LOCUS       40924_27309        5716 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL0C_2-R, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5716)
  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 5716)
  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 5716)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5716)
  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..5716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(581..1381)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1382..1602)
                     /label=URA3 promoter
     rep_origin      complement(2201..3081)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3430..4221)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4222..4326)
                     /label=AmpR promoter
     misc_feature    4395..4434
                     /label=8 cutter set: AscI, SbfI, SwaI, FseI, PmeI
                     /note="8 cutter set: AscI, SbfI, SwaI, FseI, PmeI"
     misc_feature    4435..4470
                     /label=homology region C2*
                     /note="homology region C2*"
     CDS             4806..5108
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     promoter        5276..5462
                     /label=HIS3 promoter
     misc_feature    5463..5512
                     /label=homology region CR*
                     /note="homology region CR*"
     misc_feature    5516..5555
                     /label=8 cutter set: SwaI, AscI, PmeI, SbfI, FseI
                     /note="8 cutter set: SwaI, AscI, PmeI, SbfI, FseI"
     rep_origin      5581..5716
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
       61 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
      121 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
      181 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
      241 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
      301 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      361 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      481 ttcctggcct tttgctggcc ttttgctcat gatataatta aattgaagct ctaatttgtg
      541 agtttagtat acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc
      601 tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc
      661 cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat
      721 ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg
      781 tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc
      841 cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag
      901 tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg
      961 ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg
     1021 cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga
     1081 ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg
     1141 ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat
     1201 gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg
     1261 tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata
     1321 gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca
     1381 tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc
     1441 aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct
     1501 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa agaaaccgaa
     1561 atcaaaaaaa agaataaaaa aaaaatgatg aattgaattg aaaagctagc ttatcgatga
     1621 taagctgtca aagatgagaa ttaattccac ggactataga ctatactaga tactccgtct
     1681 actgtacgat acacttccgc tcaggtcctt gtcctttaac gaggccttac cactcttttg
     1741 ttactctatt gatccagctc agcaaaggca gtgtgatcta agattctatc ttcgcgatgt
     1801 agtaaaacta gctagaccga gaaagagact agaaatgcaa aaggcacttc tacaatggct
     1861 gccatcatta ttatccgatg tgacgctgca gcttctcaat gatattcgaa tacgctttga
     1921 ggagatacag cctaatatcc gacaaactgt tttacagatt tacgatcgta cttgttaccc
     1981 atcattgaat tttgaacatc cgaacctggg agttttccct gaaacagata gtatatttga
     2041 acctgtataa taatatatag tctagcgctt tacggaagac aatgtatgta tttcggttcc
     2101 tggagaaact attgcatcta ttgcataggt aatcttgcac gtcgcatccc cggttcattt
     2161 tctgcgtttc catcttgcac ttcaatagca tatctttgtt aacgaagcat ctgtgcttca
     2221 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     2281 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     2341 ttcatttttg taaaacaaaa atgcaacgcg acgagagcgc taatttttca aacaaagaat
     2401 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     2461 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     2521 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     2581 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     2641 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     2701 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     2761 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     2821 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     2881 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     2941 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     3001 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     3061 tgtggaagcg gtattcgcaa tgggaagctc caccccggtt gataatcaga aaagccccaa
     3121 aaacaggaag attgtataag caaatattta aattgtaatt ctttcctgcg ttatcccctg
     3181 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3241 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gggtctgacg ctcagtggaa
     3301 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3361 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3421 tgacagtcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     3481 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     3541 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     3601 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgtcatg
     3661 ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc
     3721 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     3781 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     3841 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     3901 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     3961 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     4021 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     4081 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     4141 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     4201 caatccatct tgttcaatca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4261 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4321 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgga cggatcgctt gcctgtaact
     4381 tacacgcgcc tcgtggcgcg cccctgcagg atttaaatgg ccggccgttt aaactagtaa
     4441 gttattatcg cactcacgag aagcgtattc aagcttggct tactaaaagc cagataacag
     4501 tatgcgtatt tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga
     4561 agtatgtcaa aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca
     4621 gctatcagtt gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat
     4681 gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat
     4741 ggctgaggtc gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg
     4801 gtgaaatgca gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg
     4861 atgtacagag tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg
     4921 cacgtctgct gtcagataaa gtcccccgtg aactttaccc ggtggtgcat atcggggatg
     4981 aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag
     5041 aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct
     5101 ggggaatata aatgtcaggc tcccttatac acagccagta agcttaattc ccgttttaag
     5161 agcttggtga gcgctaggag tcactgccag gtatcgtttg aacacggcat tagtcaggga
     5221 agtcataaca cagtcctttc ccgcaatttt ctttttctat tactcttggc ctcctctagt
     5281 acactctata tttttttatg cctcggtaat gattttcatt tttttttttc ccctagcgga
     5341 tgactctttt tttttcttag cgattggcat tatcacataa tgaattatac attatataaa
     5401 gtaatgtgat ttcttcgaag aatatactaa aaaatgagca ggcaagataa acgaaggcaa
     5461 agatgacaga gcagaaagcc ctagtaaagc gtattacaaa tgaaaccaag attgaattta
     5521 aatggcgcgc cgtttaaacc ctgcaggggc cggccccgta gaaaagatca aaggatcttc
     5581 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5641 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5701 cagcagagcg cagata
//
