LOCUS       40924_27434        4897 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL1B-lc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4897)
  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 4897)
  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 4897)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4897)
  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..4897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(61..649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(823..1680)
                     /label=AmpR
                     /note="beta-lactamase"
     misc_feature    complement(1778..2281)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        2542..2810
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        2819..2893
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      2898..2917
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      2969..3158
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    3377..3394
                     /note="I-SceI; cleavage site"
     misc_feature    3395..3444
                     /label=homology region B0*
                     /note="homology region B0*"
     CDS             3453..4544
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     misc_feature    4649..4698
                     /label=homology region C0*
                     /note="homology region C0*"
     misc_feature    4699..4716
                     /note="I-SceI; cleavage site"
ORIGIN
        1 catgtgagca aaaggccagc aaaagcccag gaaccgtaaa aaggccgcgt tgctggcgtt
       61 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      121 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      181 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      241 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      301 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      361 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      421 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      481 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
      541 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      601 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      661 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      721 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      781 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      841 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      901 gtagataact acgatacggg agcgcttacc atctggcccc agtgctgcaa tgataccgcg
      961 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1021 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1081 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttggca ttgctacagg
     1141 catcgtggtg tcactctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1201 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1261 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1321 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1381 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1441 ggataatagt gtatcacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1501 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1561 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1621 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1681 actcttcctt tttcaatggg taataactga tataattaaa ttgaagctct aatttgtgag
     1741 tttagtatac atgcatttac ttataataca gttgacggat cgcttgcctg taacttacac
     1801 gcgcctcgta tcttttaatg atggaataat ttgggaattt actctgtgtt tatttatttt
     1861 tatgttttgt atttggattt tagaaagtaa ataaagaagg tagaagagtt acggaatgaa
     1921 gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa agcgtacttt acatatatat
     1981 ttattagaca agaaaagcag attaaataga tatacattcg attaacgata agtaaaatgt
     2041 aaaatcacag gattttcgtg tgtggtcttc tacacagaca agatgaaaca attcggcatt
     2101 aatacctgag agcaggaaga gcaagataaa aggtagtatt tgttggcgat ccccctagag
     2161 tcttttacat cttcggaaaa caaaaactat tttttcttta atttcttttt ttactttcta
     2221 tttttaattt atatatttat attaaaaaat ttaaattata attattttta tagcacgtga
     2281 tgaaaaggac cacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2341 attttttaac gaatagcccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2401 gatagggttg agtgttgttc cagtttccaa caagagtcca ctattaaaga acgtggactc
     2461 caacgtcaaa gggcgaaaaa gggtctatca gggcgatggc ccactacgtg aaccatcacc
     2521 ctaatcaagt tttttggggt ctctttgaaa agataatgta tgattatgct ttcactcata
     2581 tttatacaga aacttgatgt tttctttcga gtatatacaa ggtgattaca tgtacgtttg
     2641 aagtacaact ctagattttg tagtgccctc ttgggctagc ggtaaaggtg cgcatttttt
     2701 cacaccctac aatgttctgt tcaaaagatt ttggtcaaac gctgtagaag tgaaagttgg
     2761 tgcgcatgtt tcggcgttcg aaacttctcc gcagtgaaag ataaatgatc gcggccgcgt
     2821 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     2881 caccgagtcg gtggtgcttt ttttgttttt tatgtcttcg agtcatgtaa ttagttatgt
     2941 cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac
     3001 aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat
     3061 ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac
     3121 tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaggtgcc gtaaagcagt
     3181 aaatcggaag ggtaaacgga tgcccccatt tagagcttga cggggaaagc cggcgaacgt
     3241 ggcgagaaag gaagggaaga aagcgaaagg agcgggggct agggcggtgg gaagtgtagg
     3301 ggtcacgctg ggcgtaacca ccacacccgc cgcgcttaat ggggcgctac agggcgcgtg
     3361 gggatgatcc actagttagg gataacaggg taattaaggt ttcagtcaca ctacggatac
     3421 ttttacaacg ggagcagtta ttcattaatt aaatgtctgc ccctaagaag atcgtcgttt
     3481 tgccaggtga ccacgttggt caagaaatca cagccgaagc cattaaggtt cttaaagcta
     3541 tttctgatgt tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg
     3601 ctatcgatgc tacaggtgtc ccacttccag atgaggcgct ggaagcctcc aagaaggttg
     3661 atgccgtttt gttaggtgct gtgggtggtc ctaaatgggg tacaggtagt gttagacctg
     3721 aacaaggttt actaaaaatc cgtaaagaac ttcaattgta cgccaactta agaccatgta
     3781 actttgcatc cgactctctt ttagacttat ctccaatcaa gccacaattt gctaaaggta
     3841 ctgacttcgt tgttgtcaga gaattagtgg gaggtattta ctttggtaag agaaaggaag
     3901 acgatggtga tggtgtcgct tgggatagtg aacaatacac cgttccagaa gtgcaaagaa
     3961 tcacaagaat ggccgctttc atggccctac aacatgagcc accattgcct atttggtcct
     4021 tggataaagc taatgttttg gcctcttcaa gattatggag aaaaactgtg gaggaaacca
     4081 tcaagaacga atttcctaca ttgaaggttc aacatcaatt gattgattct gccgccatga
     4141 tcctagttaa gaacccaacc cacctaaatg gtattataat caccagcaac atgtttggtg
     4201 atatcatctc cgatgaagcc tccgttatcc caggttcctt gggtttgttg ccatctgcgt
     4261 ccttggcctc tttgccagac aagaacaccg catttggttt gtacgaacca tgccacggtt
     4321 ctgctccaga tttgccaaag aataaggttg accctatcgc cactatcttg tctgctgcaa
     4381 tgatgttgaa attgtcattg aacttgcctg aagaaggtaa ggccattgaa gatgcagtta
     4441 aaaaggtttt ggatgcaggt atcagaactg gtgatttagg tggttccaac agtaccaccg
     4501 aagtcggtga tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt
     4561 ttttatgata tttgtacata aactttataa atgaaattca taatagaaac gacacgaaat
     4621 tacaaaatgg aatatgttca tagggtagta ttgcgtgtga tacttcagga gaaccgttat
     4681 tccgtaaaac tacagtcaat taccctgtta tccctacggc cctgcattaa tgaatcggcc
     4741 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4801 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4861 ggttatccac agaatcaggg gataacgcag gaaagaa
//
