LOCUS       62056_17005        5535 bp DNA     circular SYN 22-JAN-2021
DEFINITION  Cloning vector pMJc01, complete sequence.
ACCESSION   MN201591
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5535)
  AUTHORS   Jutersek M, Dolinar M.
  TITLE     A new vector for cyanobacterial biotechnology assayed with cystatin,
            a distinct non-fluorescent reporter protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5535)
  AUTHORS   Jutersek M, Dolinar M.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUL-2019) Faculty of Chemistry and Chemical 
            Technology, Vecna pot 113, Ljubljana SI-1000, Slovenia
REFERENCE   3  (bases 1 to 5535)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-JUL-2019) Faculty of Chemistry and Chemical Technology, Vecna 
            pot 113, Ljubljana SI-1000, Slovenia"
FEATURES             Location/Qualifiers
     source          1..5535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(9..403)
                     /direction=LEFT
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     protein_bind    473..494
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        509..539
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    547..563
                     /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)."
     CDS             599..1567
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             1631..1843
                     /codon_start=1
                     /transl_table=11
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="OrfE"
                     /protein_id="QGN03459.1"
                     /translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
                     LNLDGCTLSLFREDKPFGPGKFLGD"
     CDS             1845..2051
                     /codon_start=1
                     /transl_table=11
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="OrfF"
                     /protein_id="QGN03460.1"
                     /translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
                     EALRECLEELRAAQGGGSDPASA"
     CDS             2081..2917
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             2907..3755
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
     CDS             3879..4691
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     terminator      complement(4852..4910)
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     misc_feature    complement(4911..4931)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(4935..4952)
                     /label=6xHis
                     /note="6xHis affinity tag"
     regulatory      complement(5310..5319)
                     /regulatory_class="ribosome_binding_site"
     protein_bind    complement(5363..5381)
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    complement(5388..5409)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     terminator      5412..5455
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 aactgtcacg aacccctgca ataactgtca cgaacccctg caataactgt cacgaacccc
       61 tgcaataact gtcacgcccc caaacctgca aacccagcag gggcgggggc tggcggggtg
      121 ttggaaaaat ccatccatga ttatctaaga ataatccact aggcgcggtt atcagcgccc
      181 ttgtggggcg ctgctgccct tgcccaatat gcccggccag aggccggata gctggtctat
      241 tcgctgcgct aggctacaca ccgccccacc gctgcgcggc agggggaaag gcgggcaaag
      301 cccgctaaac cccacaccaa accccgcaga aatacgctgg agcgctttta gccgctttag
      361 cggcctttcc ccctacccga agggtggggg cgcgtgtgca gccccgcagg gcctgtctcg
      421 gtcgatcatt cagcccggct catagatctg cgggcagtga gcgcaacgca attaatgtaa
      481 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtataatgt
      541 gtggaattgt gagcggataa caatttcaca caggatctag gatccggggg gtggcccgat
      601 gaagaacgac aggactttgc aggccatagg ccgacagctc aaggccatgg gctgtgagcg
      661 cttcgatatc ggcgtcaggg acgccaccac cggccagatg atgaaccggg aatggtcagc
      721 cgccgaagtg ctccagaaca cgccatggct caagcggatg aatgcccagg gcaatgacgt
      781 gtatatcagg cccgccgagc aggagcggca tggtctggtg ctggtggacg acctcagcga
      841 gtttgacctg gatgacatga aagccgaggg ccgggagcct gccctggtag tggaaaccag
      901 cccgaagaac tatcaggcat gggtcaaggt ggccgacgcc gcaggcggtg aacttcgggg
      961 gcagattgcc cggacgctgg ccagcgagta cgacgccgac ccggccagcg ccgacagccg
     1021 ccactatggc cgcttggcgg gcttcaccaa ccgcaaggac aagcacacca cccgcgccgg
     1081 ttatcagccg tgggtgctgc tgcgtgaatc caagggcaag accgccaccg ctggcccggc
     1141 gctggtgcag caggctggcc agcagatcga gcaggcccag cggcagcagg agaaggcccg
     1201 caggctggcc agcctcgaac tgcccgagcg gcagcttagc cgccaccggc gcacggcgct
     1261 ggacgagtac cgcagcgaga tggccgggct ggtcaagcgc ttcggtgatg acctcagcaa
     1321 gtgcgacttt atcgccgcgc agaagctggc cagccggggc cgcagtgccg aggaaatcgg
     1381 caaggccatg gccgaggcca gcccagcgct ggcagagcgc aagcccggcc acgaagcgga
     1441 ttacatcgag cgcaccgtca gcaaggtcat gggtctgccc agcgtccagc ttgcgcgggc
     1501 cgagctggca cgggcaccgg caccccgcca gcgaggcatg gacaggggcg ggccagattt
