LOCUS       sequence-69934-e        5234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector E. coli/B.meg., encoding a promoter and a terminator 
            for RNA polymerase of the phage K1E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5234)
  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 5234)
  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-2+t
FEATURES             Location/Qualifiers
     source          1..5234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(993..1997)
                     /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"
     rep_origin      complement(2763..3351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2843..2862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3522..4313,4314..4382))
                     /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     4145..4164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4383..4487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4555..4573
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(4651..5234,1..793))
                     /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
                     "
ORIGIN
        1 cacaaagaac tccaatcata aaaggtatat ttttccctaa tccgggatca acaaaaggat
       61 ctgttacttt cctgatatgt tttacaaata tcaggaatga cagcacgcta acgataagaa
      121 aagaaatgct atatgatgtt gtaaacaaca taaaaaatac aatgcctaca gacattagta
      181 taattccttt gatatcaaaa tgacctttta tccttacttc tttctttaat aatttcataa
      241 gaaacggaac agtgataatt gttatcatag gaatgagtag aagataggac caatgaatat
      301 aatgggctat cattccacca atcgctggac cgactccttc tcccatggct actatcgatc
      361 caataagacc aaatgcttta cccctatttt cctttggaat atagcgcgca actacaacca
      421 ttacgagtgc tggaaatgca gctgcaccag ccccttgaat aaaacgagcc ataataagta
      481 aggaaaagaa agaatggcca acaaacccaa ttaccgaccc gaaacaattt attataattc
      541 caaataggag taaccttttg atgcctaatt gatcagatag ctttccatat acagctgttc
      601 caatggaaaa ggttaacata aaggctgtgt tcacccagtt tgtactcgca ggtggtttat
      661 taaaatcatt tgcaatatca ggtaatgaga cgttcaaaac catttcattt aatacgctaa
      721 aaaaagataa aatgcaaagc caaattaaaa tttggttgtg tcgtaaattc gattgtgaat
      781 aggatgtatt cacatttcac cctccaataa tgagggcaga cgtagtttat agggttaatg
      841 atacgcttcc ctcttttaat tgaaccctgt tacattcatt acacttcata attaattcct
      901 cctaaacttg attaaaacat tttaccacat ataaactaag ttttaaattc agtatttcat
      961 cacttataca acaatatggc ccgtttgttg aactactctt taataaaata atttttccgt
     1021 tcccaattcc acattgcaat aatagaaaat ccatcttcat cggctttttc gtcatcatct
     1081 gtatgaatca aatcgccttc ttctgtgtca tcaaggttta attttttatg tatttctttt
     1141 aacaaaccac cataggagat taacctttta cggtgtaaac cttcctccaa atcagacaaa
     1201 cgtttcaaat tcttttcttc atcatcggtc ataaaatccg tatcctttac aggatatttt
     1261 gcagtttcgt caattgccga ttgtatatcc gatttatatt tatttttcgg tcgaatcatt
     1321 tgaactttta catttggatc atagtctaat ttcattgcct ttttccaaaa ttgaatccat
     1381 tgtttttgat tcacgtagtt ttctgtattc ttaaaataag ttggttccac acataccaat
     1441 acatgcatgt gctgattata agaattatct ttattattta ttgtcacttc cgttgcacgc
     1501 ataaaaccaa caagattttt attaattttt ttatattgca tcattcggcg aaatccttga
     1561 gccatatctg acaaactctt atttaattct tcgccatcat aaacattttt aactgttaat
     1621 gtgagaaaca accaacgaac tgttggcttt tgtttaataa cttcagcaac aaccttttgt
     1681 gactgaatgc catgtttcat tgctctcctc cagttgcaca ttggacaaag cctggattta
     1741 caaaaccaca ctcgatacaa ctttctttcg cctgtttcac gattttgttt atactctaat
     1801 atttcagcac aatcttttac tctttcagcc tttttaaatt caagaatatg cagaagttca
     1861 aagtaatcaa cattagcgat tttcttttct ctccatggtc tcacttttcc actttttgtc
     1921 ttgtccacta aaacccttga tttttcatct gaataaatgc tactattagg acacataata
     1981 ttaaaagaaa cccccatcta tttagttatt tgtttggtca cttataactt taacagatgg
     2041 ggtttttctg tgcaaccaat tttaagggtt ttcaatactt taaaacacat acataccaac
     2101 acttcaacgc acctttcagc aactaaaata aaaatgacgt tatttctata tgtatcaaga
     2161 taagaaagaa caagttcaaa accatcaaaa aaagacacct tttcaggtgc tttttttatt
     2221 ttataaactc attccctgat ctcgacttcg ttcttttttt acctctcggt tatgagttag
     2281 ttcaaattcg ttctttttag gttctaaatc gtgtttttct tggaattgtg ctgttttatc
     2341 ctttaccttg tctacaaacc ccttaaaaac gtttttaaag gcttttaagc cgtctgtacg
     2401 ttccttaagc aaactattta gctgacacta taagagaagg cttaacaagg cgttgctacg
     2461 gtagcgcctg attaaacttt cacttacaag gaggtgaaat gtacaatggt ccaaactagt
     2521 tcgaagatct ccggagctcc cgggatccgg taccggccgc atgctaacta gctaaccggt
     2581 tacctatgcc ctatctacct tgcgtaggta gggttctttt tgttgccggt ccaagaattg
     2641 gagctaatta attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat
     2701 ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggccgc gttgctggcg
     2761 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2881 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2941 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3001 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3061 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3181 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     3241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3361 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3421 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3661 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     3841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     4141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4381 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4441 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4501 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4561 cgtatcacga ggccctttcg tcttcaagaa ttcctgttat aaaaaaagga tcaattttga
     4621 actctctccc aaagttgatc ccttaacgat ttagaaatcc ctttgagaat gtttatatac
     4681 attcaaggta accagccaac taatgacaat gattcctgaa aaaagtaata acaaattact
     4741 atacagataa gttgactgat caacttccat aggtaacaac ctttgatcaa gtaagggtat
     4801 ggataataaa ccacctacaa ttgcaatacc tgttccctct gataaaaagc tggtaaagtt
     4861 aagcaaactc attccagcac cagcttcctg ctgtttcaag ctacttgaaa caattgttga
     4921 tataactgtt ttggtgaacg aaagcccacc taaaacaaat acgattataa ttgtcatgaa
     4981 ccatgatgtt gtttctaaaa gaaaggaagc agttaaaaag ctaacagaaa gaaatgtaac
     5041 tccgatgttt aacacgtata aaggacctct tctatcaaca agtatcccac caatgtagcc
     5101 gaaaataatg acactcattg ttccagggaa aataattaca cttccgattt cggcagtact
     5161 tagctggtga acatctttca tcatataagg aaccatagag acaaaccctg ctactgttcc
     5221 aaatataatt cccc
//
