LOCUS       40924_27449        9038 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL1D-hc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9038)
  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 9038)
  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 9038)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9038)
  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..9038
                     /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"
     rep_origin      complement(2364..3244)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3496..3764
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA
                     gene SNR52"
     misc_RNA        3773..3847
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      3852..3871
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4
                     tRNA gene"
     terminator      3923..4112
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    4331..4348
                     /note="I-SceI; cleavage site"
     misc_feature    4349..4398
                     /label=homology region D0*
                     /note="homology region D0*"
     CDS             4407..8582
                     /label=LYS2
                     /note="alpha-aminoadipate reductase, required for lysine 
                     biosynthesis"
     misc_feature    8790..8839
                     /label=homology region E0*
                     /note="homology region E0*"
     misc_feature    8840..8857
                     /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 gttttatcga tgataagctg tcaaagatga
     1801 gaattaattc cacggactat agactatact agatactccg tctactgtac gatacacttc
     1861 cgctcaggtc cttgtccttt aacgaggcct taccactctt ttgttactct attgatccag
     1921 ctcagcaaag gcagtgtgat ctaagattct atcttcgcga tgtagtaaaa ctagctagac
     1981 cgagaaagag actagaaatg caaaaggcac ttctacaatg gctgccatca ttattatccg
     2041 atgtgacgct gcagcttctc aatgatattc gaatacgctt tgaggagata cagcctaata
     2101 tccgacaaac tgttttacag atttacgatc gtacttgtta cccatcattg aattttgaac
     2161 atccgaacct gggagttttc cctgaaacag atagtatatt tgaacctgta taataatata
     2221 tagtctagcg ctttacggaa gacaatgtat gtatttcggt tcctggagaa actattgcat
     2281 ctattgcata ggtaatcttg cacgtcgcat ccccggttca ttttctgcgt ttccatcttg
     2341 cacttcaata gcatatcttt gttaacgaag catctgtgct tcattttgta gaacaaaaat
     2401 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     2461 aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca
     2521 aaaatgcaac gcgacgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     2581 agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
     2641 gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
     2701 ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
     2761 ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
     2821 ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
     2881 catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
     2941 agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
     3001 tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
     3061 ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
     3121 ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
     3181 tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