     1561 cagcatgtag tgcttgcgtt ggtactcacg cctgttatac tatgagtact cacgcacaga
     1621 agggggtttt atggaatacg aaaaaagcgc ttcagggtcg gtctacctga tcaaaagtga
     1681 caagggctat tggttgcccg gtggctttgg ttatacgtca aacaaggccg aggctggccg
     1741 cttttcagtc gctgatatgg ccagccttaa ccttgacggc tgcaccttgt ccttgttccg
     1801 cgaagacaag cctttcggcc ccggcaagtt tctcggtgac tgatatgaaa gaccaaaagg
     1861 acaagcagac cggcgacctg ctggccagcc ctgacgctgt acgccaagcg cgatatgccg
     1921 agcgcatgaa ggccaaaggg atgcgtcagc gcaagttctg gctgaccgac gacgaatacg
     1981 aggcgctgcg cgagtgcctg gaagaactca gagcggcgca gggcgggggt agtgaccccg
     2041 ccagcgccta accaccaact gcctgcaaag gaggcaatca atggctaccc ataagcctat
     2101 caatattctg gaggcgttcg cagcagcgcc gccaccgctg gactacgttt tgcccaacat
     2161 ggtggccggt acggtcgggg cgctggtgtc gcccggtggt gccggtaaat ccatgctggc
     2221 cctgcaactg gccgcacaga ttgcaggcgg gccggatctg ctggaggtgg gcgaactgcc
     2281 caccggcccg gtgatctacc tgcccgccga agacccgccc accgccattc atcaccgcct
     2341 gcacgccctt ggggcgcacc tcagcgccga ggaacggcaa gccgtggctg acggcctgct
     2401 gatccagccg ctgatcggca gcctgcccaa catcatggcc ccggagtggt tcgacggcct
     2461 caagcgcgcc gccgagggcc gccgcctgat ggtgctggac acgctgcgcc ggttccacat
     2521 cgaggaagaa aacgccagcg gccccatggc ccaggtcatc ggtcgcatgg aggccatcgc
     2581 cgccgatacc gggtgctcta tcgtgttcct gcaccatgcc agcaagggcg cggccatgat
     2641 gggcgcaggc gaccagcagc aggccagccg gggcagctcg gtactggtcg ataacatccg
     2701 ctggcagtcc tacctgtcga gcatgaccag cgccgaggcc gaggaatggg gtgtggacga
     2761 cgaccagcgc cggttcttcg tccgcttcgg tgtgagcaag gccaactatg gcgcaccgtt
     2821 cgctgatcgg tggttcaggc ggcatgacgg cggggtgctc aagcccgccg tgctggagag
     2881 gcagcgcaag agcaaggggg tgccccgtgg tgaagcctaa gaacaagcac agcctcagcc
     2941 acgtccggca cgacccggcg cactgtctgg cccccggcct gttccgtgcc ctcaagcggg
     3001 gcgagcgcaa gcgcagcaag ctggacgtga cgtatgacta cggcgacggc aagcggatcg
     3061 agttcagcgg cccggagccg ctgggcgctg atgatctgcg catcctgcaa gggctggtgg
     3121 ccatggctgg gcctaatggc ctagtgcttg gcccggaacc caagaccgaa ggcggacggc
     3181 agctccggct gttcctggaa cccaagtggg aggccgtcac cgctgatgcc atggtggtca
     3241 aaggtagcta tcgggcgctg gcaaaggaaa tcggggcaga ggtcgatagt ggtggggcgc
     3301 tcaagcacat acaggactgc atcgagcgcc tttggaaggt atccatcatc gcccagaatg
     3361 gccgcaagcg gcaggggttt cggctgctgt cggagtacgc cagcgacgag gcggacgggc
     3421 gcctgtacgt ggccctgaac cccttgatcg cgcaggccgt catgggtggc ggccagcatg
     3481 tgcgcatcag catggacgag gtgcgggcgc tggacagcga aaccgcccgc ctgctgcacc
     3541 agcggctgtg tggctggatc gaccccggca aaaccggcaa ggcttccata gataccttgt
     3601 gcggctatgt ctggccgtca gaggccagtg gttcgaccat gcgcaagcgc cgccagcggg
     3661 tgcgcgaggc gttgccggag ctggtcgcgc tgggctggac ggtaaccgag ttcgcggcgg
     3721 gcaagtacga catcacccgg cccaaggcgg caggctgacc cattcaacaa agccacgttg
     3781 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     3841 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     3901 cgtcttgctc caggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     3961 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     4021 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     4081 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     4141 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
     4201 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     4261 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     4321 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     4381 acgagcgtaa tggctggcct gttgaacaag tctggaagga aatgcataag cttttgccat
     4441 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     4501 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     4561 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     4621 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     4681 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     4741 cttgacggga ctcgagatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4801 gaccgagcgc agcgagtcag tgagcgagga agcctgcata acgcgaagta atcttttcgg
     4861 ttttaaagaa aaagggcagg gtggtgacac cttgcccgtt ttttgccgga ctgcagcggc
     4921 cgctactagt atcaatggtg gtgatgatgg tgctggcatt tgctttccag cagtttgatc
     4981 tggttcagcc acggaatgga gtaaacgacg aaggtgcagg tggtatattt cgccatttcc
     5041 ggttcgtcgt gaaattcgca gctttgcaga tcgcctgaac ttttcggaca ggtggtacgg
     5101 ccaatttcca cttgcaggat gtatttgatg ccgctaacca gctgacgttt tgcgctaata
     5161 acgcgaacga cgcgactgct atatttgtcg ttgctcgcgc ggttatattc tgccattgca
     5221 aattgcagag cgcgttgcag accttcgtcg ttttcgtcaa ccggaaccgg tgcgcccagc
     5281 agacgactac gatcttcact catctagtaa cctccactac tctagtagct cccattatat
     5341 ctatcactga tagggatgtc aatctctatc actgataggg agggcccctc tagaagcggc
     5401 cgcgaattcc agaaatcatc cttagcgaaa gctaaggatt ttttttatct gaaattctgc
     5461 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5521 ggtggcacca tggaa
//
