LOCUS       62056_2185        6882 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p451 LII F4-5, complete sequence.
ACCESSION   MK495775
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6882)
  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 6882)
  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 6882)
  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..6882
                     /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             3875..4663
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            4812..4920
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(4934..5782)
                     /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(5772..6608)
                     /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 tgagaagaca atgtcgcggt gagaccaaag 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 gacggtctca tgtctgagtt gtcttcagca gtcgtgactg ggaaaaccct ggcgactagt
     3541 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     3601 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc caataattac
     3661 gatttacgta tttaaatgaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     3721 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     3781 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     3841 gcagggcagt cgccctaaaa caaagttaaa catcatgagg gaagcggtga tcgccgaagt
     3901 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     3961 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     4021 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     4081 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     4141 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     4201 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     4261 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     4321 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     4381 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     4441 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     4501 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     4561 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     4621 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataagaca attgtccttt
     4681 tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt
     4741 ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa
     4801 cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc
     4861 actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg
     4921 taggccggcc tcagcctgcc gccttgggcc gggtgatgtc gtacttgccc gccgcgaact
     4981 cggttaccgt ccagcccagc gcgaccagct ccggcaacgc ctcgcgcacc cgctggcggc
     5041 gcttgcgcat ggtcgaacca ctggcctctg acggccagac atagccgcac aaggtatcta
     5101 tggaagcctt gccggttttg ccggggtcga tccagccaca cagccgctgg tgcagcaggc
     5161 gggcggtttc gctgtccagc gcccgcacct cgtccatgct gatgcgcaca tgctggccgc
     5221 cacccatgac ggcctgcgcg atcaaggggt tcagggccac gtacaggcgc ccgtccgcct
     5281 cgtcgctggc gtactccgac agcagccgaa acccctgccg cttgcggcca ttctgggcga
     5341 tgatggatac cttccaaagg cgctcgatgc agtcctgtat gtgcttgagc gccccaccac
     5401 tatcgacctc tgccccgatt tcctttgcca gcgcccgata gctacctttg accacatggc
     5461 attcagcggt gacggcctcc cacttgggtt ccaggaacag ccggagctgc cgtccgcctt
     5521 cggtcttggg ttccgggcca agcactaggc cattaggccc agccatggcc accagccctt
     5581 gcaggatgcg cagatcatca gcgcccagcg gctccgggcc gctgaactcg atccgcttgc
     5641 cgtcgccgta gtcatacgtc acgtccagct tgctgcgctt gcgctcgccc cgcttgaggg
     5701 cacggaacag gccgggggcc agacagtgcg ccgggtcgtg ccggacgtgg ctgaggctgt
     5761 gcttgttctt aggcttcacc acggggcacc cccttgctct tgcgctgcct ctccagcacg
     5821 gcgggcttga gcaccccgcc gtcatgccgc ctgaaccacc gatcagcgaa cggtgcgcca
     5881 tagttggcct tgctcacacc gaagcggacg aagaaccggc gctggtcgtc gtccacaccc
     5941 cattcctcgg cctcggcgct ggtcatgctc gacaggtagg actgccagcg gatgttatcg
     6001 accagtaccg agctgccccg gctggcctgc tgctggtcgc ctgcgcccat catggccgcg
     6061 cccttgctgg catggtgcag gaacacgata gagcacccgg tatcggcggc gatggcctcc
     6121 atgcgaccga tgacctgggc catggggccg ctggcgtttt cttcctcgat gtggaaccgg
     6181 cgcagcgtgt ccagcaccat caggcggcgg ccctcggcgg cgcgcttgag gccgtcgaac
     6241 cactccgggg ccatgatgtt gggcaggctg ccgatcagcg gctggatcag caggccgtca
     6301 gccacggctt gccgttcctc ggcgctgagg tgcgccccaa gggcgtgcag gcggtgatga
     6361 atggcggtgg gcggatcttc ggcgggcagg tagatcaccg ggccggtggg cagttcgccc
     6421 acctccagca gatccggccc gcctgcaatc tgtgcggcca gttgcagggc cagcatggat
     6481 ttaccggcac caccgggcga caccagcgcc ccgaccgtac cggccaccat gttgggcaaa
     6541 acgtagtcca gcggtggcgg cgctgctgcg aacgcctcca gaatattgat aggcttatgg
     6601 gtagccattg attgcctcct ttgcaggcag ttggtggtta ggcgctggcg gggtcactac
     6661 ccccgccctg cgccgctctg agttcttcca ggcactcgcg cagcgcctcg tattcgtcgt
     6721 cggtcagcca gaacttgcgc tgacgcatcc ctttggcctt catgcgctcg gcatatcgcg
     6781 cttggcgtac agcgtcaggg ctggccagca ggtcgccggt ctgcttgtcc ttttggtctt
     6841 tcatatcagt caccgagaaa cttgccgggg ccgaaaggct tg
//
