LOCUS       62056_1895        4817 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector p191 LI-TA, complete sequence.
ACCESSION   MK495750
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4817)
  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 4817)
  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 4817)
  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..4817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    59..183
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        220..250
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             304..960
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1305..1607
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1651..1775)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      1885..1979
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        2052..2143
                     /label=AmpR promoter
     CDS             2144..3001
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3008..3047
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            3194..3302
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      3338..3926
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4112..4252)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4357..4545)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      complement(4725..4811)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccaaaat taaaatggac
       61 aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa
      121 ttagattttg cataaaaaac agactacata atactgtaaa acacaacata tccagtcact
      181 atggcggccc ctcgaggggc cgcattaggc accccaggct ttacacttta tgcttccggc
      241 tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct
      301 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
      361 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
      421 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
      481 attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac
      541 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
      601 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
      661 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
      721 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
      781 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
      841 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
      901 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
      961 taatctagag gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat
     1021 ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg
     1081 ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat
     1141 atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct
     1201 gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat
     1261 tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt
     1321 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     1381 acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     1441 tcccccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     1501 ccgatatggc cagtgtgccg gtttccgtta tcggggaaga agtggctgat ctcagccacc
     1561 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct
     1621 cccttataca cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag
     1681 tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt
     1741 tacgtttctc gttcagcttt cttgtacaaa gtggttccat cgaaattttg ggagctcggt
     1801 acccggggat cctctagagt cgacctgcag gcatgcaagc ttgcggccgc gtcgtgactg
     1861 ggaaaaccct ggcgactagt cttggactcc tgttgataga tccagtaatg acctcagaac
     1921 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
     1981 caggggtccc caataattac gatttaaatt agtagcccgc ctaatgagcg ggcttttttt
     2041 taattcccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2101 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagca ttcagcattt
     2161 tcgtgtggcg ctgattccgt tttttgcggc gttttgcctg ccggtgtttg cgcatccgga
     2221 aaccctggtg aaagtgaaag atgcggaaga tcaactgggt gcgcgcgtgg gctatattga
     2281 actggatctg aacagcggca aaattctgga atcttttcgt ccggaagaac gttttccgat
     2341 gatgagcacc tttaaagtgc tgctgtgcgg tgcggttctg agccgtgtgg atgcgggcca
     2401 ggaacaactg ggccgtcgta ttcattatag ccagaacgat ctggtggaat atagcccggt
     2461 gaccgaaaaa catctgaccg atggcatgac cgtgcgtgaa ctgtgcagcg cggcgattac
     2521 catgagcgat aacaccgcgg cgaacctgct gctgacgacc attggcggtc cgaaagaact
     2581 gaccgcgttt ctgcataaca tgggcgatca tgtgacccgt ctggatcgtt gggaaccgga
     2641 actgaacgaa gcgattccga acgatgaacg tgataccacc atgccggcag caatggcgac
     2701 caccctgcgt aaactgctga cgggtgagct gctgaccctg gcaagccgcc agcaactgat
     2761 tgattggatg gaagcggata aagtggcggg tccgctgctg cgtagcgcgc tgccggctgg
     2821 ctggtttatt gcggataaaa gcggtgcggg cgaacgtggc agccgtggca ttattgcggc
     2881 gctgggcccg gatggtaaac cgagccgtat tgtggtgatt tataccaccg gcagccaggc
     2941 gacgatggat gaacgtaacc gtcagattgc ggaaattggc gcgagcctga ttaaacattg
     3001 gtaaaccgat acaattaaag gctccttttg gagccttttt ttttggacga cccttgtcct
     3061 tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct
     3121 ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc
     3181 aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct
     3241 tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc
     3301 cgtaggccgg ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3361 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3421 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3481 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3541 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3601 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3661 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3721 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3781 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3841 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3901 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3961 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4021 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4081 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4141 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     4201 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     4261 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4321 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4381 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgcagcgatt cacagatgtc
     4441 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     4501 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     4561 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     4621 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     4681 ggtatggatg cggcgggggc gcgcccagct gtctagggcg gcggatttgt cctactcagg
     4741 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     4801 cgttttattt gatgcct
//
