LOCUS       62056_1920        5175 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p241 LII A-B, complete sequence.
ACCESSION   MK495771
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5175)
  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 5175)
  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 5175)
  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..5175
                     /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"
     rep_origin      3170..3758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3815..4645)
                     /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      4659..5010
                     /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(5083..5169)
                     /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 ccggccgata atctcatgac caaaatccct taacgtgagt
     3121 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3181 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3241 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3301 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3361 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3421 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3481 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3541 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3601 acaggcatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3661 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3721 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggccgtga
     3781 aaggcaggcc ggtccgtggt ggccacggcc tctaggccag atccagcggc atctgggtta
     3841 gtcgagcgcg ggccgcttcc catgtctcac cagggcgagc ctgtttcgcg atctcagcat
     3901 ctgaaatctt cccggccttg cgcttcgctg gggccttacc caccgccttg gcgggcttct
     3961 tcggtccaaa actgaacaac agatgtgtga ccttgcgccc ggtctttcgc tgcgcccact
     4021 ccacctgtag cgggctgtgc tcgttgatct gcgtcacggc tggatcaagc actcgcaact
     4081 tgaagtcctt gatcgaggga taccggcctt ccagttgaaa ccactttcgc agctggtcaa
     4141 tttctatttc gcgctggccg atgctgtccc attgcatgag cagctcgtaa agcctgatcg
     4201 cgtgggtgct gtccatcttg gccacgtcag ccaaggcgta tttggtgaac tgtttggtga
     4261 gttccgtcag gtacggcagc atgtctttgg tgaacctgag ttctacacgg ccctcaccct
     4321 cccggtagat gattgtttgc acccagccgg taatcatcac actcggtctt ttccccttgc
     4381 cattgggctc ttgggttaac cggacttccc gccgtttcag gcgcagggcc gcttctttga
     4441 gctggttgta ggaagattcg atagggacac ccgccatcgt cgctatgtcc tccgccgtca
     4501 ctgaatacat cacttcatcg gtgacaggct cgctcctctt cacctggcta atacaggcca
     4561 gaacgatccg ctgttcctga acactgaggc gatacgcggc ctcgaccagg gcattgcttt
     4621 tgtaaaccat tgggggtgag gccacgttcg acattccttg tgtataaggg gacactgtat
     4681 ctgcgtccca caatacaaca aatccgtccc tttacaacaa caaatccgtc ccttcttaac
     4741 aacaaatccg tcccttaatg gcaacaaatc cgtccctttt taaactctac aggccacgga
     4801 ttacgtggcc tgtagacgtc ctaaaaggtt taaaagggaa aaggaagaaa agggtggaaa
     4861 cgcaaaaaac gcaccactac gtggccccgt tggggccgca tttgtgcccc tgaaggggcg
     4921 ggggaggcgt ctgggcaatc cccgttttac cagtccccta tcgccgcctg agagggcgca
     4981 ggaagcgagt aatcagggta tcgaggcgga ttcacccttg gcgtccaacc agcggcacca
     5041 gcggcgcctg agaggggcgc gcccagctgt ctagggcggc ggatttgtcc tactcaggag
     5101 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
     5161 ttttatttga tgcct
//
