LOCUS       40924_27454        8084 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL1D-lc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8084)
  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 8084)
  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 8084)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8084)
  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..8084
                     /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 D0*
                     /note="homology region D0*"
     CDS             3453..7628
                     /label=LYS2
                     /note="alpha-aminoadipate reductase, required for lysine 
                     biosynthesis"
     misc_feature    7836..7885
                     /label=homology region E0*
                     /note="homology region E0*"
     misc_feature    7886..7903
                     /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 taatcacgag cgttgaggcg gatttcagat
     3421 tatttacaga cactattcta caatttaatt aaatgactaa cgaaaaggtc tggatagaga
     3481 agttggataa tccaactctt tcagtgttac cacatgactt tttacgccca caacaagaac
     3541 cttatacgaa acaagctaca tattcgttac agctacctca gctcgatgtg cctcatgata
     3601 gtttttctaa caaatacgct gtcgctttga gtgtatgggc tgcattgata tatagagtaa
     3661 ccggtgacga tgatattgtt ctttatattg cgaataacaa aatcttaaga ttcaatattc
     3721 aaccaacgtg gtcatttaat gagctgtatt ctacaattaa caatgagttg aacaagctca
     3781 attctattga ggccaatttt tcctttgacg agctagctga aaaaattcaa agttgccaag
     3841 atctggaaag gacccctcag ttgttccgtt tggccttttt ggaaaaccaa gatttcaaat
     3901 tagacgagtt caagcatcat ttagtggact ttgctttgaa tttggatacc agtaataatg
     3961 cgcatgtttt gaacttaatt tataacagct tactgtattc gaatgaaaga gtaaccattg
     4021 ttgcggacca atttactcaa tatttgactg ctgcgctaag cgatccatcc aattgcataa
     4081 ctaaaatctc tctgatcacc gcatcatcca aggatagttt acctgatcca actaagaact
     4141 tgggctggtg cgatttcgtg gggtgtattc acgacatttt ccaggacaat gctgaagcct
     4201 tcccagagag aacctgtgtt gtggagactc caacactaaa ttccgacaag tcccgttctt
     4261 tcacttatcg cgacatcaac cgcacttcta acatagttgc ccattatttg attaaaacag
     4321 gtatcaaaag aggtgatgta gtgatgatct attcttctag gggtgtggat ttgatggtat
     4381 gtgtgatggg tgtcttgaaa gccggcgcaa ccttttcagt tatcgaccct gcatatcccc
     4441 cagccagaca aaccatttac ttaggtgttg ctaaaccacg tgggttgatt gttattagag
     4501 ctgctggaca attggatcaa ctagtagaag attacatcaa tgatgaattg gagattgttt
     4561 caagaatcaa ttccatcgct attcaagaaa atggtaccat tgaaggtggc aaattggaca
     4621 atggcgagga tgttttggct ccatatgatc actacaaaga caccagaaca ggtgttgtag
     4681 ttggaccaga ttccaaccca accctatctt tcacatctgg ttccgaaggt attcctaagg
     4741 gtgttcttgg tagacatttt tccttggctt attatttcaa ttggatgtcc aaaaggttca
     4801 acttaacaga aaatgataaa ttcacaatgc tgagcggtat tgcacatgat ccaattcaaa
     4861 gagatatgtt tacaccatta tttttaggtg cccaattgta tgtccctact caagatgata
     4921 ttggtacacc gggccgttta gcggaatgga tgagtaagta tggttgcaca gttacccatt
     4981 taacacctgc catgggtcaa ttacttactg cccaagctac tacaccattc cctaagttac
     5041 atcatgcgtt ctttgtgggt gacattttaa caaaacgtga ttgtctgagg ttacaaacct
     5101 tggcagaaaa ttgccgtatt gttaatatgt acggtaccac tgaaacacag cgtgcagttt
     5161 cttatttcga agttaaatca aaaaatgacg atccaaactt tttgaaaaaa ttgaaagatg
     5221 tcatgcctgc tggtaaaggt atgttgaacg ttcagctact agttgttaac aggaacgatc
     5281 gtactcaaat atgtggtatt ggcgaaatag gtgagattta tgttcgtgca ggtggtttgg
