LOCUS       62056_2220        4952 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p641 LI-Esp3I, complete sequence.
ACCESSION   MK495749
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4952)
  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 4952)
  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 4952)
  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..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        195..225
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             279..935
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1280..1582
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     terminator      1659..1753
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1784..1812
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2001..2531
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            2745..2853
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2947..3535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3592..4422)
                     /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      4436..4787
                     /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(4860..4946)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgtac gagagacgaa agctgaacga
       61 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
      121 acataatact gtaaaacaca acatatccag tcactatggc ggcccctcga ggggccgcat
      181 taggcacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt
      241 aggatccgtc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
      301 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
      361 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
      421 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
      481 atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg
      541 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
      601 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
      661 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtgtcagcca
      721 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
      781 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
      841 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
      901 tacaacagta ctgcgatgag tggcagggcg gggcgtaatc tagaggatcc ggcttactaa
      961 aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg
     1021 atatgtatac ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac
     1081 agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg
     1141 gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa
     1201 aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac
     1261 gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta
     1321 tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat
     1381 ccccctggcc agtgcacgtc tgctgtcaga taaagtcccc cgtgaacttt acccggtggt
     1441 gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtttc
     1501 cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat
     1561 taacctgatg ttctggggaa tataaatgtc aggctccctt atacacagcc agtcgtctct
     1621 tctggtcgtg actgggaaaa ccctggcgac tagtcttgga ctcctgttga tagatccagt
     1681 aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt
     1741 tattggtgag aatccagggg tccccaataa ttacgattta aatttgacat aagcctgttc
     1801 ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     1861 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     1921 tgtacagcct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     1981 gatgttatgg agcagcaacg atgttacgca gcagcaacga tgttacgcag cagggcagtc
     2041 gccctaaaac aaagttaggt ggctcaagta tgggcatcat tcgcacatgt aggctcggcc
     2101 ctgaccaagt caaatccatg cgggctgctc ttgatctttt cggtcgtgag ttcggagatg
     2161 tagccaccta ctcccaacat cagccggact ccgattacct cgggaacttg ctccgtagta
     2221 agacattcat cgcgcttgct gccttcgacc aagaagcggt tgttggcgct ctcgcggctt
     2281 acgttctgcc caagtttgag cagccgcgta gtgagatcta tatctatgat ctcgcagtct
     2341 ccggagagca ccggaggcag ggcattgcca ccgcgctcat caatctcctc aagcatgagg
     2401 ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga ttacggtgac gatcccgcag
     2461 tggctctcta tacaaagttg ggcatacggg aagaagtgat gcactttgat atcgacccaa
     2521 gtaccgccac ctaacaattc gttcaagccg agatcggctt cccggccgcg gagttgttcg
     2581 gtaaattgga caacggtccg cgcgttgtcc ttttccgctg cataaccctg cttcggggtc
     2641 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
     2701 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
     2761 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
     2821 ctcaacggga atcctgctct gcgaggctgg ccgtaggccg gccgataatc tcatgaccaa
     2881 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2941 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3001 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3061 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3121 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3181 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3241 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3301 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3361 cgaagggaga aaggcggaca ggcatccggt aagcggcagg gtcggaacag gagagcgcac
     3421 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3481 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3541 cagcaacgcg gccgtgaaag gcaggccggt ccgtggtggc cacggcctct aggccagatc
     3601 cagcggcatc tgggttagtc gagcgcgggc cgcttcccat gtctcaccag ggcgagcctg
     3661 tttcgcgatc tcagcatctg aaatcttccc ggccttgcgc ttcgctgggg ccttacccac
     3721 cgccttggcg ggcttcttcg gtccaaaact gaacaacaga tgtgtgacct tgcgcccggt
     3781 ctttcgctgc gcccactcca cctgtagcgg gctgtgctcg ttgatctgcg tcacggctgg
     3841 atcaagcact cgcaacttga agtccttgat cgagggatac cggccttcca gttgaaacca
     3901 ctttcgcagc tggtcaattt ctatttcgcg ctggccgatg ctgtcccatt gcatgagcag
     3961 ctcgtaaagc ctgatcgcgt gggtgctgtc catcttggcc acgtcagcca aggcgtattt
     4021 ggtgaactgt ttggtgagtt ccgtcaggta cggcagcatg tctttggtga acctgagttc
     4081 tacacggccc tcaccctccc ggtagatgat tgtttgcacc cagccggtaa tcatcacact
     4141 cggtcttttc cccttgccat tgggctcttg ggttaaccgg acttcccgcc gtttcaggcg
     4201 cagggccgct tctttgagct ggttgtagga agattcgata gggacacccg ccatcgtcgc
     4261 tatgtcctcc gccgtcactg aatacatcac ttcatcggtg acaggctcgc tcctcttcac
     4321 ctggctaata caggccagaa cgatccgctg ttcctgaaca ctgaggcgat acgcggcctc
     4381 gaccagggca ttgcttttgt aaaccattgg gggtgaggcc acgttcgaca ttccttgtgt
     4441 ataaggggac actgtatctg cgtcccacaa tacaacaaat ccgtcccttt acaacaacaa
     4501 atccgtccct tcttaacaac aaatccgtcc cttaatggca acaaatccgt ccctttttaa
     4561 actctacagg ccacggatta cgtggcctgt agacgtccta aaaggtttaa aagggaaaag
     4621 gaagaaaagg gtggaaacgc aaaaaacgca ccactacgtg gccccgttgg ggccgcattt
     4681 gtgcccctga aggggcgggg gaggcgtctg ggcaatcccc gttttaccag tcccctatcg
     4741 ccgcctgaga gggcgcagga agcgagtaat cagggtatcg aggcggattc acccttggcg
     4801 tccaaccagc ggcaccagcg gcgcctgaga ggggcgcgcc cagctgtcta gggcggcgga
     4861 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     4921 ttcgactgag cctttcgttt tatttgatgc ct
//
