LOCUS       40924_27374        5396 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL0E_0-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5396)
  AUTHORS   Hochrein L, Machens F, Gremmels J, Schulz K, Messerschmidt K, 
            Mueller-Roeber B.
  TITLE     AssemblX: a user-friendly toolkit for rapid and reliable multi-gene 
            assemblies
  JOURNAL   Nucleic Acids Res. (2017) In press
  PUBMED    28130422
REFERENCE   2  (bases 1 to 5396)
  AUTHORS   Hochrein L, Machens F, Gremmels J, Schulz K, Messerschmidt K, 
            Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-2016) Department of Molecular Biology - Cell2Fab 
            research unit, University of Potsdam, Karl-Liebknecht-Str. 24-25, 
            Potsdam, Brandenburg 14476, Germany
REFERENCE   3  (bases 1 to 5396)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5396)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-NOV-2016) Department of Molecular Biology - Cell2Fab research 
            unit, University of Potsdam, Karl-Liebknecht-Str. 24-25, Potsdam, 
            Brandenburg 14476, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(557..1357)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1358..1578)
                     /label=URA3 promoter
     rep_origin      complement(2177..3057)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3406..4197)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4198..4302)
                     /label=AmpR promoter
     misc_feature    4371..4410
                     /label=8 cutter set: AscI, SbfI, SwaI, FseI, PmeI
                     /note="8 cutter set: AscI, SbfI, SwaI, FseI, PmeI"
     misc_feature    4411..4460
                     /label=homology region E0*
                     /note="homology region E0*"
     CDS             4796..5098
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    5136..5171
                     /label=homology region E1*
                     /note="homology region E1*"
     misc_feature    5172..5211
                     /label=8 cutter set: SwaI, AscI, PmeI, SbfI, FseI
                     /note="8 cutter set: SwaI, AscI, PmeI, SbfI, FseI"
     rep_origin      5237..5396
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
       61 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
      121 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
      181 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
      241 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
      301 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
      361 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
      421 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      481 gctcatgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
      541 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
      601 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
      661 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
      721 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
      781 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
      841 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
      901 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
      961 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
     1021 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
     1081 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
     1141 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
     1201 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     1261 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     1321 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg
     1381 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
     1441 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
     1501 agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa
     1561 atgatgaatt gaattgaaaa gctagcttat cgatgataag ctgtcaaaga tgagaattaa
     1621 ttccacggac tatagactat actagatact ccgtctactg tacgatacac ttccgctcag
     1681 gtccttgtcc tttaacgagg ccttaccact cttttgttac tctattgatc cagctcagca
     1741 aaggcagtgt gatctaagat tctatcttcg cgatgtagta aaactagcta gaccgagaaa
     1801 gagactagaa atgcaaaagg cacttctaca atggctgcca tcattattat ccgatgtgac
     1861 gctgcagctt ctcaatgata ttcgaatacg ctttgaggag atacagccta atatccgaca
     1921 aactgtttta cagatttacg atcgtacttg ttacccatca ttgaattttg aacatccgaa
     1981 cctgggagtt ttccctgaaa cagatagtat atttgaacct gtataataat atatagtcta
     2041 gcgctttacg gaagacaatg tatgtatttc ggttcctgga gaaactattg catctattgc
     2101 ataggtaatc ttgcacgtcg catccccggt tcattttctg cgtttccatc ttgcacttca
     2161 atagcatatc tttgttaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     2221 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2281 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     2341 aacgcgacga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     2401 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     2461 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     2521 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     2581 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     2641 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     2701 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     2761 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     2821 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     2881 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     2941 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     3001 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatggg
     3061 aagctccacc ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa
     3121 tatttaaatt gtaattcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3181 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3241 gcgaggaagc ggaagagggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3301 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3361 taaatcaatc taaagtatat atgagtaaac ttggtctgac agtcagaaga actcgtcaag
     3421 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     3481 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     3541 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     3601 ttccaccatg atattcggca agcaggcatc gtcatgggtc acgacgagat cctcgccgtc
     3661 gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     3721 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     3781 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     3841 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     3901 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     3961 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag
     4021 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     4081 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     4141 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatact
     4201 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4261 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4321 gccacctgac gtcggacgga tcgcttgcct gtaacttaca cgcgcctcgt ggcgcgcccc
     4381 tgcaggattt aaatggccgg ccgtttaaac tctcttacaa taaaatagaa tactctgcga
     4441 agggcgtgac actgctgttt aagcttggct tactaaaagc cagataacag tatgcgtatt
     4501 tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa
     4561 aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt
     4621 gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa
     4681 gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc
     4741 gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca
     4801 gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag
     4861 tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct
     4921 gtcagataaa gtcccccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg
     4981 catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga
     5041 tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata
     5101 aatgtcaggc tcccttatac acagccagta agcttagttt atcaatccgc tacaggacgg
     5161 tatttcgtta tatttaaatg gcgcgccgtt taaaccctgc aggggccggc cccgtagaaa
     5221 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5281 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5341 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtag
//
