LOCUS       sequence-69933-e        5219 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 5219)
  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 5219)
  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+t
FEATURES             Location/Qualifiers
     source          1..5219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1024..2028)
                     /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"
     CDS             2461..2472
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     RBS             2503..2511
                     /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(2779..3367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2859..2878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3538..4329,4330..4398))
                     /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     4161..4180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4399..4503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4571..4589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(4667..5219,1..824))
                     /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 aaccctgcta ctgttccaaa tataattccc ccacaaagaa ctccaatcat aaaaggtata
       61 tttttcccta atccgggatc aacaaaagga tctgttactt tcctgatatg ttttacaaat
      121 atcaggaatg acagcacgct aacgataaga aaagaaatgc tatatgatgt tgtaaacaac
      181 ataaaaaata caatgcctac agacattagt ataattcctt tgatatcaaa atgacctttt
      241 atccttactt ctttctttaa taatttcata agaaacggaa cagtgataat tgttatcata
      301 ggaatgagta gaagatagga ccaatgaata taatgggcta tcattccacc aatcgctgga
      361 ccgactcctt ctcccatggc tactatcgat ccaataagac caaatgcttt acccctattt
      421 tcctttggaa tatagcgcgc aactacaacc attacgagtg ctggaaatgc agctgcacca
      481 gccccttgaa taaaacgagc cataataagt aaggaaaaga aagaatggcc aacaaaccca
      541 attaccgacc cgaaacaatt tattataatt ccaaatagga gtaacctttt gatgcctaat
      601 tgatcagata gctttccata tacagctgtt ccaatggaaa aggttaacat aaaggctgtg
      661 ttcacccagt ttgtactcgc aggtggttta ttaaaatcat ttgcaatatc aggtaatgag
      721 acgttcaaaa ccatttcatt taatacgcta aaaaaagata aaatgcaaag ccaaattaaa
      781 atttggttgt gtcgtaaatt cgattgtgaa taggatgtat tcacatttca ccctccaata
      841 atgagggcag acgtagttta tagggttaat gatacgcttc cctcttttaa ttgaaccctg
      901 ttacattcat tacacttcat aattaattcc tcctaaactt gattaaaaca ttttaccaca
      961 tataaactaa gttttaaatt cagtatttca tcacttatac aacaatatgg cccgtttgtt
     1021 gaactactct ttaataaaat aatttttccg ttcccaattc cacattgcaa taatagaaaa
     1081 tccatcttca tcggcttttt cgtcatcatc tgtatgaatc aaatcgcctt cttctgtgtc
     1141 atcaaggttt aattttttat gtatttcttt taacaaacca ccataggaga ttaacctttt
     1201 acggtgtaaa ccttcctcca aatcagacaa acgtttcaaa ttcttttctt catcatcggt
     1261 cataaaatcc gtatccttta caggatattt tgcagtttcg tcaattgccg attgtatatc
     1321 cgatttatat ttatttttcg gtcgaatcat ttgaactttt acatttggat catagtctaa
     1381 tttcattgcc tttttccaaa attgaatcca ttgtttttga ttcacgtagt tttctgtatt
     1441 cttaaaataa gttggttcca cacataccaa tacatgcatg tgctgattat aagaattatc
     1501 tttattattt attgtcactt ccgttgcacg cataaaacca acaagatttt tattaatttt
     1561 tttatattgc atcattcggc gaaatccttg agccatatct gacaaactct tatttaattc
     1621 ttcgccatca taaacatttt taactgttaa tgtgagaaac aaccaacgaa ctgttggctt
     1681 ttgtttaata acttcagcaa caaccttttg tgactgaatg ccatgtttca ttgctctcct
     1741 ccagttgcac attggacaaa gcctggattt acaaaaccac actcgataca actttctttc
     1801 gcctgtttca cgattttgtt tatactctaa tatttcagca caatctttta ctctttcagc
     1861 ctttttaaat tcaagaatat gcagaagttc aaagtaatca acattagcga ttttcttttc
     1921 tctccatggt ctcacttttc cactttttgt cttgtccact aaaacccttg atttttcatc
     1981 tgaataaatg ctactattag gacacataat attaaaagaa acccccatct atttagttat
     2041 ttgtttggtc acttataact ttaacagatg gggtttttct gtgcaaccaa ttttaagggt
     2101 tttcaatact ttaaaacaca tacataccaa cacttcaacg cacctttcag caactaaaat
     2161 aaaaatgacg ttatttctat atgtatcaag ataagaaaga acaagttcaa aaccatcaaa
     2221 aaaagacacc ttttcaggtg ctttttttat tttataaact cattccctga tctcgacttc
     2281 gttctttttt tacctctcgg ttatgagtta gttcaaattc gttcttttta ggttctaaat
     2341 cgtgtttttc ttggaattgt gctgttttat cctttacctt gtctacaaac cccttaaaaa
     2401 cgtttttaaa ggcttttaag ccgtctgtac gttccttaag atcagtattt actggacact
     2461 atagaaggga gaaatgtctc cctaaattat caatttgatt tataaggagg tgaaatgtac
     2521 aatggtccaa actagttcga agatctccgg agctcccggg atccggtacc ggccgcatgc
     2581 taactagcta accggttacc tatgccctat ctaccttgcg taggtagggt tctttttgtt
     2641 gccggtccaa gaattggagc taattaattc ttgcggagaa ctgtgaatgc gcaaaccaac
     2701 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg
     2761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3181 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3421 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3481 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3661 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3721 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3781 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3841 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3901 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3961 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4021 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4081 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4141 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4201 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4261 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4321 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4381 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4441 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4501 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4561 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattcc tgttataaaa
     4621 aaaggatcaa ttttgaactc tctcccaaag ttgatccctt aacgatttag aaatcccttt
     4681 gagaatgttt atatacattc aaggtaacca gccaactaat gacaatgatt cctgaaaaaa
     4741 gtaataacaa attactatac agataagttg actgatcaac ttccataggt aacaaccttt
     4801 gatcaagtaa gggtatggat aataaaccac ctacaattgc aatacctgtt ccctctgata
     4861 aaaagctggt aaagttaagc aaactcattc cagcaccagc ttcctgctgt ttcaagctac
     4921 ttgaaacaat tgttgatata actgttttgg tgaacgaaag cccacctaaa acaaatacga
     4981 ttataattgt catgaaccat gatgttgttt ctaaaagaaa ggaagcagtt aaaaagctaa
     5041 cagaaagaaa tgtaactccg atgtttaaca cgtataaagg acctcttcta tcaacaagta
     5101 tcccaccaat gtagccgaaa ataatgacac tcattgttcc agggaaaata attacacttc
     5161 cgatttcggc agtacttagc tggtgaacat ctttcatcat ataaggaacc atagagaca
//
