LOCUS       sequence-176092-        4901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mycobacteria shuttle vector - pYUB28b with a single change to the 
            MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4901)
  AUTHORS   Bashiri G, Rehan AM, Greenwood DR, Dickson JM, Baker EN
  TITLE     Metabolic engineering of cofactor F420 production in Mycobacterium 
            smegmatis.
  JOURNAL   PLoS One. 2010 Dec 29;5(12):e15803.
  PUBMED    21209917
REFERENCE   2  (bases 1 to 4901)
  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"; journalName: "PLoS One.";
            date: "2010-12-29"; volume: "5(12)"; pages: "e15803"
COMMENT     Sequence Label: pYUB28b-MCS-optimized
FEATURES             Location/Qualifiers
     source          1..4901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..80
                     /label=T7
                     /note="T7 promoter, forward primer"
     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)."
     CDS             153..170
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             180..197
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             201..233
                     /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             282..299
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(352..370)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      366..413
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      450..878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(537..556)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     747..768
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      complement(2981..3569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3061..3080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
       61 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
      121 tgtttaactt taagaagggc atgcgcagca gccatcatca tcatcatcac agcagcggcc
      181 tggtgccgcg cggcagccat atggctagca tgactggtgg acagcaaatg ggtcgggatc
      241 cgaattcgag ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact
      301 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
      361 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
      421 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
      481 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
      541 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
      601 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
      661 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
      721 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
      781 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
      841 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat ctagatttaa
      901 agatctggta ccgcggccgc gccacggatg ccaccacagg cactacaacg gagttcgcca
      961 cgtacatcac cacaaccacc gattctggcg gtgagctccc cgatattcag cggaaatggc
     1021 ttggtatcga ccaagattcg tagaaccccg tctcgtctgg ctggtattca aaacgggcgc
     1081 aacgaaacac gcaacgagac aggcatggcc caaaccagaa aactagcgtc taccaggact
     1141 tttacctgtc cgacccgttg caacggaacc ccccacggaa cccccgcgac acccgctccc
     1201 caattgcgtt agaacagcgg tggattgtcg gcttcgttgt gggccttttg agccgcttcc
     1261 tgttctgccg cacgctcttt cctcgcccga tagccgagtc gcttaacggt gtccagatgc
     1321 agcccgaaat gtttggccgt ttgcggccaa gagtggccct cgtcgtcgtg ataggcgcgg
     1381 atgcgttcgc ggcgtgcagc ctgctcggcg agccactcgc tgcgttcctg cgccacgagc
     1441 cggacgacgt ggcgttcgga tagtccggtg attcgagcgc cttcggcggc ggtcacgcgc
     1501 cgctttttgc ggacagtcgg ctgccggttg tagccgtcgc tgtagccgtc gctcatagca
     1561 atgcctccat gcatggctga cgcggacttt gcgcgccgcg caactgtgct cgccgccgtg
     1621 cgcgctgctg cgcccttccg cgagatggcc gactggcgcg cactgagtgt ggcctcgtag
     1681 accacgatcc cgtccgccca aatgcgcgac ttggttgtga tccaacgcca aatgctgttg