     5341 ccgaaggtta tagaggatta ccagaattga ataaagaaaa atttgtgaac aactggtttg
     5401 ttgaaaaaga tcactggaat tatttggata aggataatgg tgaaccttgg agacaattct
     5461 ggttaggtcc aagagataga ttgtacagaa cgggtgattt aggtcgttat ctaccaaacg
     5521 gtgactgtga atgttgcggt agggctgatg atcaagttaa aattcgtggg ttcagaatcg
     5581 aattaggaga aatagatacg cacatttccc aacatccatt ggtaagagaa aacattactt
     5641 tagttcgcaa aaatgccgac aatgagccaa cattgatcac atttatggtc ccaagatttg
     5701 acaagccaga tgacttgtct aagttccaaa gtgatgttcc aaaggaggtt gaaactgacc
     5761 ctatagttaa gggcttaatc ggttaccatc ttttatccaa ggacatcagg actttcttaa
     5821 agaaaagatt ggctagctat gctatgcctt ccttgattgt ggttatggat aaactaccat
     5881 tgaatccaaa tggtaaagtt gataagccta aacttcaatt cccaactccc aagcaattaa
     5941 atttggtagc tgaaaataca gtttctgaaa ctgacgactc tcagtttacc aatgttgagc
     6001 gcgaggttag agacttatgg ttaagtatat tacctaccaa gccagcatct gtatcaccag
     6061 atgattcgtt tttcgattta ggtggtcatt ctatcttggc taccaaaatg atttttacct
     6121 taaagaaaaa gctgcaagtt gatttaccat tgggcacaat tttcaagtat ccaacgataa
     6181 aggcctttgc cgcggaaatt gacagaatta aatcatcggg tggatcatct caaggtgagg
     6241 tcgtcgaaaa tgtcactgca aattatgcgg aagacgccaa gaaattggtt gagacgctac
     6301 caagttcgta cccctctcga gaatattttg ttgaacctaa tagtgccgaa ggaaaaacaa
     6361 caattaatgt gtttgttacc ggtgtcacag gatttctggg ctcctacatc cttgcagatt
     6421 tgttaggacg ttctccaaag aactacagtt tcaaagtgtt tgcccacgtc agggccaagg
     6481 atgaagaagc tgcatttgca agattacaaa aggcaggtat cacctatggt acttggaacg
     6541 aaaaatttgc ctcaaatatt aaagttgtat taggcgattt atctaaaagc caatttggtc
     6601 tttcagatga gaagtggatg gatttggcaa acacagttga tataattatc cataatggtg
     6661 cgttagttca ctgggtttat ccatatgcca aattgaggga tccaaatgtt atttcaacta
     6721 tcaatgttat gagcttagcc gccgtcggca agccaaagtt ctttgacttt gtttcctcca
     6781 cttctactct tgacactgaa tactacttta atttgtcaga taaacttgtt agcgaaggga
     6841 agccaggcat tttagaatca gacgatttaa tgaactctgc aagcgggctc actggtggat
     6901 atggtcagtc caaatgggct gctgagtaca tcattagacg tgcaggtgaa aggggcctac
     6961 gtgggtgtat tgtcagacca ggttacgtaa caggtgcctc tgccaatggt tcttcaaaca
     7021 cagatgattt cttattgaga tttttgaaag gttcagtcca attaggtaag attccagata
     7081 tcgaaaattc cgtgaatatg gttccagtag atcatgttgc tcgtgttgtt gttgctacgt
     7141 ctttgaatcc tcccaaagaa aatgaattgg ccgttgctca agtaacgggt cacccaagaa
     7201 tattattcaa agactacttg tatactttac acgattatgg ttacgatgtc gaaatcgaaa
     7261 gctattctaa atggaagaaa tcattggagg cgtctgttat tgacaggaat gaagaaaatg
     7321 cgttgtatcc tttgctacac atggtcttag acaacttacc tgaaagtacc aaagctccgg
     7381 aactagacga taggaacgcc gtggcatctt taaagaaaga caccgcatgg acaggtgttg
     7441 attggtctaa tggaataggt gttactccag aagaggttgg tatatatatt gcatttttaa
     7501 acaaggttgg atttttacct ccaccaactc ataatgacaa acttccactg ccaagtatag
     7561 aactaactca agcgcaaata agtctagttg cttcaggtgc tggtgctcgt ggaagctccg
     7621 cagcagctta aggttgagca ttacgtatga tatgtccatg tacaataatt aaatatgaat
     7681 taggagaaag acttagcttc ttttcgggtg atgtcactta aaaactccga gaataatata
     7741 taataagaga ataaaatatt agttattgaa taagaactgt aaatcaggga gatctgcatg
     7801 tctactaaac tcacaaatta gagcttcaat ttaattctct tacaataaaa tagaatactc
     7861 tgcgaagggc gtgacactgc tgtttattac cctgttatcc ctacggccct gcattaatga
     7921 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7981 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     8041 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaa
//
