LOCUS       62056_2125        4787 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p431 LII F3-4, complete sequence.
ACCESSION   MK495762
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4787)
  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 4787)
  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 4787)
  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..4787
                     /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             2323..3111
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            3260..3368
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3382..4041)
                     /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(4042..4787,1..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 cctagggaag acaaattggc ggtgagacca
      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 gtcgacggtc tcatgtctgt cttgtcttcg 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 tttacgtatt taaatgaacc ttgaccgaac gcagcggtgg taacggcgca
     2161 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
     2221 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     2281 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatgaggga agcggtgatc
     2341 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
     2401 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
     2461 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     2521 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     2581 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     2641 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     2701 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     2761 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     2821 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     2881 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     2941 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     3001 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     3061 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataagacaat
     3121 tgtccttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
     3181 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
     3241 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
     3301 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
     3361 gctggccgta ggccggccct accggcgcgg cagcgttacc cgtgtcggcg gctccaacgg
     3421 ctcgccatcg tccagaaaac acggctcatc gggcatcggc aggcgctgct gcccgcgccg
     3481 ttcccattcc tccgtttcgg tcaaggctgg caggtctggt tccatgcccg gaatgccggg
     3541 ctggctgggc ggctcctcgc cggggccggt cggtagttgc tgctcgcccg gatacagggt
     3601 cgggatgcgg cgcaggtcgc catgccccaa cagcgattcg tcctggtcgt cgtgatcaac
     3661 caccacggcg gcactgaaca ccgacaggcg caactggtcg cggggctggc cccacgccac
     3721 gcggtcattg accacgtagg ccgacacggt gccggggccg ttgagcttca cgacggagat
     3781 ccagcgctcg gccaccaagt ccttgactgc gtattggacc gtccgcaaag aacgtccgat
     3841 gagcttggaa agtgttttct ggctgaccac cacggcgttc tggtggccca tctgcgccac
     3901 gaggtgatgc agcagcattg ccgccgtggg tttcctcgca ataagcccgg cccacgcctc
     3961 atgcgctttg cgttccgttt gcacccagtg accgggcttg ttcttggctt gaatgccgat
     4021 ttctctggac tgcgtggcca tgcttatctc catgcggtag gggtgccgca cggttgcggc
     4081 accatgcgca atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa
     4141 gtggcagtca attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc
     4201 aatgagctgt tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg
     4261 ccctatatct agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct
     4321 tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc
     4381 ggccttcggc cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg
     4441 aggctcgggg ggcaggcggg cgggcttcgc ccttcgactg cccccactcg cataggcttg
     4501 ggtcgttcca ggcgcgtcaa ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg
     4561 ccttccactt ggtgtccaac cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc
     4621 cccagagaaa attaaaaaaa ttgatggggc aaggccgcag gccgcgcagt tggagccggt
     4681 gggtatgtgg tcgaaggctg ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca
     4741 ggcgcagcct gtccatcagc ttgtccagca gggttgtcca cgggccg
//
