LOCUS Exported 6616 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Shuttle vector E. coli/B.meg.; encoding the signal peptide for protein secretion of the Bacillus megaterium protein YngK under control of the xylose inducible promoter . ACCESSION . VERSION . KEYWORDS pSPYngK-hp SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6616) AUTHORS Stammen S, Muller BK, Korneli C, Biedendieck R, Gamer M, Franco-Lara E, Jahn D TITLE High-yield intra- and extracellular protein production using Bacillus megaterium. JOURNAL Appl Environ Microbiol. 2010 Jun;76(12):4037-46. doi: 10.1128/AEM.00431-10. Epub 2010 Apr 30. PUBMED 20435764 REFERENCE 2 (bases 1 to 6616) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6616 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(109..969) /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 732..751 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(970..1074) /gene="bla" /label=AmpR promoter primer_bind 1142..1160 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" CDS complement(1238..2614) /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(2814..3818) /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 5808..5825 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" rep_origin complement(5966..6554) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6046..6065) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 61 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 121 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 181 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 241 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 301 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 361 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 421 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 481 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 541 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 601 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 661 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 721 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 781 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 841 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 901 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 961 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 1021 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 1081 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 1141 aaataggcgt atcacgaggc cctttcgtct tcaagaattc ctgttataaa aaaaggatca 1201 attttgaact ctctcccaaa gttgatccct taacgattta gaaatccctt tgagaatgtt 1261 tatatacatt caaggtaacc agccaactaa tgacaatgat tcctgaaaaa agtaataaca 1321 aattactata cagataagtt gactgatcaa cttccatagg taacaacctt tgatcaagta 1381 agggtatgga taataaacca cctacaattg caatacctgt tccctctgat aaaaagctgg 1441 taaagttaag caaactcatt ccagcaccag cttcctgctg tttcaagcta cttgaaacaa 1501 ttgttgatat aactgttttg gtgaacgaaa gcccacctaa aacaaatacg attataattg 1561 tcatgaacca tgatgttgtt tctaaaagaa aggaagcagt taaaaagcta acagaaagaa 1621 atgtaactcc gatgtttaac acgtataaag gacctcttct atcaacaagt atcccaccaa 1681 tgtagccgaa aataatgaca ctcattgttc cagggaaaat aattacactt ccgatttcgg 1741 cagtacttag ctggtgaaca tctttcatca tataaggaac catagagaca aaccctgcta 1801 ctgttccaaa tataattccc ccacaaagaa ctccaatcat aaaaggtata tttttcccta 1861 atccgggatc aacaaaagga tctgttactt tcctgatatg ttttacaaat atcaggaatg 1921 acagcacgct aacgataaga aaagaaatgc tatatgatgt tgtaaacaac ataaaaaata 1981 caatgcctac agacattagt ataattcctt tgatatcaaa atgacctttt atccttactt 2041 ctttctttaa taatttcata agaaacggaa cagtgataat tgttatcata ggaatgagta 2101 gaagatagga ccaatgaata taatgggcta tcattccacc aatcgctgga ccgactcctt 2161 ctcccatggc tactatcgat ccaataagac caaatgcttt acccctattt tcctttggaa 2221 tatagcgcgc aactacaacc attacgagtg ctggaaatgc agctgcacca gccccttgaa 2281 taaaacgagc cataataagt aaggaaaaga aagaatggcc aacaaaccca attaccgacc 2341 cgaaacaatt tattataatt ccaaatagga gtaacctttt gatgcctaat tgatcagata 2401 gctttccata tacagctgtt ccaatggaaa aggttaacat aaaggctgtg ttcacccagt 2461 ttgtactcgc aggtggttta ttaaaatcat ttgcaatatc aggtaatgag acgttcaaaa 2521 ccatttcatt taatacgcta aaaaaagata aaatgcaaag ccaaattaaa atttggttgt 2581 gtcgtaaatt cgattgtgaa taggatgtat tcacatttca ccctccaata atgagggcag 2641 acgtagttta tagggttaat gatacgcttc cctcttttaa ttgaaccctg ttacattcat 2701 tacacttcat aattaattcc tcctaaactt gattaaaaca ttttaccaca tataaactaa 2761 gttttaaatt cagtatttca tcacttatac