LOCUS       62056_1915        4728 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p231 LIII A-B, complete sequence.
ACCESSION   MK495765
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4728)
  AUTHORS   Chiasson D, Gimenez-Oya V, Bircheneder M, Bachmaier S, Studtrucker 
            T, Ryan J, Sollweck K, Leonhardt H, Boshart M, Dietrich P, Parniske 
            M.
  TITLE     A unified multi-kingdom Golden Gate cloning platform
  JOURNAL   Sci Rep 9 (1), 10131 (2019)
  PUBMED    31300661
REFERENCE   2  (bases 1 to 4728)
  AUTHORS   Chiasson D.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2019) Biology, LMU Munich, Grosshaderner Str. 2-4,
            Martinsried 82152, Germany
REFERENCE   3  (bases 1 to 4728)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep"; 
            date: "2019"; volume: "9"; issue: "1"; pages: "10131"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-FEB-2019) Biology, LMU Munich, Grosshaderner Str. 2-4, 
            Martinsried 82152, Germany"
FEATURES             Location/Qualifiers
     source          1..4728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(141..227)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        446..476
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             530..1186
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1531..1833
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     terminator      1993..2087
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2219..3031
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            3201..3309
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3323..3982)
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(join(3983..4728,1..26))
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 agcgaagcga gccagccggt ggccgctcgc ggccatcgtc cacatatcca cgggctggca
       61 agggagcgca gcgaccgcgc agggcgaagc ccggagagca agcccgtagg gggggcgcgc
      121 ccagctgtct agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      181 ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctttaatta
      241 aagcggataa caatttcaca caggaggccg cctaggggtc tcagcggtac gttgtcttca
      301 aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
      361 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg cggcccctcg
      421 aggggccgca ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg
      481 gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa
      541 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
      601 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
      661 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
      721 ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat
      781 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
      841 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
      901 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
      961 cgtgtcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     1021 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     1081 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     1141 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ctagaggatc
     1201 cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag
     1261 aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt
     1321 attacagtga cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata
     1381 tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct
     1441 ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct
     1501 cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga
     1561 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga
     1621 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtccc ccgtgaactt
     1681 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     1741 gtgccggttt ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     1801 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc
     1861 cagtctgcag gtcgacgaag actttctgtc tgtgagaccg agctcggtac ccggggatcc
     1921 tctagagtcg acctgcaggc atgcaagctt gcggccgcgt cgtgactggg aaaaccctgg
     1981 cgactagtct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt
     2041 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca ggggtcccca
     2101 ataattacga tttaaatttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2161 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2221 gagccatatt cagcgtgaaa cgagctgtag ccgtccgcgt ctgaacagca acatggatgc
     2281 ggatctgtat ggctataaat gggcgcgtga taacgtgggt cagagcggcg cgaccattta
     2341 tcgtctgtat ggcaaaccgg atgcgccgga actgtttctg aaacatggca aaggcagcgt
     2401 ggcgaacgat gtgaccgatg aaatggtgcg tctgaactgg ctgaccgaat ttatgccgct
     2461 gccgaccatt aaacatttta ttcgcacccc ggatgatgcg tggctgctga ccaccgcgat
     2521 tccgggcaaa accgcgtttc aggtgctgga agaatatccg gatagcggcg aaaacattgt
     2581 ggatgcgctg gccgtgtttc tgcgtcgtct gcatagcatt ccggtgtgca actgcccgtt
     2641 taacagcgat cgtgtgtttc gtctggccca ggcgcagagc cgtatgaaca acggcctggt
     2701 ggatgcgagc gattttgatg atgaacgtaa cggctggccg gtggaacagg tgtggaaaga
     2761 aatgcataaa ctgctgccgt ttagcccgga tagcgtggtg acccacggcg attttagcct
     2821 ggataacctg attttcgatg aaggcaaact gattggctgc attgatgtgg gccgtgtggg
     2881 cattgcggat cgttatcagg atctggccat tctgtggaac tgcctgggcg aatttagccc
     2941 gagcctgcaa aaacgtctgt ttcagaaata tggcattgat aatccggata tgaacaaact
     3001 gcaatttcat ctgatgctgg atgaattttt ctaataatta attggaccgc ggtccgcgcg
     3061 ttgtcctttt ccgctgcata accctgcttc ggggtcatta tagcgatttt ttcggtatat
     3121 ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg
     3181 tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc gagcgggtgt
     3241 tccttcttca ctgtccctta ttcgcacctg gcggtgctca acgggaatcc tgctctgcga
     3301 ggctggccgt aggccggccc taccggcgcg gcagcgttac ccgtgtcggc ggctccaacg
     3361 gctcgccatc gtccagaaaa cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc
     3421 gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg
     3481 gctggctggg cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg
     3541 tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa
     3601 ccaccacggc ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca
     3661 cgcggtcatt gaccacgtag gccgacacgg tgccggggcc gttgagcttc acgacggaga
     3721 tccagcgctc ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga
     3781 tgagcttgga aagtgttttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca
     3841 cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct
     3901 catgcgcttt gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga
     3961 tttctctgga ctgcgtggcc atgcttatct ccatgcggta ggggtgccgc acggttgcgg
     4021 caccatgcgc aatcagctgc aacttttcgg cagcgcgaca acaattatgc gttgcgtaaa
     4081 agtggcagtc aattacagat tttctttaac ctacgcaatg agctattgcg gggggtgccg
     4141 caatgagctg ttgcgtaccc ccctttttta agttgttgat ttttaagtct ttcgcatttc
     4201 gccctatatc tagttctttg gtgcccaaag aagggcaccc ctgcggggtt cccccacgcc
     4261 ttcggcgcgg ctccccctcc ggcaaaaagt ggcccctccg gggcttgttg atcgactgcg
     4321 cggccttcgg ccttgcccaa ggtggcgctg cccccttgga acccccgcac tcgccgccgt
     4381 gaggctcggg gggcaggcgg gcgggcttcg cccttcgact gcccccactc gcataggctt
     4441 gggtcgttcc aggcgcgtca aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc
     4501 gccttccact tggtgtccaa ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt
     4561 ccccagagaa aattaaaaaa attgatgggg caaggccgca ggccgcgcag ttggagccgg
     4621 tgggtatgtg gtcgaaggct gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc
     4681 aggcgcagcc tgtccatcag cttgtccagc agggttgtcc acgggccg
//
