LOCUS       sequence-69932-e        5198 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 5198)
  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 5198)
  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
FEATURES             Location/Qualifiers
     source          1..5198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1147..1735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1227..1246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1906..2697,2698..2766))
                     /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     2529..2548
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2767..2871)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2939..2957
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(3035..4411)
                     /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(4611..5198,1..417))
                     /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 atttaattct tcgccatcat aaacattttt aactgttaat gtgagaaaca accaacgaac
       61 tgttggcttt tgtttaataa cttcagcaac aaccttttgt gactgaatgc catgtttcat
      121 tgctctcctc cagttgcaca ttggacaaag cctggattta caaaaccaca ctcgatacaa
      181 ctttctttcg cctgtttcac gattttgttt atactctaat atttcagcac aatcttttac
      241 tctttcagcc tttttaaatt caagaatatg cagaagttca aagtaatcaa cattagcgat
      301 tttcttttct ctccatggtc tcacttttcc actttttgtc ttgtccacta aaacccttga
      361 tttttcatct gaataaatgc tactattagg acacataata ttaaaagaaa cccccatcta
      421 tttagttatt tgtttggtca cttataactt taacagatgg ggtttttctg tgcaaccaat
      481 tttaagggtt ttcaatactt taaaacacat acataccaac acttcaacgc acctttcagc
      541 aactaaaata aaaatgacgt tatttctata tgtatcaaga taagaaagaa caagttcaaa
      601 accatcaaaa aaagacacct tttcaggtgc tttttttatt ttataaactc attccctgat
      661 ctcgacttcg ttcttttttt acctctcggt tatgagttag ttcaaattcg ttctttttag
      721 gttctaaatc gtgtttttct tggaattgtg ctgttttatc ctttaccttg tctacaaacc
      781 ccttaaaaac gtttttaaag gcttttaagc cgtctgtacg ttccttaagc aaactattta
      841 gctgacacta taagagaagg cttaacaagg cgttgctacg gtagcgcctg attaaacttt
      901 cacttacaag gaggtgaaat gtacaatggt ccaaactagt tcgaagatct ccggagctcc
      961 cgggatccgg taccggccgc atgccggcta aacctcgcga acggattcac cggtccaaga
     1021 attggagcta attaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
     1081 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg ccgcgttgct
     1141 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1201 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1261 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1321 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1381 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1441 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1501 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1561 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     1621 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1681 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1741 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1801 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1861 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1921 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1981 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2041 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2101 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2161 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2221 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2281 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2341 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2401 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2461 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2521 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2581 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2641 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2701 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2761 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2821 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2881 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     2941 taggcgtatc acgaggccct ttcgtcttca agaattcctg ttataaaaaa aggatcaatt
     3001 ttgaactctc tcccaaagtt gatcccttaa cgatttagaa atccctttga gaatgtttat
     3061 atacattcaa ggtaaccagc caactaatga caatgattcc tgaaaaaagt aataacaaat
     3121 tactatacag ataagttgac tgatcaactt ccataggtaa caacctttga tcaagtaagg
     3181 gtatggataa taaaccacct acaattgcaa tacctgttcc ctctgataaa aagctggtaa
     3241 agttaagcaa actcattcca gcaccagctt cctgctgttt caagctactt gaaacaattg
     3301 ttgatataac tgttttggtg aacgaaagcc cacctaaaac aaatacgatt ataattgtca
     3361 tgaaccatga tgttgtttct aaaagaaagg aagcagttaa aaagctaaca gaaagaaatg
     3421 taactccgat gtttaacacg tataaaggac ctcttctatc aacaagtatc ccaccaatgt
     3481 agccgaaaat aatgacactc attgttccag ggaaaataat tacacttccg atttcggcag
     3541 tacttagctg gtgaacatct ttcatcatat aaggaaccat agagacaaac cctgctactg
     3601 ttccaaatat aattccccca caaagaactc caatcataaa aggtatattt ttccctaatc
     3661 cgggatcaac aaaaggatct gttactttcc tgatatgttt tacaaatatc aggaatgaca
     3721 gcacgctaac gataagaaaa gaaatgctat atgatgttgt aaacaacata aaaaatacaa
     3781 tgcctacaga cattagtata attcctttga tatcaaaatg accttttatc cttacttctt
     3841 tctttaataa tttcataaga aacggaacag tgataattgt tatcatagga atgagtagaa
     3901 gataggacca atgaatataa tgggctatca ttccaccaat cgctggaccg actccttctc
     3961 ccatggctac tatcgatcca ataagaccaa atgctttacc cctattttcc tttggaatat
     4021 agcgcgcaac tacaaccatt acgagtgctg gaaatgcagc tgcaccagcc ccttgaataa
     4081 aacgagccat aataagtaag gaaaagaaag aatggccaac aaacccaatt accgacccga
     4141 aacaatttat tataattcca aataggagta accttttgat gcctaattga tcagatagct
     4201 ttccatatac agctgttcca atggaaaagg ttaacataaa ggctgtgttc acccagtttg
     4261 tactcgcagg tggtttatta aaatcatttg caatatcagg taatgagacg ttcaaaacca
     4321 tttcatttaa tacgctaaaa aaagataaaa tgcaaagcca aattaaaatt tggttgtgtc
     4381 gtaaattcga ttgtgaatag gatgtattca catttcaccc tccaataatg agggcagacg
     4441 tagtttatag ggttaatgat acgcttccct cttttaattg aaccctgtta cattcattac
     4501 acttcataat taattcctcc taaacttgat taaaacattt taccacatat aaactaagtt
     4561 ttaaattcag tatttcatca cttatacaac aatatggccc gtttgttgaa ctactcttta
     4621 ataaaataat ttttccgttc ccaattccac attgcaataa tagaaaatcc atcttcatcg
     4681 gctttttcgt catcatctgt atgaatcaaa tcgccttctt ctgtgtcatc aaggtttaat
     4741 tttttatgta tttcttttaa caaaccacca taggagatta accttttacg gtgtaaacct
     4801 tcctccaaat cagacaaacg tttcaaattc ttttcttcat catcggtcat aaaatccgta
     4861 tcctttacag gatattttgc agtttcgtca attgccgatt gtatatccga tttatattta
     4921 tttttcggtc gaatcatttg aacttttaca tttggatcat agtctaattt cattgccttt
     4981 ttccaaaatt gaatccattg tttttgattc acgtagtttt ctgtattctt aaaataagtt
     5041 ggttccacac ataccaatac atgcatgtgc tgattataag aattatcttt attatttatt
     5101 gtcacttccg ttgcacgcat aaaaccaaca agatttttat taattttttt atattgcatc
     5161 attcggcgaa atccttgagc catatctgac aaactctt
//
