LOCUS       sequence-69929-e        5183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector E. coli/B.meg., encoding a promoter for RNA 
            polymerase of the phage K1E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5183)
  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 5183)
  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: pPK1E-1
FEATURES             Location/Qualifiers
     source          1..5183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             795..806
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     RBS             837..845
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     rep_origin      complement(1077..1665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1157..1176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1836..2627,2628..2696))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2459..2478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2697..2801)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2869..2887
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(2965..4341)
                     /codon_start=1
                     /gene="Bacillus subtilis tetL"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     CDS             complement(join(4541..5183,1..362))
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
ORIGIN
        1 cgaactgttg gcttttgttt aataacttca gcaacaacct tttgtgactg aatgccatgt
       61 ttcattgctc tcctccagtt gcacattgga caaagcctgg atttacaaaa ccacactcga
      121 tacaactttc tttcgcctgt ttcacgattt tgtttatact ctaatatttc agcacaatct
      181 tttactcttt cagccttttt aaattcaaga atatgcagaa gttcaaagta atcaacatta
      241 gcgattttct tttctctcca tggtctcact tttccacttt ttgtcttgtc cactaaaacc
      301 cttgattttt catctgaata aatgctacta ttaggacaca taatattaaa agaaaccccc
      361 atctatttag ttatttgttt ggtcacttat aactttaaca gatggggttt ttctgtgcaa
      421 ccaattttaa gggttttcaa tactttaaaa cacatacata ccaacacttc aacgcacctt
      481 tcagcaacta aaataaaaat gacgttattt ctatatgtat caagataaga aagaacaagt
      541 tcaaaaccat caaaaaaaga caccttttca ggtgcttttt ttattttata aactcattcc
      601 ctgatctcga cttcgttctt tttttacctc tcggttatga gttagttcaa attcgttctt
      661 tttaggttct aaatcgtgtt tttcttggaa ttgtgctgtt ttatccttta ccttgtctac
      721 aaacccctta aaaacgtttt taaaggcttt taagccgtct gtacgttcct taagatcagt
      781 atttactgga cactatagaa gggagaaatg tctccctaaa ttatcaattt gatttataag
      841 gaggtgaaat gtacaatggt ccaaactagt tcgaagatct ccggagctcc cgggatccgg
      901 taccggccgc atgccggcta aacctcgcga acggattcac cggtccaaga attggagcta
      961 attaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg
     1021 cgtccgccat ctccagcagc cgcacgcggc gcatctcggg ccgcgttgct ggcgtttttc
     1081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1501 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1801 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1861 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1921 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1981 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2041 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2101 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     2161 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2221 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2281 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     2341 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     2401 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     2461 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2521 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2581 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2641 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2701 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2761 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2821 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2881 acgaggccct ttcgtcttca agaattcctg ttataaaaaa aggatcaatt ttgaactctc
     2941 tcccaaagtt gatcccttaa cgatttagaa atccctttga gaatgtttat atacattcaa
     3001 ggtaaccagc caactaatga caatgattcc tgaaaaaagt aataacaaat tactatacag
     3061 ataagttgac tgatcaactt ccataggtaa caacctttga tcaagtaagg gtatggataa
     3121 taaaccacct acaattgcaa tacctgttcc ctctgataaa aagctggtaa agttaagcaa
     3181 actcattcca gcaccagctt cctgctgttt caagctactt gaaacaattg ttgatataac
     3241 tgttttggtg aacgaaagcc cacctaaaac aaatacgatt ataattgtca tgaaccatga
     3301 tgttgtttct aaaagaaagg aagcagttaa aaagctaaca gaaagaaatg taactccgat
     3361 gtttaacacg tataaaggac ctcttctatc aacaagtatc ccaccaatgt agccgaaaat
     3421 aatgacactc attgttccag ggaaaataat tacacttccg atttcggcag tacttagctg
     3481 gtgaacatct ttcatcatat aaggaaccat agagacaaac cctgctactg ttccaaatat
     3541 aattccccca caaagaactc caatcataaa aggtatattt ttccctaatc cgggatcaac
     3601 aaaaggatct gttactttcc tgatatgttt tacaaatatc aggaatgaca gcacgctaac
     3661 gataagaaaa gaaatgctat atgatgttgt aaacaacata aaaaatacaa tgcctacaga
     3721 cattagtata attcctttga tatcaaaatg accttttatc cttacttctt tctttaataa
     3781 tttcataaga aacggaacag tgataattgt tatcatagga atgagtagaa gataggacca
     3841 atgaatataa tgggctatca ttccaccaat cgctggaccg actccttctc ccatggctac
     3901 tatcgatcca ataagaccaa atgctttacc cctattttcc tttggaatat agcgcgcaac
     3961 tacaaccatt acgagtgctg gaaatgcagc tgcaccagcc ccttgaataa aacgagccat
     4021 aataagtaag gaaaagaaag aatggccaac aaacccaatt accgacccga aacaatttat
     4081 tataattcca aataggagta accttttgat gcctaattga tcagatagct ttccatatac
     4141 agctgttcca atggaaaagg ttaacataaa ggctgtgttc acccagtttg tactcgcagg
     4201 tggtttatta aaatcatttg caatatcagg taatgagacg ttcaaaacca tttcatttaa
     4261 tacgctaaaa aaagataaaa tgcaaagcca aattaaaatt tggttgtgtc gtaaattcga
     4321 ttgtgaatag gatgtattca catttcaccc tccaataatg agggcagacg tagtttatag
     4381 ggttaatgat acgcttccct cttttaattg aaccctgtta cattcattac acttcataat
     4441 taattcctcc taaacttgat taaaacattt taccacatat aaactaagtt ttaaattcag
     4501 tatttcatca cttatacaac aatatggccc gtttgttgaa ctactcttta ataaaataat
     4561 ttttccgttc ccaattccac attgcaataa tagaaaatcc atcttcatcg gctttttcgt
     4621 catcatctgt atgaatcaaa tcgccttctt ctgtgtcatc aaggtttaat tttttatgta
     4681 tttcttttaa caaaccacca taggagatta accttttacg gtgtaaacct tcctccaaat
     4741 cagacaaacg tttcaaattc ttttcttcat catcggtcat aaaatccgta tcctttacag
     4801 gatattttgc agtttcgtca attgccgatt gtatatccga tttatattta tttttcggtc
     4861 gaatcatttg aacttttaca tttggatcat agtctaattt cattgccttt ttccaaaatt
     4921 gaatccattg tttttgattc acgtagtttt ctgtattctt aaaataagtt ggttccacac
     4981 ataccaatac atgcatgtgc tgattataag aattatcttt attatttatt gtcacttccg
     5041 ttgcacgcat aaaaccaaca agatttttat taattttttt atattgcatc attcggcgaa
     5101 atccttgagc catatctgac aaactcttat ttaattcttc gccatcataa acatttttaa
     5161 ctgttaatgt gagaaacaac caa
//