     3241 caatgacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3301 taacgaatag cccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3361 gttgagtgtt gttccagttt ccaacaagag tccactatta aagaacgtgg actccaacgt
     3421 caaagggcga aaaagggtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3481 aagttttttg gggtctcttt gaaaagataa tgtatgatta tgctttcact catatttata
     3541 cagaaacttg atgttttctt tcgagtatat acaaggtgat tacatgtacg tttgaagtac
     3601 aactctagat tttgtagtgc cctcttgggc tagcggtaaa ggtgcgcatt ttttcacacc
     3661 ctacaatgtt ctgttcaaaa gattttggtc aaacgctgta gaagtgaaag ttggtgcgca
     3721 tgtttcggcg ttcgaaactt ctccgcagtg aaagataaat gatcgcggcc gcgttttaga
     3781 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     3841 gtcggtggtg ctttttttgt tttttatgtc ttcgagtcat gtaattagtt atgtcacgct
     3901 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     3961 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     4021 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     4081 ccttgcttga gaaggttttg ggacgctcga aggctttagg tgccgtaaag cagtaaatcg
     4141 gaagggtaaa cggatgcccc catttagagc ttgacgggga aagccggcga acgtggcgag
     4201 aaaggaaggg aagaaagcga aaggagcggg ggctagggcg gtgggaagtg taggggtcac
     4261 gctgggcgta accaccacac ccgccgcgct taatggggcg ctacagggcg cgtggggatg
     4321 atccactagt tagggataac agggtaatca cgagcgttga ggcggatttc agattattta
     4381 cagacactat tctacaattt aattaaatga ctaacgaaaa ggtctggata gagaagttgg
     4441 ataatccaac tctttcagtg ttaccacatg actttttacg cccacaacaa gaaccttata
     4501 cgaaacaagc tacatattcg ttacagctac ctcagctcga tgtgcctcat gatagttttt
     4561 ctaacaaata cgctgtcgct ttgagtgtat gggctgcatt gatatataga gtaaccggtg
     4621 acgatgatat tgttctttat attgcgaata acaaaatctt aagattcaat attcaaccaa
     4681 cgtggtcatt taatgagctg tattctacaa ttaacaatga gttgaacaag ctcaattcta
     4741 ttgaggccaa tttttccttt gacgagctag ctgaaaaaat tcaaagttgc caagatctgg
     4801 aaaggacccc tcagttgttc cgtttggcct ttttggaaaa ccaagatttc aaattagacg
     4861 agttcaagca tcatttagtg gactttgctt tgaatttgga taccagtaat aatgcgcatg
     4921 ttttgaactt aatttataac agcttactgt attcgaatga aagagtaacc attgttgcgg
     4981 accaatttac tcaatatttg actgctgcgc taagcgatcc atccaattgc ataactaaaa
     5041 tctctctgat caccgcatca tccaaggata gtttacctga tccaactaag aacttgggct
     5101 ggtgcgattt cgtggggtgt attcacgaca ttttccagga caatgctgaa gccttcccag
     5161 agagaacctg tgttgtggag actccaacac taaattccga caagtcccgt tctttcactt
     5221 atcgcgacat caaccgcact tctaacatag ttgcccatta tttgattaaa acaggtatca
     5281 aaagaggtga tgtagtgatg atctattctt ctaggggtgt ggatttgatg gtatgtgtga
     5341 tgggtgtctt gaaagccggc gcaacctttt cagttatcga ccctgcatat cccccagcca
     5401 gacaaaccat ttacttaggt gttgctaaac cacgtgggtt gattgttatt agagctgctg
     5461 gacaattgga tcaactagta gaagattaca tcaatgatga attggagatt gtttcaagaa
     5521 tcaattccat cgctattcaa gaaaatggta ccattgaagg tggcaaattg gacaatggcg
     5581 aggatgtttt ggctccatat gatcactaca aagacaccag aacaggtgtt gtagttggac
     5641 cagattccaa cccaacccta tctttcacat ctggttccga aggtattcct aagggtgttc
     5701 ttggtagaca tttttccttg gcttattatt tcaattggat gtccaaaagg ttcaacttaa
     5761 cagaaaatga taaattcaca atgctgagcg gtattgcaca tgatccaatt caaagagata
     5821 tgtttacacc attattttta ggtgcccaat tgtatgtccc tactcaagat gatattggta
     5881 caccgggccg tttagcggaa tggatgagta agtatggttg cacagttacc catttaacac
