LOCUS       sequence-69936-e        5252 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 SP6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5252)
  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 5252)
  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: pPSP6+t
FEATURES             Location/Qualifiers
     source          1..5252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1101..2105)
                     /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"
     promoter        2524..2542
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2524..2541
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     rep_origin      complement(2889..3477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2969..2988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3648..4439,4440..4508))
                     /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     4271..4290
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4509..4613)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4681..4699
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(join(4777..5252,1..901))
                     /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 ggaaaataat tacacttccg atttcggcag tacttagctg gtgaacatct ttcatcatat
       61 aaggaaccat agagacaaac cctgctactg ttccaaatat aattccccca caaagaactc
      121 caatcataaa aggtatattt ttccctaatc cgggatcaac aaaaggatct gttactttcc
      181 tgatatgttt tacaaatatc aggaatgaca gcacgctaac gataagaaaa gaaatgctat
      241 atgatgttgt aaacaacata aaaaatacaa tgcctacaga cattagtata attcctttga
      301 tatcaaaatg accttttatc cttacttctt tctttaataa tttcataaga aacggaacag
      361 tgataattgt tatcatagga atgagtagaa gataggacca atgaatataa tgggctatca
      421 ttccaccaat cgctggaccg actccttctc ccatggctac tatcgatcca ataagaccaa
      481 atgctttacc cctattttcc tttggaatat agcgcgcaac tacaaccatt acgagtgctg
      541 gaaatgcagc tgcaccagcc ccttgaataa aacgagccat aataagtaag gaaaagaaag
      601 aatggccaac aaacccaatt accgacccga aacaatttat tataattcca aataggagta
      661 accttttgat gcctaattga tcagatagct ttccatatac agctgttcca atggaaaagg
      721 ttaacataaa ggctgtgttc acccagtttg tactcgcagg tggtttatta aaatcatttg
      781 caatatcagg taatgagacg ttcaaaacca tttcatttaa tacgctaaaa aaagataaaa
      841 tgcaaagcca aattaaaatt tggttgtgtc gtaaattcga ttgtgaatag gatgtattca
      901 catttcaccc tccaataatg agggcagacg tagtttatag ggttaatgat acgcttccct
      961 cttttaattg aaccctgtta cattcattac acttcataat taattcctcc taaacttgat
     1021 taaaacattt taccacatat aaactaagtt ttaaattcag tatttcatca cttatacaac
     1081 aatatggccc gtttgttgaa ctactcttta ataaaataat ttttccgttc ccaattccac
     1141 attgcaataa tagaaaatcc atcttcatcg gctttttcgt catcatctgt atgaatcaaa
     1201 tcgccttctt ctgtgtcatc aaggtttaat tttttatgta tttcttttaa caaaccacca
     1261 taggagatta accttttacg gtgtaaacct tcctccaaat cagacaaacg tttcaaattc
     1321 ttttcttcat catcggtcat aaaatccgta tcctttacag gatattttgc agtttcgtca
     1381 attgccgatt gtatatccga tttatattta tttttcggtc gaatcatttg aacttttaca
     1441 tttggatcat agtctaattt cattgccttt ttccaaaatt gaatccattg tttttgattc
     1501 acgtagtttt ctgtattctt aaaataagtt ggttccacac ataccaatac atgcatgtgc
     1561 tgattataag aattatcttt attatttatt gtcacttccg ttgcacgcat aaaaccaaca
     1621 agatttttat taattttttt atattgcatc attcggcgaa atccttgagc catatctgac
     1681 aaactcttat ttaattcttc gccatcataa acatttttaa ctgttaatgt gagaaacaac
     1741 caacgaactg ttggcttttg tttaataact tcagcaacaa ccttttgtga ctgaatgcca
     1801 tgtttcattg ctctcctcca gttgcacatt ggacaaagcc tggatttaca aaaccacact
     1861 cgatacaact ttctttcgcc tgtttcacga ttttgtttat actctaatat ttcagcacaa
     1921 tcttttactc tttcagcctt tttaaattca agaatatgca gaagttcaaa gtaatcaaca
     1981 ttagcgattt tcttttctct ccatggtctc acttttccac tttttgtctt gtccactaaa
     2041 acccttgatt tttcatctga ataaatgcta ctattaggac acataatatt aaaagaaacc
     2101 cccatctatt tagttatttg tttggtcact tataacttta acagatgggg tttttctgtg
     2161 caaccaattt taagggtttt caatacttta aaacacatac ataccaacac ttcaacgcac
     2221 ctttcagcaa ctaaaataaa aatgacgtta tttctatatg tatcaagata agaaagaaca
     2281 agttcaaaac catcaaaaaa agacaccttt tcaggtgctt tttttatttt ataaactcat
     2341 tccctgatct cgacttcgtt ctttttttac ctctcggtta tgagttagtt caaattcgtt
     2401 ctttttaggt tctaaatcgt gtttttcttg gaattgtgct gttttatcct ttaccttgtc
     2461 tacaaacccc ttaaaaacgt ttttaaaggc ttttaagccg tctgtacgtt ccttaagcat
     2521 acgatttagg tgacactata gaatagaagt atagtgccgt tcttttgagc ggcctattac
     2581 tcaccagtct tcacggggag ggctggatag taaaaggagg tgaaatgtac aatggtccaa
     2641 actagttcga agatctccgg agctcccggg atccggtacc ggccgcatgc taactagcta
     2701 accggttacc tatgccctat ctaccttgcg taggtagggt tctttttgtt gccggtccaa
     2761 gaattggagc taattaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga
     2821 acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggccgcgttg
     2881 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2941 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3001 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3061 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3121 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3181 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3241 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3301 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3361 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3421 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3481 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3541 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3601 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3661 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3721 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3781 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3841 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3901 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3961 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4021 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4081 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4141 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4201 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4261 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4321 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4381 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4441 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4501 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4561 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4621 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     4681 aataggcgta tcacgaggcc ctttcgtctt caagaattcc tgttataaaa aaaggatcaa
     4741 ttttgaactc tctcccaaag ttgatccctt aacgatttag aaatcccttt gagaatgttt
     4801 atatacattc aaggtaacca gccaactaat gacaatgatt cctgaaaaaa gtaataacaa
     4861 attactatac agataagttg actgatcaac ttccataggt aacaaccttt gatcaagtaa
     4921 gggtatggat aataaaccac ctacaattgc aatacctgtt ccctctgata aaaagctggt
     4981 aaagttaagc aaactcattc cagcaccagc ttcctgctgt ttcaagctac ttgaaacaat
     5041 tgttgatata actgttttgg tgaacgaaag cccacctaaa acaaatacga ttataattgt
     5101 catgaaccat gatgttgttt ctaaaagaaa ggaagcagtt aaaaagctaa cagaaagaaa
     5161 tgtaactccg atgtttaaca cgtataaagg acctcttcta tcaacaagta tcccaccaat
     5221 gtagccgaaa ataatgacac tcattgttcc ag
//
