LOCUS       62056_1910        5428 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p221 LIII A-B, complete sequence.
ACCESSION   MK495759
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5428)
  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 5428)
  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 5428)
  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..5428
                     /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             1986..2798
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2968..3076
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3140..4285)
                     /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(4678..5209)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5336..5422)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggggtctca gcggtacgtt
       61 gtcttcaaag 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 gacgaagact ttctgtctgt gagaccgagc 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 aaatttgtgt ctcaaaatct ctgatgttac attgcacaag
     1921 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1981 gtgttatgag ccatattcag cgtgaaacga gctgtagccg tccgcgtctg aacagcaaca
     2041 tggatgcgga tctgtatggc tataaatggg cgcgtgataa cgtgggtcag agcggcgcga
     2101 ccatttatcg tctgtatggc aaaccggatg cgccggaact gtttctgaaa catggcaaag
     2161 gcagcgtggc gaacgatgtg accgatgaaa tggtgcgtct gaactggctg accgaattta
     2221 tgccgctgcc gaccattaaa cattttattc gcaccccgga tgatgcgtgg ctgctgacca
     2281 ccgcgattcc gggcaaaacc gcgtttcagg tgctggaaga atatccggat agcggcgaaa
     2341 acattgtgga tgcgctggcc gtgtttctgc gtcgtctgca tagcattccg gtgtgcaact
     2401 gcccgtttaa cagcgatcgt gtgtttcgtc tggcccaggc gcagagccgt atgaacaacg
     2461 gcctggtgga tgcgagcgat tttgatgatg aacgtaacgg ctggccggtg gaacaggtgt
     2521 ggaaagaaat gcataaactg ctgccgttta gcccggatag cgtggtgacc cacggcgatt
     2581 ttagcctgga taacctgatt ttcgatgaag gcaaactgat tggctgcatt gatgtgggcc
     2641 gtgtgggcat tgcggatcgt tatcaggatc tggccattct gtggaactgc ctgggcgaat
     2701 ttagcccgag cctgcaaaaa cgtctgtttc agaaatatgg cattgataat ccggatatga
     2761 acaaactgca atttcatctg atgctggatg aatttttcta ataattaatt ggaccgcggt
     2821 ccgcgcgttg tccttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc
     2881 ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt
     2941 tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag
     3001 cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc
     3061 tctgcgaggc tggccgtagg ccggccgcga tgcaggtggc tgctgaaccc ccagccggaa
     3121 ctgaccccac aaggccctag cgtttgcaat gcaccaggtc atcattgacc caggcgtgtt
     3181 ccaccaggcc gctgcctcgc aactcttcgc aggcttcgcc gacctgctcg cgccacttct
     3241 tcacgcgggt ggaatccgat ccgcacatga ggcggaaggt ttccagcttg agcgggtacg
     3301 gctcccggtg cgagctgaaa tagtcgaaca tccgtcgggc cgtcggcgac agcttgcggt
     3361 acttctccca tatgaatttc gtgtagtggt cgccagcaaa cagcacgacg atttcctcgt
     3421 cgatcaggac ctggcaacgg gacgttttct tgccacggtc caggacgcgg aagcggtgca
     3481 gcagcgacac cgattccagg tgcccaacgc ggtcggacgt gaagcccatc gccgtcgcct
     3541 gtaggcgcga caggcattcc tcggccttcg tgtaataccg gccattgatc gaccagccca
     3601 ggtcctggca aagctcgtag aacgtgaagg tgatcggctc gccgataggg gtgcgcttcg
     3661 cgtactccaa cacctgctgc cacaccagtt cgtcatcgtc ggcccgcagc tcgacgccgg
     3721 tgtaggtgat cttcacgtcc ttgttgacgt ggaaaatgac cttgttttgc agcgcctcgc
     3781 gcgggatttt cttgttgcgc gtggtgaaca gggcagagcg ggccgtgtcg tttggcatcg
     3841 ctcgcatcgt gtccggccac ggcgcaatat cgaacaagga aagctgcatt tccttgatct
     3901 gctgcttcgt gtgtttcagc aacgcggcct gcttggcttc gctgacctgt tttgccaggt
     3961 cctcgccggc ggtttttcgc ttcttggtcg tcatagttcc tcgcgtgtcg atggtcatcg
     4021 acttcgccaa acctgccgcc tcctgttcga gacgacgcga acgctccacg gcggccgatg
     4081 gcgcgggcag ggcaggggga gccagttgca cgctgtcgcg ctcgatcttg gccgtagctt
     4141 gctggactat cgagccgacg gactggaagg tttcgcgggg cgcacgcatg acggtgcggc
     4201 ttgcgatggt ttcggcatcc tcggcggaaa accccgcgtc gatcagttct tgcctgtatg
     4261 ccttccggtc aaacgtccga ttcattcacc ctccttgcgg gattgccccg gaattaattc
     4321 cccggatcga tccgtcgatc ttgatcccct gcgccatcag atccttggcg gcaagaaagc
     4381 catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc
     4441 cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa gcccactgca
     4501 agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
     4561 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtggctg ccatttttgg
     4621 ggtgaggccg ttcgcggccg aggggcgcag cccctggggg gatgggaggc ccgcgttagc
     4681 gggccgggag ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca
     4741 gggcgcagcc ctggttaaaa acaaggttta taaatattgg tttaaaagca ggttaaaaga
     4801 caggttagcg gtggccgaaa aacgggcgga aacccttgca aatgctggat tttctgcctg
     4861 tggacagccc ctcaaatgtc aataggtgcg cccctcatct gtcagcactc tgcccctcaa
     4921 gtgtcaagga tcgcgcccct catctgtcag tagtcgcgcc cctcaagtgt caataccgca
     4981 gggcacttat ccccaggctt gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt
     5041 tttcgccgat ttgcgaggct ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct
     5101 catctgtcaa cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc
     5161 tgtcagtgag ggccaagttt tccgcgaggt atccacaacg ccggcggccc tacatggctc
     5221 tgctgtagtg agtgggttgc gctccggcag cggtcctgat cccccgcaga aaaaaaggat
     5281 ctcaagaaga tcctttgatc ttttctacgg cgcgcccagc tgtctagggc ggcggatttg
     5341 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
     5401 actgagcctt tcgttttatt tgatgcct
//
