LOCUS       62056_13535        4591 bp DNA     circular SYN 01-SEP-2021
DEFINITION  Bifidobacterium-E. coli shuttle vector pKO403-lacZ-P15A DNA, 
            complete sequence.
ACCESSION   LC647193
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4591)
  AUTHORS   Hend A, Yuka O, Tomoya K, Kouta S, Izumi N, Tohru S.
  TITLE     Bifidobacterium-E. coli shuttle vector series, pKO403, for metabolic
            engineering
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4591)
  AUTHORS   Hend A, Yuka O, Tomoya K, Kouta S, Izumi N, Tohru S.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-AUG-2021) Contact:Altaib Hend Gifu University, Applied
            Biological Science; 1-1Yanagido, Gifu, Gifu 501-1193, Japan
REFERENCE   3  (bases 1 to 4591)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-AUG-2021) Contact:Altaib Hend Gifu University, Applied 
            Biological Science; 1-1Yanagido, Gifu, Gifu 501-1193, Japan"
FEATURES             Location/Qualifiers
     source          1..4591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..11
                     /label=sapI site
                     /note="sapI site"
     primer_bind     334..350
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(354..410)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(420..436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(444..460)
                     /label=lac operator
                     /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)."
     promoter        complement(468..498)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(513..534)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    638..648
                     /label=sapI site
                     /note="sapI site"
     misc_feature    662..2286
                     /label=Ori-Ts
                     /note="Ori-Ts"
     gene            1288..1978
                     /gene="repB"
                     /label=repB
     misc_feature    1288..1978
                     /gene="repB"
     misc_feature    2301..2534
                     /label=hup terminator
                     /note="hup terminator"
     rep_origin      complement(2833..3378)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     misc_feature    3448..4550
                     /label=spectinomycin resistance gene
                     /note="spectinomycin resistance gene"
ORIGIN
        1 acatgaagag cgtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
       61 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
      121 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      181 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      241 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      301 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagcttg
      361 catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaattc gtaatcatgg
      421 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      481 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
      541 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
      601 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttcgtaggg ccgccctcag
      661 ggccaccgtc gccaaggctc ttgaacacaa cgtcaggacc agcgatggca tgaactggaa
      721 cctccaggac ctcgtctacg aggcgctgag cgaggaatgg cgcaaaaggg acggcgagat
      781 cagcgaccca tgggccaacg acgaggcgga cggataccag ccgccctcat acgagccggt
      841 caaccccgaa cgcaggactc cccagacgcc ctccgatggc ctgatctgac gtccgaaaaa
      901 aggcgctgtg cgcccttttt aaatctttta taaatctttt tacattcttt tagcccctcc
      961 gcagccttac tctcccaacg ggtttcagcc gaaacctaca ccaaaagggg agcgaaccta
     1021 caccaaaagg ggagcgaacc tacaccaaaa ggggagcgaa cctacaccaa aaggggagct
     1081 atatacacct tttgttattt aaggtgcaag ttgtgctatg ctgaggccat gtccaatgag
     1141 atcgtgaagt tcagcaacca gttcaacaac gtcgcgctga agaagttcga cgccgtgcac
     1201 ctggacgtgc tcatggcgat cgcctcaggg gtgagggaga agggcacggc cacggtggag
     1261 ttctcgttcg aggagctgcg cggcctcatg cgattgagga agaacctgac caacaagcag