aacaatatgg cccgtttgtt gaactactct 2821 ttaataaaat aatttttccg ttcccaattc cacattgcaa taatagaaaa tccatcttca 2881 tcggcttttt cgtcatcatc tgtatgaatc aaatcgcctt cttctgtgtc atcaaggttt 2941 aattttttat gtatttcttt taacaaacca ccataggaga ttaacctttt acggtgtaaa 3001 ccttcctcca aatcagacaa acgtttcaaa ttcttttctt catcatcggt cataaaatcc 3061 gtatccttta caggatattt tgcagtttcg tcaattgccg attgtatatc cgatttatat 3121 ttatttttcg gtcgaatcat ttgaactttt acatttggat catagtctaa tttcattgcc 3181 tttttccaaa attgaatcca ttgtttttga ttcacgtagt tttctgtatt cttaaaataa 3241 gttggttcca cacataccaa tacatgcatg tgctgattat aagaattatc tttattattt 3301 attgtcactt ccgttgcacg cataaaacca acaagatttt tattaatttt tttatattgc 3361 atcattcggc gaaatccttg agccatatct gacaaactct tatttaattc ttcgccatca 3421 taaacatttt taactgttaa tgtgagaaac aaccaacgaa ctgttggctt ttgtttaata 3481 acttcagcaa caaccttttg tgactgaatg ccatgtttca ttgctctcct ccagttgcac 3541 attggacaaa gcctggattt acaaaaccac actcgataca actttctttc gcctgtttca 3601 cgattttgtt tatactctaa tatttcagca caatctttta ctctttcagc ctttttaaat 3661 tcaagaatat gcagaagttc aaagtaatca acattagcga ttttcttttc tctccatggt 3721 ctcacttttc cactttttgt cttgtccact aaaacccttg atttttcatc tgaataaatg 3781 ctactattag gacacataat attaaaagaa acccccatct atttagttat ttgtttggtc 3841 acttataact ttaacagatg gggtttttct gtgcaaccaa ttttaagggt tttcaatact 3901 ttaaaacaca tacataccaa cacttcaacg cacctttcag caactaaaat aaaaatgacg 3961 ttatttctat atgtatcaag ataagaaaga acaagttcaa aaccatcaaa aaaagacacc 4021 ttttcaggtg ctttttttat tttataaact cattccctga tctcgacttc gttctttttt 4081 tacctctcgg ttatgagtta gttcaaattc gttcttttta ggttctaaat cgtgtttttc 4141 ttggaattgt gctgttttat cctttacctt gtctacaaac cccttaaaaa cgtttttaaa 4201 ggcttttaag ccgtctgtac gttccttaag atcaacagtg atatagtttg ctaacctttg 4261 cgttcactta actaacttat aggggtaaca cttaaaaaag aatcaataac gatagaaacc 4321 gctcctaaag caggtgcatt ttttcctaac gaagaaggca atagttcaca tttattgtct 4381 aaatgagaat ggactctaga agaaacttcg tttttaatcg tatttaaaac aatgggatga 4441 gattcaatta tatgatttct caagataaca gcttctatat caaatgtatt aaggatattg 4501 gttaatccaa ttccgatata aaagccaaag ttttgaagtg catttaacat ttctacatca 4561 tttttatttg cgcgttccac aatctctttt cgagaaatat tcttttcttc tttagagagc 4621 gaagccagta acgctttttc agaagcatat aattcccaac agcctcgatt tccacagctg 4681 catttgggtc cattaaaatc tatcgtcata tgacccattt ccccagaaaa accctgaaca 4741 cctttataca attcgttgtt aataacaagt ccagttccaa ttccgatatt aatactgatg 4801 taaacgatgt tttcatagtt ttttgtcata ccaaatactt tttcaccgta tgctcctgca 4861 ttagcttcat tttcaacaaa aaccggaaca ttaaactcac tctcaattaa aaactgcaaa 4921 tctttgatat tccaatttaa gttaggcatg aaaataattt gctgatgacg atctacaagg 4981 cctggaacac aaattcctat tccgactaga ccataagggg actcaggcat atgggttaca 5041 aaaccatgaa taagtgcaaa taaaatctct tttacttcac tagcggaaga actagacaag 5101 tcagaagtct tctcgagaat aatatttcct tctaagtcgg ttagaattcc gttaagatag 5161 tcgactccta tatcaatacc aatcgagtag cctgcattct tattaaaaac aagcattaca 5221 ggtcttctgc cgcctctaga ttgccctgcc ccaatttcaa aaataaaatc tttttcaagc 5281 agtgtattta cttgagagga gacagtagac ttgtttaatc ctgtaatctc agagagagtt 5341 gccctggaga caggggagtt cttcaaaatt tcatctaata ttaatttttg attcattttt 5401 tttactaaag cttgatctgc aatttgaata ataaccactc ctttgtttat ccaccgaact 5461 aagttggtgt tttttgaagc ttgaattaga tatttaaaag tatcatatct aatattataa 5521 ctaaattttc taaaaaaaac attgacataa acatttaatt aatatatgat gagataaagt 5581 tagtttattg gataaacaaa ctaactcaat taagatagtt gatggataaa cttgttcact 5641 taaatcaagg aggtgaatgt acaatgtata ttaaaaaatg tataggaagc atcttgtttt 5701 tactgctgtt ttgttcgagc gcgttacctg ctaaagcaga tactagttcg aagatctccg 5761 gagctcccgg gatccggtac cggccgcatg ccggcctgag aggatcgcat caccatcacc 5821 atcactaacc ggtccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca 5881 aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg 5941 catctcgggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 6001 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 6061 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 6121 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 6181 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 6241 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 6301 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 6361 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta 6421 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 6481 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 6541 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 6601 aaaactcacg ttaagg //