LOCUS       40924_27519        3699 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL2AE-lc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3699)
  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 3699)
  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 3699)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3699)
  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..3699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(57..645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(819..1676)
                     /label=AmpR
                     /note="beta-lactamase"
     misc_feature    complement(1774..2277)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        2538..2806
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        2815..2889
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      2894..2913
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      2965..3154
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    3373..3390
                     /note="I-SceI; cleavage site"
     misc_feature    3391..3440
                     /label=homology region A0*
                     /note="homology region A0*"
     misc_feature    3447..3496
                     /label=homology region F0*
                     /note="homology region F0*"
     misc_feature    3497..3514
                     /label=I-SceI
                     /note="I-SceI"
ORIGIN
        1 tgagcaaaag gccagcaaaa gcccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
       61 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      481 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
      841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      901 ataactacga tacgggagcg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      961 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1021 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1081 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttggcattgc tacaggcatc
     1141 gtggtgtcac tctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1201 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1261 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1321 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1381 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1441 aatagtgtat cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1501 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1561 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1621 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1681 ttcctttttc aatgggtaat aactgatata attaaattga agctctaatt tgtgagttta
     1741 gtatacatgc atttacttat aatacagttg acggatcgct tgcctgtaac ttacacgcgc
     1801 ctcgtatctt ttaatgatgg aataatttgg gaatttactc tgtgtttatt tatttttatg
     1861 ttttgtattt ggattttaga aagtaaataa agaaggtaga agagttacgg aatgaagaaa
     1921 aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat atatatttat
     1981 tagacaagaa aagcagatta aatagatata cattcgatta acgataagta aaatgtaaaa
     2041 tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc ggcattaata
     2101 cctgagagca ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc ctagagtctt
     2161 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt
     2221 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa
     2281 aaggaccacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2341 tttaacgaat agcccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2401 gggttgagtg ttgttccagt ttccaacaag agtccactat taaagaacgt ggactccaac
     2461 gtcaaagggc gaaaaagggt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2521 tcaagttttt tggggtctct ttgaaaagat aatgtatgat tatgctttca ctcatattta
     2581 tacagaaact tgatgttttc tttcgagtat atacaaggtg attacatgta cgtttgaagt
     2641 acaactctag attttgtagt gccctcttgg gctagcggta aaggtgcgca ttttttcaca
     2701 ccctacaatg ttctgttcaa aagattttgg tcaaacgctg tagaagtgaa agttggtgcg
     2761 catgtttcgg cgttcgaaac ttctccgcag tgaaagataa atgatcgcgg ccgcgtttta
     2821 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     2881 gagtcggtgg tgcttttttt gttttttatg tcttcgagtc atgtaattag ttatgtcacg
     2941 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     3001 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     3061 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     3121 aaccttgctt gagaaggttt tgggacgctc gaaggcttta ggtgccgtaa agcagtaaat
     3181 cggaagggta aacggatgcc cccatttaga gcttgacggg gaaagccggc gaacgtggcg
     3241 agaaaggaag ggaagaaagc gaaaggagcg ggggctaggg cggtgggaag tgtaggggtc
     3301 acgctgggcg taaccaccac acccgccgcg cttaatgggg cgctacaggg cgcgtgggga
     3361 tgatccacta gttagggata acagggtaat aaataggaaa gtctggtaac tcaggacact
     3421 gtatctgcta cgctgtttat gaattcaaag tattggtgtg agtaatcgct atctcttcta
     3481 cgactggcta caacaaatta ccctgttatc cctacggccc tgcattaatg aatcggccaa
     3541 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3601 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3661 ttatccacag aatcagggga taacgcagga aagaacatg
//
