LOCUS Exported 4921 bp ds-DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pYUB28b, complete sequence. ACCESSION HQ247814 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4921) AUTHORS Bashiri G, Rehan AM, Greenwood DR, Dickson JM, Baker EN. TITLE Metabolic Engineering of Cofactor F(420) Production in Mycobacterium smegmatis JOURNAL PLoS ONE 5 (12), E15803 (2010) PUBMED 21209917 REFERENCE 2 (bases 1 to 4921) AUTHORS Bashiri G, Baker EN. TITLE Direct Submission JOURNAL Submitted (08-SEP-2010) School of Biological Sciences, The University of Auckland, 3a Symonds St., Auckland, Auckland 1010, New Zealand REFERENCE 3 (bases 1 to 4921) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..4921 /organism="Cloning vector pYUB28b" /mol_type="other DNA" /label=E. coli/Mycobacterium shuttle vector /note="E. coli/Mycobacterium shuttle vector" /db_xref="taxon:944548" misc_feature 1..4921 /note="contains hygromycin B resistance protein, OriE (E. coli origin of replication), OriM (Mycobacterial origin of replication), T7 promoter, T7 terminator, and multiple cloning site" promoter 61..79 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 80..104 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 119..141 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 160..177 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 187..204 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 208..240 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 289..306 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 373..420 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 457..885 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" rep_origin complement(3001..3589) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat 61 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 121 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc 181 agcggcctgg tgccgcgcgg cagccatatg gctagcatga ctggtggaca gcaaatgggt 241 cgggatccga attcgagctc cgtcgacaag cttgcggccg cactcgagca ccaccaccac 301 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc 361 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg 421 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa 481 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 541 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 601 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 661 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 721 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 781 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 841 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatatcta 901 gatttaaaga tctggtaccg cggccgcgcc acggatgcca ccacaagcac tacaacggag 961 ttcgccacgt acatcaccac aaccaccgat tctggcggtg agctccacga tattcagcgg 1021 aaatggcttg gtatcgacca agattcgtag aaccccgtct cgtctggctg gtattcaaaa 1081 cggacgcaac gaaacacgca acgagacagg catggcccaa accagaaaac tagcgtctac 1141 caggactttt acctgtccga cccgttgcaa cggaaccccc cacggaaccc ccgcgacacc 1201 cgctccccaa ttgcgttaga acagcggtgg attgtcggct tcgttgtggg ccttttgagc 1261 cgcttcctgt tctgccgcac gctctttcct cgcccgatag ccgagtcgct taacggtgtc 1321 cagatgcagc ccgaaatgtt tggccgtttg cggccaagag tggccctcgt cgtcgtgata 1381 ggcgcggatg cgttcgcggc gtgcagcctg ctcggcgaac cactcgctgc gttcctgcgc 1441 cacgagccgg acgacgtggc gttcggatag tccggtgatt cgagcgcctt cggcggcggt 1501 cacgcgccgc tttttgcgga cagtcggctg ccagttgtag ccgtcgctgt agccgtcgct 1561 gtagccgtcg ctcatagcaa tgcctccatg catggctgac gcggactttg cgcgccgcgc 1621 aactgtgctc gccgccgtgc gcgctgctgc gcccttccgc gagatggccg catggcgcgc 1681 actgagtgtg gcctcgtaga ccacgatccc gtccgcccaa atgcgcgact tggttgtgat 1741 