     1741 gcgatggcgc ggacctcgct gtccggtagc ggtccgggac acacgtcgtt gcacggaaat
     1801 tcggcgtttc gcgcgtggca ctcggcatag atcgcgcggc cgagtccgtc cacgttccgg
     1861 gtcggcaggt agatccgcat gagggcggga cgataggccc acaacctgac ggaatcgaac
     1921 agtgcgcaat tccgccctag cggcgtcgga gccgctttgt acgtggtctg ctgacgccag
     1981 cgcggcggtg gcatgttcgc gccgagctcg gcctcgatgt ggctgagtgt gtagagatct
     2041 gagtggagcc attccgtttc ccaggcgatg tggccggggt ttttggtcat gaggcctgag
     2101 taactgcggt cgccatcgac ggcgcgccga aggccttcgg cgcacgccgc catgtatgcg
     2161 agcggcttac gccgcgcgta ttcggtgcgt ggaacagggg cgttgagtgc ccacactgcg
     2221 tgtgcgtggc cgttggcgcg attgcccacg atcgcgttgg gcagcggatg ggacccccgg
     2281 gcgctgagcg ctcggagcgc tgcgtctgga tggtctacgt ccacgaccag caggtttgcc
     2341 agcgctgttg ggttcgcctc gatgtaccgg cggcctaggg ccgacgcgcg gctttggcgg
     2401 tagatcccct cgagcagatc gtcgcttgcc agcggccagt acggcagcca gagctgctca
     2461 aattcgtcgg cgacgtggct cacgcttggt agtagaccac gattaatcac cggtgtatgg
     2521 tccgacacga gctccaagtc agatatttcg ctgaggggcc accccacaac tgcacactcc
     2581 cccgctctcc cgtcgagccc tggtggtgga acaccagcga cagccgagca cccccaacca
     2641 cctgtaccaa ccaggaggaa cacatgcgtc gtttcgagga cgtttccggg ccgctgagag
     2701 ccgctgtggc ggccgtacac gccgccttag acccgttaga ccccctgccg cctgaatgcg
     2761 cgggtacgag ccacacagcg cccgaactta cggagctggt gggctcacgc gtgcggccgc
     2821 acgcgttgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2881 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2941 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3001 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3061 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3121 cgcttaccgg atacctgtcc gcctttctcc cctcgggaag cgtggcgctt tctcatagct
     3181 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3241 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3301 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3361 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3421 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3481 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3541 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3601 acgctcatgg aactagagca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3661 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3721 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3781 tgcctgactc cccgacgtgg ccgaccagcc cgtcatcgtc aacgcctgat ccgcggtgcg
     3841 gacaggccgt gtcgcgaccg gccgtgcgga attaagccgg cccgtaccct gtgaatagag
     3901 gtccgctgtg acacaagaat ccctgttact tctcgaccgt attgattcgg atgattccta
     3961 cgcgagcctg cggaacgacc aggaattctg ggagccgctg gcccgccgag ccctggagga
     4021 gctcgggctg ccggtgccgc cggtgctgcg ggtgcccggc gagagcacca accccgtact
     4081 ggtcggcgag cccggcccgg tgatcaagct gttcggcgag cactggtgcg gtccggagag
     4141 cctcgcgtcg gagtcggagg cgtacgcggt cctggcggac gccccggtgc cggtgccccg
     4201 cctcctcggc cgcggcgagc tgcggcccgg caccggagcc tggccgtggc cctacctggt
     4261 gatgagccgg atgaccggca ccacctggcg gtccgcgatg gacggcacga ccgaccggaa
     4321 cgcgctgctc gccctggccc gcgaactcgg ccgggtgctc ggccggctgc acagggtgcc
     4381 gctgaccggg aacaccgtgc tcacccccca ttccgaggtc ttcccggaac tgctgcggga
     4441 acgccgcgcg gcgaccgtcg aggaccaccg cgggtggggc tacctctcgc cccggctgct
     4501 ggaccgcctg gaggactggc tgccggacgt ggacacgctg ctggccggcc gcgaaccccg
     4561 gttcgtccac ggcgacctgc acgggaccaa catcttcgtg gacctggccg cgaccgaggt
     4621 caccgggatc gtcgacttca ccgacgtcta tgcgggagac tcccgctaca gcctggtgca
     4681 actgcatctc aacgccttcc ggggcgaccg cgagatcctg gccgcgctgc tcgacggggc
     4741 gcagtggaag cggaccgagg acttcgcccg cgaactgctc gccttcacct tcctgcacga
     4801 cttcgaggtg ttcgaggaga ccccgctgga tctctccggc ttcaccgatc cggaggaact
     4861 ggcgcagttc ctctgggggc cgccggacac cgcccccggc g
//
