LOCUS       40924_27334        5708 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL0D_0-R, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5708)
  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 5708)
  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 5708)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5708)
  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..5708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(561..1361)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1362..1582)
                     /label=URA3 promoter
     rep_origin      complement(2181..3061)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3410..4201)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4202..4306)
                     /label=AmpR promoter
     misc_feature    4375..4414
                     /label=8 cutter set: AscI, SbfI, SwaI, FseI, PmeI
                     /note="8 cutter set: AscI, SbfI, SwaI, FseI, PmeI"
     misc_feature    4415..4464
                     /label=homology region D0*
                     /note="homology region D0*"
     CDS             4800..5102
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     promoter        5194..5433
                     /label=LYS2 promoter
     misc_feature    5434..5484
                     /label=homology region DR*
                     /note="homology region DR*"
     misc_feature    5488..5527
                     /label=8 cutter set: SwaI, AscI, PmeI, SbfI, FseI
                     /note="8 cutter set: SwaI, AscI, PmeI, SbfI, FseI"
     rep_origin      5553..5708
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
       61 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      121 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      181 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      241 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      301 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      361 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      421 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      481 ttttgctcat gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt
      541 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
      601 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
      661 ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga
      721 cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
      781 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
      841 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
      901 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
      961 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
     1021 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
     1081 aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
     1141 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
     1201 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
     1261 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
     1321 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
     1381 caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
     1441 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     1501 tcggagatta ccgaatcaaa aaaatttcaa agaaaccgaa atcaaaaaaa agaataaaaa
     1561 aaaaatgatg aattgaattg aaaagctagc ttatcgatga taagctgtca aagatgagaa
     1621 ttaattccac ggactataga ctatactaga tactccgtct actgtacgat acacttccgc
     1681 tcaggtcctt gtcctttaac gaggccttac cactcttttg ttactctatt gatccagctc
     1741 agcaaaggca gtgtgatcta agattctatc ttcgcgatgt agtaaaacta gctagaccga
     1801 gaaagagact agaaatgcaa aaggcacttc tacaatggct gccatcatta ttatccgatg
     1861 tgacgctgca gcttctcaat gatattcgaa tacgctttga ggagatacag cctaatatcc
     1921 gacaaactgt tttacagatt tacgatcgta cttgttaccc atcattgaat tttgaacatc
     1981 cgaacctggg agttttccct gaaacagata gtatatttga acctgtataa taatatatag
     2041 tctagcgctt tacggaagac aatgtatgta tttcggttcc tggagaaact attgcatcta
     2101 ttgcataggt aatcttgcac gtcgcatccc cggttcattt tctgcgtttc catcttgcac
     2161 ttcaatagca tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca
     2221 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     2281 gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa
     2341 atgcaacgcg acgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     2401 acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt
     2461 ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt
     2521 tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta
     2581 aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca
     2641 cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat
     2701 ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc
     2761 gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat
     2821 actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc
     2881 ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt
     2941 cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag
     3001 cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa
     3061 tgggaagctc caccccggtt gataatcaga aaagccccaa aaacaggaag attgtataag
     3121 caaatattta aattgtaatt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3181 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3241 gtgagcgagg aagcggaaga gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3301 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3361 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagtcag aagaactcgt
     3421 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     3481 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     3541 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     3601 cattttccac catgatattc ggcaagcagg catcgtcatg ggtcacgacg agatcctcgc
     3661 cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     3721 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
     3781 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     3841 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     3901 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     3961 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt
     4021 gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     4081 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     4141 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
     4201 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4261 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4321 aagtgccacc tgacgtcgga cggatcgctt gcctgtaact tacacgcgcc tcgtggcgcg
     4381 cccctgcagg atttaaatgg ccggccgttt aaaccacgag cgttgaggcg gatttcagat
     4441 tatttacaga cactattcta caataagctt ggcttactaa aagccagata acagtatgcg
     4501 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
     4561 tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc
     4621 agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa
     4681 tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga
     4741 ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa
     4801 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     4861 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc
     4921 tgctgtcaga taaagtcccc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     4981 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     5041 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     5101 tataaatgtc aggctccctt atacacagcc agtaagcttt agaggcatcg cacagtttta
     5161 gcgaggaaaa ctcttcaata gttttgccag cggaattcca cttgcaatta cataaaaaat
     5221 tccggcggtt tttcgcgtgt gactcaatgt cgaaatacct gcctaatgaa catgaacatc
     5281 gcccaaatgt atttgaagac ccgctgggag aagttcaaga tatataagta acaagcagcc
     5341 aatagtataa aaaaaaatct gagtttatta cctttcctgg aatttcagtg aaaaactgct
     5401 aattatagag agatatcaca gagttactca ctaatgacta acgaaaaggt ctggatagag
     5461 aagttggata atccaactct ttcatgaatt taaatggcgc gccgtttaaa ccctgcaggg
     5521 gccggccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5701 cttctagt
//