     1321 ctggccgaca agatcgtgca gacgaacgcg cgcctgctgg cgctgaacta catgttcgag
     1381 gattcgggca agatcatcca gttcgcgctg ttcacgaagt tcgtcaccga cccgcaggag
     1441 gcgactctcg cggttggggt caacgaggag ttcgcgttcc tgctcaacga cctgaccagc
     1501 cagttcacgc gcttcgagct ggccgagttc gccgacctca agatcaagta cgccaaggag
     1561 ttctaccgca gggccaagca gtaccgcagc tccggaatct ggaagatcgg ccgcgacgag
     1621 ttctgccgac tgcttggcgt tccaccgtcg gcaataaccc agacacgata tctgaatcag
     1681 aaggttcttc agccaattca ggaggagtgt gggcctctcc ttggcctgaa gatcgagcgc
     1741 cagtacgtga aacgcaggct gtcgggcttc gtgttcacat tcgcccgcga gacccctccg
     1801 gtgatcgacg ccaggcccgt ggaggcgagg aagacggacg gcgacggcaa gggccattgg
     1861 acgagcgttg ccggatacgg cgaggtgttc acgaccacgg cgttgttcga cgtgacggcc
     1921 gcccgggctc acttcgacgg caccgtggag gccggggaat gccgtttctg cgcgtttgac
     1981 gcgcgcaacc gcgaacatca tgcgcggaac gccggaaggc tgttctagcg gccgtgtccg
     2041 cgcctctggg gcggttgcgc ctgccatggg tcgttctgcc actgttcggc ctcacgctgg
     2101 tctgtgcgct gcctgatctc cctgagcagg tcggccttgg tcctgggggc gcttcgctcc
     2161 tcgaacgggc cgctctcccc caggtcctcg ggctcgctca ggtccaacgg ctcgtcaccg
     2221 gacaggctcg ggccggttct ctccctgtgc cgggttctcc gcctgtgcgc gttgttctgg
     2281 ccatgcgact ctagaggatc tccttctgct cgtagcgatt acttcgagca ttactgacga
     2341 caaagacccc gaccgagatg gtcggggtct ttttgttgtg gtgctgtgac gtgttgtcca
     2401 accgtattat tccggactag ttcagcgctt ccataagctg ctcgaatccc agggtgggct
     2461 tggcctcgaa gggatcgagg agtatggctt cccggcgaga tctcgcgccg aagtaatcgc
     2521 tacgagcaga aggagatccc cgggtactga gctcgaattc agcggagatg agcggagtgt
     2581 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     2641 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     2701 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     2761 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     2821 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     2881 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
     2941 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     3001 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     3061 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     3121 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     3181 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     3241 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     3301 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     3361 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctaggctcat
     3421 gagcccgaag tagtatatat gagtactgaa ttcccattaa ataataaaac aaaaaaatta
     3481 aaaaagtgtt tccaccattt tttcaatttt tttataattt ttttaatctg ttatttaaat
     3541 agtttatagt taaatttaca ttttcattag tccattcaat attctctcca agataactac
     3601 gaactgctaa caaaattctc tccctatgtt ctaatggaga agattcagcc actgcatttc
     3661 ccgcaatatc ttttggtatg attttacccg tgtccatagt taaaatcata cggcataaag
     3721 ttaatataga gttggtttca tcatcctgat aattatctat taattcctct gacgaatcca
     3781 taatggctcg tctcacatca gaaaatggaa tatcaggtag taattcctct aagtcataat
     3841 ttccgtatat tcttttattt tttcgttttg cttggtaaag cattatggtt aaatctgaat
     3901 ttaattcctt ctgaggaatg tatccttgtt cataaagctc ttgtaaccat tctccataaa
     3961 taaattcttg tttgggagga tgattccacg gtaccatttc ttgctgaata ataattgtta
     4021 attcaatata tcgtaagttg cttttatctc ctattttttt tgaaataggt ctaatttttt
     4081 gtataagtat ttctttactt tgatctgtca atggttcaga tacgacgact aaaaagtcaa
     4141 gatcactatt tggttttagt ccactctcaa ctcctgatcc aaacatgtaa gtaccaataa
     4201 ggttattttt taaatgtttc cgaagtattt ttttcacttt attaatttgt tcgtatgtat
     4261 tcaaatatat cctcctcact attttgatta gtacctattt tatatccata gttgttaatt
     4321 aaataaactt aatttagttt atttatagat ttcattggct tctaaatttt ttatctagat
     4381 aataattatt ttagttaatt ttattctaga ttatatatga tatgatcttt catttccata
     4441 aaactaaagt aagtgtaaac ctattcattg ttttaaaaat atctcttgcc agtcacgtta
     4501 cgttattagt tatagttatt ataacatgta ttcacgaacg aaaatcgacc ctgcagagta
     4561 ctgagctcaa gcttgcatgc aggcctgtcg a
//
