LOCUS Exported 5234 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Shuttle vector E. coli/B.meg., encoding a promoter and a terminator for RNA polymerase of the phage K1E. ACCESSION . VERSION . KEYWORDS pPK1E-2+t 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5234 /organism="synthetic DNA construct" /mol_type="other DNA" 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(3522..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(4651..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 //