttggcgag tggataactg catttaagga acatgtagcg cagctcagcc
      721 ctgcctatgc accttgcgtt atacctcccc gcccttgggt atcacctttt aacggtgggt
      781 ttcatactga gaaagtagca agccgtattc gtctggtaaa aggcaatcgt gagcacgtcc
      841 gcaagctgac caaaaagcaa atgccagccg tttataaggc tgttaacgct ttgcaggcaa
      901 ctaagtggca ggttaataaa gaggttttac aggttgtgga ggacgttata cgtctagacc
      961 tcggctatgg tgtaccttcc tttaaaccac tcatcgaccg tgagaacaag ccagctaatc
     1021 cggtgccgtt agagttccag cacctgcgag gccgtgaact gaaagaaatg ttaacaccgg
     1081 aacaatggca agccttcatc aactggaaag gtgaatgcac caagctgtat accgctgaga
     1141 ctaagcgcgg cagcaaatcg gcggcgaccg tccgcatggt agggcaggcc cgtaaataca
     1201 gccaatttga cgcaatatac ttcgtgtatg ctctggacag ccgcagccgc gtctacgcgc
     1261 aatctagcac gctctctccg caatcaaacg acttaggcaa ggcattgctc cgttttaccg
     1321 aagggcagcg tcttgatagc gctgaggcgc ttaagtggtt tttggtgaac ggggctaata
     1381 actggggttg ggataagaaa acttttgacg tgcgcaccgc taacgtgctg gatagtgaat
     1441 ttcaagacat gtgccgcgac attgcagcgg atccgctgac cttcactcaa tgggtaaatg
     1501 ccgactcccc ttacggcttc cttgcatggt gctttgaata tgcgcgttat ctggatgcac
     1561 tggatgaagg cacgcaagac caattcatga cgcacctccc agtccatcaa gatggtagtt
     1621 gttctggtat ccagcactac agtgctatgc tacgcgatgc agtaggtgcg aaagcagtaa
     1681 accttaagcc ctctgactct cctcaagata tttatggtgc cgttgcgcag gtagtaattc
     1741 agaagaatta tgcatacatg aatgcagagg atgcggaaac cttcacttct ggcagcgtga
     1801 ctttaacagg tgcggaactg cgtagtatgg ctagtgcgtg ggatatgata ggaatcactc
     1861 gcggcctgac caaaaagccc gtaatgacac taccttatgg cagcacacgt ctaacctgcc
     1921 gtgagtcagt gattgattat atcgttgatt tagaagaaaa agaggcccaa cgggctattg
     1981 cggaagggcg taccgccaat cctgtacacc cttttgataa tgaccgtaaa gacagcctga
     2041 cacctagcgc agcttataac tatatgacag ctttaatctg gccttctatc tcggaagtgg
     2101 ttaaagcccc tatagtggca atgaaaatga ttcgtcagct tgcacgtttc gcagctaaaa
     2161 gaaacgaagg cttagaatat cccttaccaa ctggcttcat cttgcaacaa aagataatgg
     2221 ctaccgatat gctccgcgta tccacttgct tgatgggtga aatcaagatg agtctccaga
     2281 ttgaaacaga tgtagttgat gaaacggcaa tgatgggtgc ggctgctcct aacttcgtcc
     2341 atggtcacga tgctagccac ctgattctga ctgtatgtga cctagtggat aaaggtatta
     2401 catccgttgc agtcatccat gattctttcg gtactcatgc aggacgtacc gcagacctgc
     2461 gggatagctt aagggaagaa atggttaaga tgtatcaaaa ccataatgcc ctgcaaaacc
     2521 tgctagatgt gcacgaagag cgttggttag tagacaccgg aatccaagta ccagagcaag
     2581 gggagtttga ccttaacgaa atcttagttt cagactattg tttcgcataa gagctcccgg
     2641 gatcctctag agtcatttat attcaactgt taactaaaac tgtctgcaaa ttaatcaatg
     2701 tctctagtaa aattccgatt ttatacccac tcctattttc atcaatagga catgatataa
     2761 taaaaaagaa cggagattga ggcctccgtt ctcttggttg tacttgttgc aatagtacgt
     2821 ttatttggtt acgtatatta tagcaagtac tccaacaaaa aatcaatcgt tggaggaatt
     2881 tagaaatgaa tggaattatc aaaacaataa aaacaacgca atatgctcaa atacataacg
     2941 cgccattaca aacggacctg gaggacttgc ggtctatcgg tctattaagt catattatga
     3001 gcttaggtga aggttggacg attaggaaga cccagttaca aaacaaattt tctcgtagaa
     3061 atgtagacgc tgcttggaag gaattagcta ttaaacaata cgctgtcggt tttagcgcat
     3121 atgttgacgg aaaaaaagat tatttttacg ccgtatcaga tattccgatt tcgcaagcta
     3181 atttcgaatt gcttgtcatt gaacaaataa atctcttaca atcgcaaggt aaaaacgtca
     3241 tgacgttaag cgtgatacaa cattcgccac ttgaaatcac tgaaaaaata tctgatgtac
     3301 aaaatgtaca tcagatacaa aataagcaaa tttcttctga tgtacgttct gtacaacaca
     3361 gtgtgtacaa cacagaacgt acacctataa ataaaaaaga aacaaatgaa aaattaacaa
     3421 atgaaaaaat tagtagtagt ccacgtgatt ttattgatga taaattacga gaaaaatata
     3481 ataatgtacc tttcgacgaa gtgaaaagtg aaatgcttaa cgatactgta atcgttgata
     3541 cgaataagca atataaatca ctattagaat accgtcttaa acattggaaa ccgaagcaaa
     3601 caaaaaggaa acgtgtagta cgtaatgcac gtaaagaaat ggttccaaag tggctacaaa
     3661 caaatgatga aaatgcagta gcagaagatc ttccagtgaa taatgatttt gaagctgaaa
     3721 aggaaaaaat gaaggcggag cttatgcaat tagatgcaga attaaaagca ggtaatcgtt