ccaacgccaa atgctgttgg cgatggcgcg gacctcgctg tccggtagcg gtccgggaca 1801 cacgtcgttg cacggaaatt cggcgtttcg cgcgtggcac tcggcataga tcgcgcggcc 1861 gagtccgtcc acgttccggg tcggcaggta gatccgcatg agggcgggaa gataggccca 1921 caacctgacg gaatcgaaca gtgcgcaatt ccgccctagc ggcgtcggag ccgctttgta 1981 cgtggtctgc tgacgccagc gcggcggtgg catgttcgcg ccgagctcgg cctcgatgtg 2041 gctgagtgtg tagagatctg agtggagcca ttccgtttcc caggcgatgt ggccggggtt 2101 tttggtcatg aggcctgagt aactgcggtc gccatcgacg gcgcgccgaa ggccttcggc 2161 gcacgccgcc atgtatgcga gcggcttacg ccgcgcgtat tcggtgcgtg gaacaggggc 2221 gttgagtgcc cacactgcgt gtgcgtggcc gttggcgcga ttgcccacga tcgcgttggg 2281 cagcggatgg gacccccggg cgctgagcgc tcggagcgct gcgtctggat ggtctacgtc 2341 cacgaccagc aggtttgcca gcgctgttgg gttcgcctcg atgtaccggc ggcctagggc 2401 cgacgcgcgg ctttggcggt agatcccctc gagcagatcg tcgcttgcca gggaccagta 2461 cggcagccag agctgctcaa attcgtcggc gacgtggctc acgcttgata ataaaccacg 2521 attaatcacc ggtgtatggt ccgacacgag ctccaagtca gatatttcgc tgaggggcca 2581 ccccacaact gcacactccc ccgctctccc gtcgagccct gatgatgaaa caccagcgac 2641 agccgagcac ccccaaccac ctgtaccaac caggaggaac acatgcgtcg tttcgaggac 2701 gtttccgggc cgctaagagc cgctgtggcg gccgtacacg ccgccttaga cccgttagac 2761 cccctgccgc ctgaatgcgc gggtacgagc cacacagcac ccgaacttac ggagctggtg 2821 ggctcacgcc gtgcggccgc acgcgttgcg ctcggtcgtt cggctgcggc gagcggtatc 2881 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 2941 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 3001 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 3061 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 3121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 3181 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 3241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 3301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 3361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 3421 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 3481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 3541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 3601 gatcttttct acggggtctg acgctcagtg gaactagagc atgagattat caaaaaggat 3661 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 3721 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 3781 tctatttcgt tcatccatag ttgcctgact ccccgacgtg gccgaccagc ccgtcatcgt 3841 caacgcctga ccgcggtgcg gacaggccgt gtcgcgaccg gccgtgcgga attaagccgg 3901 cccgtaccct gtgaatagag gtccgctgtg acacaagaat ccctgttact tctcgaccgt 3961 attgattcgg atgattccta cgcgagcctg cggaacgacc aggaattctg ggagccgctg 4021 gcccgccgag ccctggagga gctcgggctg ccggtgccgc cggtgctgcg ggtgcccggc 4081 gagagcacca accccgtact ggtcggcgag cccgacccgg tgatcaagct gttcggcgag 4141 cactggtgcg gtccggagag cctcgcgtcg gagtcggagg cgtacgcggt cctggcggac 4201 gccccggtgc cggtgccccg cctcctcggc cgcggcgagc tgcggcccgg caccggagcc 4261 tggccgtggc cctacctggt gatgagccgg atgaccggca ccacctggcg gtccgcgatg 4321 gacggcacga ccgaccggaa cgcgctgctc gccctggccc gcgaactcgg ccgggtgctc 4381 ggccggctgc acagggtgcc gctgaccggg aacaccgtgc tcacccccca ttccgaggtc 4441 ttcccggaac tgctgcggga acgccgcgcg gcgaccgtcg aggaccaccg cgggtggggc 4501 tacctctcgc cccggctgct ggaccgcctg gaggactggc tgccggacgt ggacacgctg 4561 ctggccggcc gcgaaccccg gttcgtccac ggcgacctgc acgggaccaa catcttcgtg 4621 gacctggccg cgaccgaggt caccgggatc gtcgacttca ccgacgtcta tgcgggagac 4681 tcccgctaca gcctggtgca actgcatctc aacgccttcc ggggcgaccg cgagatcctg 4741 gccgcgctgc tcgacggggc gcagtggaag cggaccgagg acttcgcccg cgaactgctc 4801 gccttcacct tcctgcacga cttcgaggtg ttcgaggaga ccccgctgga tctctccggc 4861 ttcaccgatc cggaggaact ggcgcagttc ctctgggggc cgccggacac cgcccccggc 4921 g //