LOCUS       62056_1925        6823 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p251 LIII A-B, complete sequence.
ACCESSION   MK495777
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6823)
  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 6823)
  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 6823)
  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..6823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(240..1208)
                     /codon_start=1
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     promoter        complement(1232..1262)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     rep_origin      1318..1712
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     terminator      complement(1750..1836)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        2053..2083
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2137..2793
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3138..3440
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     terminator      3545..3639
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3771..4583
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            4753..4861
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(4875..5723)
                     /codon_start=1
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="VVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(5713..6549)
                     /codon_start=1
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
ORIGIN
        1 acttcgcgga acaaggacaa ggtgcagccg tcaaggttaa ggctggccat atcagcgact
       61 gaaaagcggc cagcctcggc cttgtttgac gtataaccaa agccaccggg caaccaatag
      121 cccttgtcac ttttgatcag gtagaccgac cctgaagcgc ttttttcgta ttccataaaa
      181 cccccttctg tgcgtgagta ctcatagtat aacaggcgtg agtaccaacg caagcactac
      241 atgctgaaat ctggcccgcc cctgtccatg cctcgctggc ggggtgccgg tgcccgtgcc
      301 agctcggccc gcgcaagctg gacgctgggc agacccatga ccttgctgac ggtgcgctcg
      361 atgtaatccg cttcgtggcc gggcttgcgc tctgccagcg ctgggctggc ctcggccatg
      421 gccttgccga tttcctcggc actgcggccc cggctggcca gcttctgcgc ggcgataaag
      481 tcgcacttgc tgaggtcatc accgaagcgc ttgaccagcc cggccatctc gctgcggtac
      541 tcgtccagcg ccgtgcgccg gtggcggcta agctgccgct cgggcagttc gaggctggcc
      601 agcctgcggg ccttctcctg ctgccgctgg gcctgctcga tctgctggcc agcctgctgc
      661 accagcgccg ggccagcggt ggcggtcttg cccttggatt cacgcagcag cacccacggc
      721 tgataaccgg cgcgggtggt gtgcttgtcc ttgcggttgg tgaagcccgc caagcggcca
      781 tagtggcggc tgtcggcgct ggccgggtcg gcgtcgtact cgctggccag cgtccgggca
      841 atctgccccc gaagttcacc gcctgcggcg tcggccacct tgacccatgc ctgatagttc
      901 ttcgggctgg tttccactac cagggcaggc tcccggccct cggctttcat gtcatccagg
      961 tcaaactcgc tgaggtcgtc caccagcacc agaccatgcc gctcctgctc ggcgggcctg
     1021 atatacacgt cattgccctg ggcattcatc cgcttgagcc atggcgtgtt ctggagcact
     1081 tcggcggctg accattcccg gttcatcatc tggccggtgg tggcgtccct gacgccgata
     1141 tcgaagcgct cacagcccat ggccttgagc tgtcggccta tggcctgcaa agtcctgtcg
     1201 ttcttcatcg ggccaccaag cgattcccac acattatacg agccggaagc ataaagtgta
     1261 aagcctagat ccgaaggatg agccgggctg aatgatcgac cgagacaggc cctgcggggc
     1321 tgcacacgcg cccccaccct tcgggtaggg ggaaaggccg ctaaagcggc taaaagcgct
     1381 ccagcgtatt tctgcggggt ttggtgtggg gtttagcggg ctttgcccgc ctttccccct
     1441 gccgcgcagc ggtggggcgg tgtgtagcct agcgcagcga atagaccagc tatccggcct
     1501 ctggccgggc atattgggca agggcagcag cgccccacaa gggcgctgat aaccgcgcct
     1561 agtggattat tcttagataa tcatggatgg atttttccaa caccccgcca gcccccgccc
     1621 ctgctgggtt tgcaggtttg ggggcgtgac agttattgca ggggttcgtg acagttattg
     1681 caggggggcg tgacagttat tgcaggggtt cgtgacagtt agggcgcgcc cagctgtcta
     1741 gggcggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     1801 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctttaattaa agcggataac
     1861 aatttcacac aggaggccgc tgaggtctca gcggtacgtt gtcttcaaag ctgaacgaga
     1921 aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac
     1981 ataatactgt aaaacacaac atatccagtc actatggcgg cccctcgagg ggccgcatta
     2041 ggcaccccag gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag
     2101 gatccgtcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     2161 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     2221 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     2281 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     2341 gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     2401 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     2461 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     2521 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt gtcagccaat
     2581 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     2641 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     2701 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     2761 caacagtact gcgatgagtg gcagggcggg gcgtaatcta gaggatccgg cttactaaaa