     3781 aataaatgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3841 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3901 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3961 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4021 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4081 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4621 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4981 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5041 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5101 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5161 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5281 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5341 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5401 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5521 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5581 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5641 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5701 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5761 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5821 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5881 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5941 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6001 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     6061 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     6121 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     6181 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataga tctggagctg
     6241 taatataaaa accttcttca actaacgggg caggttagtg acattagaaa accgactgta
     6301 aaaagtacag tcggcattat ctcatattat aaaagccagt cattaggcct atctgacaat
     6361 tcctgaatag agttcataaa caatcctgca tgataaccat cacaaacaga atgatgtacc
     6421 tgtaaagata gcggtaaata tattgaatta cctttattaa tgaattttcc tgctgtaata
     6481 atgggtagaa ggtaattact attattattg atatttaagt taaacccagt aaatgaagtc
     6541 catggaataa tagaaagaga aaaagcattt tcaggtatag gtgttttggg aaacaatttc
     6601 cccgaaccat tatatttctc tacatcagaa aggtataaat cataaaactc tttgaagtca
     6661 ttctttacag gagtccaaat accagagaat gttttagata caccatcaaa aattgtataa
     6721 agtggctcta acttatccca ataacctaac tctccgtcgc tattgtaacc agttctaaaa
     6781 gctgtatttg agtttatcac ccttgtcact aagaaaataa atgcagggta aaatttatat
     6841 ccttcttgtt ttatgtttcg gtataaaaca ctaatatcaa tttctgtggt tatactaaaa
     6901 gtcgtttgtt ggttcaaata atgattaaat atctcttttc tcttccaatt gtctaaatca
     6961 attttattaa agttcatttg atatgcctcc taaattttta tctaaagtga atttaggagg
     7021 cttacttgtc tgctttcttc attagaatca atcctttttt aaaagtcaat attactgtaa
     7081 cataaatata tattttaaaa atatcccact ttatccaatt ttcgtttgtt gaactaatgg
     7141 gtgctttagt tgaagaataa aagaccacat taaaaaatgt ggtcttttgt gtttttttaa
     7201 aggatttgag cgtagcgaaa aatccttttc tttcttatct tgataataag ggtaactatt
     7261 gccggatcgt cctcaggagt aggcgacatc gctaaataat gatctatgcg gtgtgaaata
     7321 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc ctttgcgttc acttaactaa
     7381 cttatagggg taacacttaa aaaagaatca ataacgatag aaaccgctcc taaagcaggt
     7441 gcattttttc ctaacgaaga aggcaatagt tcacatttat tgtctaaatg agaatggact
     7501 ctagaagaaa cttcgttttt aatcgtattt aaaacaatgg gatgagattc aattatatga
     7561 tttctcaaga taacagcttc tatatcaaat gtattaagga tattggttaa tccaattccg
     7621 atataaaagc caaagttttg aagtgcattt aacatttcta catcattttt atttgcgcgt
     7681 tccacaatct cttttcgaga aatattcttt tcttctttag agagcgaagc cagtaacgct
     7741 ttttcagaag catataattc ccaacagcct cgatttccac agctgcattt gggtccatta
     7801 aaatctatcg tcatatgacc catttcccca gaaaaaccct gaacaccttt atacaattcg
     7861 ttgttaataa caagtccagt tccaattccg atattaatac tgatgtaaac gatgttttca
     7921 tagttttttg tcataccaaa tactttttca ccgtatgctc ctgcattagc ttcattttca
     7981 acaaaaaccg gaacattaaa ctcactctca attaaaaact gcaaatcttt gatattccaa
     8041 tttaagttag gcatgaaaat aatttgctga tgacgatcta caaggcctgg aacacaaatt
     8101 cctattccga ctagaccata aggggactca ggcatatggg ttacaaaacc atgaataagt
     8161 gcaaataaaa tctcttttac ttcactagcg gaagaactag acaagtcaga agtcttctcg
     8221 agaataatat ttccttctaa gtcggttaga attccgttaa gatagtcgac tcctatatca
     8281 ataccaatcg agtagcctgc attcttatta aaaacaagca ttacaggtct tctgccgcct
     8341 ctagattgcc ctgccccaat ttcaaaaata aaatcttttt caagcagtgt atttacttga
     8401 gaggagacag tagacttgtt taatcctgta atctcagaga gagttgccct ggagacaggg
     8461 gagttcttca aaatttcatc taatattaat ttttgattca ttttttttac taaagcttga
     8521 tctgcaattt gaataataac cactcctttg tttatccacc gaactaagtt ggtgtttttt
     8581 gaagcttgaa ttagatattt aaaagtatca tatctaatat tataactaaa ttttctaaaa
     8641 aaaacattga aataaacatt tattttgtat atgatgagat aaagttagtt tattggataa
     8701 acaaactaac tcaattaaga tagttgatgg ataaacttgt tcacttaaat caaaggggga
     8761 aatgacaaat ggtccaaa
//
