LOCUS       62056_2095        5487 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p421 LII F4-5, complete sequence.
ACCESSION   MK495757
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5487)
  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 5487)
  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 5487)
  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..5487
                     /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"
     CDS             complement(3199..4344)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(4737..5268)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5395..5481)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct agggaagaca atgtcgcggt
       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 tgtctgagtt 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 cggccgcgat gcaggtggct gctgaacccc cagccggaac
     3181 tgaccccaca aggccctagc gtttgcaatg caccaggtca tcattgaccc aggcgtgttc
     3241 caccaggccg ctgcctcgca actcttcgca ggcttcgccg acctgctcgc gccacttctt
     3301 cacgcgggtg gaatccgatc cgcacatgag gcggaaggtt tccagcttga gcgggtacgg
     3361 ctcccggtgc gagctgaaat agtcgaacat ccgtcgggcc gtcggcgaca gcttgcggta
     3421 cttctcccat atgaatttcg tgtagtggtc gccagcaaac agcacgacga tttcctcgtc
     3481 gatcaggacc tggcaacggg acgttttctt gccacggtcc aggacgcgga agcggtgcag
     3541 cagcgacacc gattccaggt gcccaacgcg gtcggacgtg aagcccatcg ccgtcgcctg
     3601 taggcgcgac aggcattcct cggccttcgt gtaataccgg ccattgatcg accagcccag
     3661 gtcctggcaa agctcgtaga acgtgaaggt gatcggctcg ccgatagggg tgcgcttcgc
     3721 gtactccaac acctgctgcc acaccagttc gtcatcgtcg gcccgcagct cgacgccggt
     3781 gtaggtgatc ttcacgtcct tgttgacgtg gaaaatgacc ttgttttgca gcgcctcgcg
     3841 cgggattttc ttgttgcgcg tggtgaacag ggcagagcgg gccgtgtcgt ttggcatcgc
     3901 tcgcatcgtg tccggccacg gcgcaatatc gaacaaggaa agctgcattt ccttgatctg
     3961 ctgcttcgtg tgtttcagca acgcggcctg cttggcttcg ctgacctgtt ttgccaggtc
     4021 ctcgccggcg gtttttcgct tcttggtcgt catagttcct cgcgtgtcga tggtcatcga
     4081 cttcgccaaa cctgccgcct cctgttcgag acgacgcgaa cgctccacgg cggccgatgg
     4141 cgcgggcagg gcagggggag ccagttgcac gctgtcgcgc tcgatcttgg ccgtagcttg
     4201 ctggactatc gagccgacgg actggaaggt ttcgcggggc gcacgcatga cggtgcggct
     4261 tgcgatggtt tcggcatcct cggcggaaaa ccccgcgtcg atcagttctt gcctgtatgc
     4321 cttccggtca aacgtccgat tcattcaccc tccttgcggg attgccccgg aattaattcc
     4381 ccggatcgat ccgtcgatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc
     4441 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc
     4501 ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa
     4561 gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     4621 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtggctgc catttttggg
     4681 gtgaggccgt tcgcggccga ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg
     4741 ggccgggagg gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag
     4801 ggcgcagccc tggttaaaaa caaggtttat aaatattggt ttaaaagcag gttaaaagac
     4861 aggttagcgg tggccgaaaa acgggcggaa acccttgcaa atgctggatt ttctgcctgt
     4921 ggacagcccc tcaaatgtca ataggtgcgc ccctcatctg tcagcactct gcccctcaag
     4981 tgtcaaggat cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag
     5041 ggcacttatc cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt
     5101 ttcgccgatt tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag cctgcccctc
     5161 atctgtcaac gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct
     5221 gtcagtgagg gccaagtttt ccgcgaggta tccacaacgc cggcggccct acatggctct
     5281 gctgtagtga gtgggttgcg ctccggcagc ggtcctgatc ccccgcagaa aaaaaggatc
     5341 tcaagaagat cctttgatct tttctacggc gcgcccagct gtctagggcg gcggatttgt
     5401 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
     5461 ctgagccttt cgttttattt gatgcct
//