     2821 gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat
     2881 atgtataccc gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag
     2941 ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt
     3001 aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa
     3061 tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga
     3121 gaacaggggc tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc
     3181 gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc
     3241 ccctggccag tgcacgtctg ctgtcagata aagtcccccg tgaactttac ccggtggtgc
     3301 atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtttccg
     3361 ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta
     3421 acctgatgtt ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggtc
     3481 gacgaagact ttctgtctgt gagaccagca gtcgtgactg ggaaaaccct ggcgactagt
     3541 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     3601 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc caataattac
     3661 gatttaaatt tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     3721 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     3781 ttcagcgtga aacgagctgt agccgtccgc gtctgaacag caacatggat gcggatctgt
     3841 atggctataa atgggcgcgt gataacgtgg gtcagagcgg cgcgaccatt tatcgtctgt
     3901 atggcaaacc ggatgcgccg gaactgtttc tgaaacatgg caaaggcagc gtggcgaacg
     3961 atgtgaccga tgaaatggtg cgtctgaact ggctgaccga atttatgccg ctgccgacca
     4021 ttaaacattt tattcgcacc ccggatgatg cgtggctgct gaccaccgcg attccgggca
     4081 aaaccgcgtt tcaggtgctg gaagaatatc cggatagcgg cgaaaacatt gtggatgcgc
     4141 tggccgtgtt tctgcgtcgt ctgcatagca ttccggtgtg caactgcccg tttaacagcg
     4201 atcgtgtgtt tcgtctggcc caggcgcaga gccgtatgaa caacggcctg gtggatgcga
     4261 gcgattttga tgatgaacgt aacggctggc cggtggaaca ggtgtggaaa gaaatgcata
     4321 aactgctgcc gtttagcccg gatagcgtgg tgacccacgg cgattttagc ctggataacc
     4381 tgattttcga tgaaggcaaa ctgattggct gcattgatgt gggccgtgtg ggcattgcgg
     4441 atcgttatca ggatctggcc attctgtgga actgcctggg cgaatttagc ccgagcctgc
     4501 aaaaacgtct gtttcagaaa tatggcattg ataatccgga tatgaacaaa ctgcaatttc
     4561 atctgatgct ggatgaattt ttctaataat taattggacc gcggtccgcg cgttgtcctt
     4621 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     4681 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     4741 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     4801 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     4861 gtaggccggc ctcagcctgc cgccttgggc cgggtgatgt cgtacttgcc cgccgcgaac
     4921 tcggttaccg tccagcccag cgcgaccagc tccggcaacg cctcgcgcac ccgctggcgg
     4981 cgcttgcgca tggtcgaacc actggcctct gacggccaga catagccgca caaggtatct
     5041 atggaagcct tgccggtttt gccggggtcg atccagccac acagccgctg gtgcagcagg
     5101 cgggcggttt cgctgtccag cgcccgcacc tcgtccatgc tgatgcgcac atgctggccg
     5161 ccacccatga cggcctgcgc gatcaagggg ttcagggcca cgtacaggcg cccgtccgcc
     5221 tcgtcgctgg cgtactccga cagcagccga aacccctgcc gcttgcggcc attctgggcg
     5281 atgatggata ccttccaaag gcgctcgatg cagtcctgta tgtgcttgag cgccccacca
     5341 ctatcgacct ctgccccgat ttcctttgcc agcgcccgat agctaccttt gaccacatgg
     5401 cattcagcgg tgacggcctc ccacttgggt tccaggaaca gccggagctg ccgtccgcct
     5461 tcggtcttgg gttccgggcc aagcactagg ccattaggcc cagccatggc caccagccct
     5521 tgcaggatgc gcagatcatc agcgcccagc ggctccgggc cgctgaactc gatccgcttg
     5581 ccgtcgccgt agtcatacgt cacgtccagc ttgctgcgct tgcgctcgcc ccgcttgagg
     5641 gcacggaaca ggccgggggc cagacagtgc gccgggtcgt gccggacgtg gctgaggctg
     5701 tgcttgttct taggcttcac cacggggcac ccccttgctc ttgcgctgcc tctccagcac
     5761 ggcgggcttg agcaccccgc cgtcatgccg cctgaaccac cgatcagcga acggtgcgcc
     5821 atagttggcc ttgctcacac cgaagcggac gaagaaccgg cgctggtcgt cgtccacacc
     5881 ccattcctcg gcctcggcgc tggtcatgct cgacaggtag gactgccagc ggatgttatc
     5941 gaccagtacc gagctgcccc ggctggcctg ctgctggtcg cctgcgccca tcatggccgc
     6001 gcccttgctg gcatggtgca ggaacacgat agagcacccg gtatcggcgg cgatggcctc
     6061 catgcgaccg atgacctggg ccatggggcc gctggcgttt tcttcctcga tgtggaaccg
     6121 gcgcagcgtg tccagcacca tcaggcggcg gccctcggcg gcgcgcttga ggccgtcgaa
     6181 ccactccggg gccatgatgt tgggcaggct gccgatcagc ggctggatca gcaggccgtc
     6241 agccacggct tgccgttcct cggcgctgag gtgcgcccca agggcgtgca ggcggtgatg
     6301 aatggcggtg ggcggatctt cggcgggcag gtagatcacc gggccggtgg gcagttcgcc
     6361 cacctccagc agatccggcc cgcctgcaat ctgtgcggcc agttgcaggg ccagcatgga
     6421 tttaccggca ccaccgggcg acaccagcgc cccgaccgta ccggccacca tgttgggcaa
     6481 aacgtagtcc agcggtggcg gcgctgctgc gaacgcctcc agaatattga taggcttatg
     6541 ggtagccatt gattgcctcc tttgcaggca gttggtggtt aggcgctggc ggggtcacta
     6601 cccccgccct gcgccgctct gagttcttcc aggcactcgc gcagcgcctc gtattcgtcg
     6661 tcggtcagcc agaacttgcg ctgacgcatc cctttggcct tcatgcgctc ggcatatcgc
     6721 gcttggcgta cagcgtcagg gctggccagc aggtcgccgg tctgcttgtc cttttggtct
     6781 ttcatatcag tcaccgagaa acttgccggg gccgaaaggc ttg
//
