LOCUS       62056_2215        4914 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p641 LI-BpiI, complete sequence.
ACCESSION   MK495748
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4914)
  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 4914)
  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 4914)
  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..4914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        85..115
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             169..825
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1170..1472
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     terminator      1621..1715
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1746..1774
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1963..2493
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            2707..2815
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2909..3497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3554..4384)
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      4398..4749
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     terminator      complement(4822..4908)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggtacgaag tcttcctcga
       61 ggggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg
      121 attttgagtt aggatccgtc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
      181 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
      241 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
      301 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
      361 cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata
      421 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
      481 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
      541 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
      601 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
      661 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
      721 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
      781 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatc tagaggatcc
      841 ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga
      901 atatatactg atatgtatac ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta
      961 ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat
     1021 ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg
     1081 gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc
     1141 ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc tataaaagag
     1201 agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac
     1261 ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtcccc cgtgaacttt
     1321 acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg
     1381 tgccggtttc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca
     1441 aaaacgccat taacctgatg ttctggggaa tataaatgtc aggctccctt atacacagcc
     1501 agtctgcagg tcgacgaaga ctttctggag ctcggtaccc ggggatcctc tagagtcgac
     1561 ctgcaggcat gcaagcttgc ggccgcgtcg tgactgggaa aaccctggcg actagtcttg
     1621 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
     1681 tcggttgccg ccgggcgttt tttattggtg agaatccagg ggtccccaat aattacgatt
     1741 taaatttgac ataagcctgt tcggttcgta aactgtaatg caagtagcgt atgcgctcac
     1801 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
     1861 tggcttgtta tgactgtttt tttgtacagc ctatgcctcg ggcatccaag cagcaagcgc
     1921 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagcaac
     1981 gatgttacgc agcagggcag tcgccctaaa acaaagttag gtggctcaag tatgggcatc
     2041 attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc tcttgatctt
     2101 ttcggtcgtg agttcggaga cgtagccacc tactcccaac atcagccgga ctccgattac
     2161 ctcgggaact tgctccgtag taagacattc atcgcgcttg ctgccttcga ccaagaagcg
     2221 gttgttggcg ctctcgcggc ttacgttctg cccaagtttg agcagccgcg tagtgagatc
     2281 tatatctatg atctcgcagt ctccggagag caccggaggc agggcattgc caccgcgctc
     2341 atcaatctcc tcaagcatga ggccaacgcg cttggtgctt atgtgatcta cgtgcaagca
     2401 gattacggtg acgatcccgc agtggctctc tatacaaagt tgggcatacg ggaagaagtg
     2461 atgcactttg atatcgaccc aagtaccgcc acctaacaat tcgttcaagc cgagatcggc
     2521 ttcccggccg cggagttgtt cggtaaattg gacaacggtc cgcgcgttgt ccttttccgc
     2581 tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc
     2641 acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg
     2701 tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt
     2761 cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccgtaggc
     2821 cggccgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2881 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2941 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3001 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3061 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3121 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3181 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3241 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3301 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggcatccg gtaagcggca
     3361 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3421 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3481 ggcggagcct atggaaaaac gccagcaacg cggccgtgaa aggcaggccg gtccgtggtg
     3541 gccacggcct ctaggccaga tccagcggca tctgggttag tcgagcgcgg gccgcttccc
     3601 atgtctcacc agggcgagcc tgtttcgcga tctcagcatc tgaaatcttc ccggccttgc
     3661 gcttcgctgg ggccttaccc accgccttgg cgggcttctt cggtccaaaa ctgaacaaca
     3721 gatgtgtgac cttgcgcccg gtctttcgct gcgcccactc cacctgtagc gggctgtgct
     3781 cgttgatctg cgtcacggct ggatcaagca ctcgcaactt gaagtccttg atcgagggat
     3841 accggccttc cagttgaaac cactttcgca gctggtcaat ttctatttcg cgctggccga
     3901 tgctgtccca ttgcatgagc agctcgtaaa gcctgatcgc gtgggtgctg tccatcttgg
     3961 ccacgtcagc caaggcgtat ttggtgaact gtttggtgag ttccgtcagg tacggcagca
     4021 tgtctttggt gaacctgagt tctacacggc cctcaccctc ccggtagatg attgtttgca
     4081 cccagccggt aatcatcaca ctcggtcttt tccccttgcc attgggctct tgggttaacc
     4141 ggacttcccg ccgtttcagg cgcagggccg cttctttgag ctggttgtag gaagattcga
     4201 tagggacacc cgccatcgtc gctatgtcct ccgccgtcac tgaatacatc acttcatcgg
     4261 tgacaggctc gctcctcttc acctggctaa tacaggccag aacgatccgc tgttcctgaa
     4321 cactgaggcg atacgcggcc tcgaccaggg cattgctttt gtaaaccatt gggggtgagg
     4381 ccacgttcga cattccttgt gtataagggg acactgtatc tgcgtcccac aatacaacaa
     4441 atccgtccct ttacaacaac aaatccgtcc cttcttaaca acaaatccgt cccttaatgg
     4501 caacaaatcc gtcccttttt aaactctaca ggccacggat tacgtggcct gtagacgtcc
     4561 taaaaggttt aaaagggaaa aggaagaaaa gggtggaaac gcaaaaaacg caccactacg
     4621 tggccccgtt ggggccgcat ttgtgcccct gaaggggcgg gggaggcgtc tgggcaatcc
     4681 ccgttttacc agtcccctat cgccgcctga gagggcgcag gaagcgagta atcagggtat
     4741 cgaggcggat tcacccttgg cgtccaacca gcggcaccag cggcgcctga gaggggcgcg
     4801 cccagctgtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     4861 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcct
//
