LOCUS       62056_2140        5234 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p441 LII F1-2, complete sequence.
ACCESSION   MK495766
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5234)
  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 5234)
  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 5234)
  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..5234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        213..243
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             297..953
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1298..1600
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     terminator      1760..1854
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2090..2878
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            3027..3135
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      3229..3817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3874..4704)
                     /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      4718..5069
                     /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(5142..5228)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct agggaagaca atacggcggt
       61 gagaccaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt
      121 ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc actatggcgg
      181 cccctcgagg ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata
      241 atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa gctaaaatgg
      301 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
      361 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
      421 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
      481 ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc
      541 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
      601 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
      661 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
      721 tgtttttcgt gtcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
      781 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
      841 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
      901 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatcta
      961 gaggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg
     1021 gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa
     1081 gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat
     1141 gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc
     1201 gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg
     1261 aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta
     1321 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     1381 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtcccccg
     1441 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     1501 ggccagtgtg ccggtttccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     1561 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
     1621 acacagccag tctgcaggtc gacggtctca tgtcgggctt gtcttcgagc tcggtacccg
     1681 gggatcctct agagtcgacc tgcaggcatg caagcttgcg gccgcgtcgt gactgggaaa
     1741 accctggcga ctagtcttgg actcctgttg atagatccag taatgacctc agaactccat
     1801 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaggg
     1861 gtccccaata attacgattt acgtatttaa atgaaccttg accgaacgca gcggtggtaa
     1921 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc
     1981 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     2041 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgagggaagc
     2101 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     2161 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     2221 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     2281 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     2341 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     2401 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     2461 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     2521 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     2581 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     2641 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     2701 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     2761 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     2821 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     2881 agacaattgt ccttttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg
     2941 gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt
     3001 ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc
     3061 gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct
     3121 ctgcgaggct ggccgtaggc cggccgataa tctcatgacc aaaatccctt aacgtgagtt
     3181 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3241 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3301 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3361 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3421 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3481 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3541 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3601 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3661 caggcatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3721 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3781 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccgtgaa
     3841 aggcaggccg gtccgtggtg gccacggcct ctaggccaga tccagcggca tctgggttag
     3901 tcgagcgcgg gccgcttccc atgtctcacc agggcgagcc tgtttcgcga tctcagcatc
     3961 tgaaatcttc ccggccttgc gcttcgctgg ggccttaccc accgccttgg cgggcttctt
     4021 cggtccaaaa ctgaacaaca gatgtgtgac cttgcgcccg gtctttcgct gcgcccactc
     4081 cacctgtagc gggctgtgct cgttgatctg cgtcacggct ggatcaagca ctcgcaactt
     4141 gaagtccttg atcgagggat accggccttc cagttgaaac cactttcgca gctggtcaat
     4201 ttctatttcg cgctggccga tgctgtccca ttgcatgagc agctcgtaaa gcctgatcgc
     4261 gtgggtgctg tccatcttgg ccacgtcagc caaggcgtat ttggtgaact gtttggtgag
     4321 ttccgtcagg tacggcagca tgtctttggt gaacctgagt tctacacggc cctcaccctc
     4381 ccggtagatg attgtttgca cccagccggt aatcatcaca ctcggtcttt tccccttgcc
     4441 attgggctct tgggttaacc ggacttcccg ccgtttcagg cgcagggccg cttctttgag
     4501 ctggttgtag gaagattcga tagggacacc cgccatcgtc gctatgtcct ccgccgtcac
     4561 tgaatacatc acttcatcgg tgacaggctc gctcctcttc acctggctaa tacaggccag
     4621 aacgatccgc tgttcctgaa cactgaggcg atacgcggcc tcgaccaggg cattgctttt
     4681 gtaaaccatt gggggtgagg ccacgttcga cattccttgt gtataagggg acactgtatc
     4741 tgcgtcccac aatacaacaa atccgtccct ttacaacaac aaatccgtcc cttcttaaca
     4801 acaaatccgt cccttaatgg caacaaatcc gtcccttttt aaactctaca ggccacggat
     4861 tacgtggcct gtagacgtcc taaaaggttt aaaagggaaa aggaagaaaa gggtggaaac
     4921 gcaaaaaacg caccactacg tggccccgtt ggggccgcat ttgtgcccct gaaggggcgg
     4981 gggaggcgtc tgggcaatcc ccgttttacc agtcccctat cgccgcctga gagggcgcag
     5041 gaagcgagta atcagggtat cgaggcggat tcacccttgg cgtccaacca gcggcaccag
     5101 cggcgcctga gaggggcgcg cccagctgtc tagggcggcg gatttgtcct actcaggaga
     5161 gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt
     5221 tttatttgat gcct
//