     5941 ctgccatggg tcaattactt actgcccaag ctactacacc attccctaag ttacatcatg
     6001 cgttctttgt gggtgacatt ttaacaaaac gtgattgtct gaggttacaa accttggcag
     6061 aaaattgccg tattgttaat atgtacggta ccactgaaac acagcgtgca gtttcttatt
     6121 tcgaagttaa atcaaaaaat gacgatccaa actttttgaa aaaattgaaa gatgtcatgc
     6181 ctgctggtaa aggtatgttg aacgttcagc tactagttgt taacaggaac gatcgtactc
     6241 aaatatgtgg tattggcgaa ataggtgaga tttatgttcg tgcaggtggt ttggccgaag
     6301 gttatagagg attaccagaa ttgaataaag aaaaatttgt gaacaactgg tttgttgaaa
     6361 aagatcactg gaattatttg gataaggata atggtgaacc ttggagacaa ttctggttag
     6421 gtccaagaga tagattgtac agaacgggtg atttaggtcg ttatctacca aacggtgact
     6481 gtgaatgttg cggtagggct gatgatcaag ttaaaattcg tgggttcaga atcgaattag
     6541 gagaaataga tacgcacatt tcccaacatc cattggtaag agaaaacatt actttagttc
     6601 gcaaaaatgc cgacaatgag ccaacattga tcacatttat ggtcccaaga tttgacaagc
     6661 cagatgactt gtctaagttc caaagtgatg ttccaaagga ggttgaaact gaccctatag
     6721 ttaagggctt aatcggttac catcttttat ccaaggacat caggactttc ttaaagaaaa
     6781 gattggctag ctatgctatg ccttccttga ttgtggttat ggataaacta ccattgaatc
     6841 caaatggtaa agttgataag cctaaacttc aattcccaac tcccaagcaa ttaaatttgg
     6901 tagctgaaaa tacagtttct gaaactgacg actctcagtt taccaatgtt gagcgcgagg
     6961 ttagagactt atggttaagt atattaccta ccaagccagc atctgtatca ccagatgatt
     7021 cgtttttcga tttaggtggt cattctatct tggctaccaa aatgattttt accttaaaga
     7081 aaaagctgca agttgattta ccattgggca caattttcaa gtatccaacg ataaaggcct
     7141 ttgccgcgga aattgacaga attaaatcat cgggtggatc atctcaaggt gaggtcgtcg
     7201 aaaatgtcac tgcaaattat gcggaagacg ccaagaaatt ggttgagacg ctaccaagtt
     7261 cgtacccctc tcgagaatat tttgttgaac ctaatagtgc cgaaggaaaa acaacaatta
     7321 atgtgtttgt taccggtgtc acaggatttc tgggctccta catccttgca gatttgttag
     7381 gacgttctcc aaagaactac agtttcaaag tgtttgccca cgtcagggcc aaggatgaag
     7441 aagctgcatt tgcaagatta caaaaggcag gtatcaccta tggtacttgg aacgaaaaat
     7501 ttgcctcaaa tattaaagtt gtattaggcg atttatctaa aagccaattt ggtctttcag
     7561 atgagaagtg gatggatttg gcaaacacag ttgatataat tatccataat ggtgcgttag
     7621 ttcactgggt ttatccatat gccaaattga gggatccaaa tgttatttca actatcaatg
     7681 ttatgagctt agccgccgtc ggcaagccaa agttctttga ctttgtttcc tccacttcta
     7741 ctcttgacac tgaatactac tttaatttgt cagataaact tgttagcgaa gggaagccag
     7801 gcattttaga atcagacgat ttaatgaact ctgcaagcgg gctcactggt ggatatggtc
     7861 agtccaaatg ggctgctgag tacatcatta gacgtgcagg tgaaaggggc ctacgtgggt
     7921 gtattgtcag accaggttac gtaacaggtg cctctgccaa tggttcttca aacacagatg
     7981 atttcttatt gagatttttg aaaggttcag tccaattagg taagattcca gatatcgaaa
     8041 attccgtgaa tatggttcca gtagatcatg ttgctcgtgt tgttgttgct acgtctttga
     8101 atcctcccaa agaaaatgaa ttggccgttg ctcaagtaac gggtcaccca agaatattat
     8161 tcaaagacta cttgtatact ttacacgatt atggttacga tgtcgaaatc gaaagctatt
     8221 ctaaatggaa gaaatcattg gaggcgtctg ttattgacag gaatgaagaa aatgcgttgt
     8281 atcctttgct acacatggtc ttagacaact tacctgaaag taccaaagct ccggaactag
     8341 acgataggaa cgccgtggca tctttaaaga aagacaccgc atggacaggt gttgattggt
     8401 ctaatggaat aggtgttact ccagaagagg ttggtatata tattgcattt ttaaacaagg
     8461 ttggattttt acctccacca actcataatg acaaacttcc actgccaagt atagaactaa
     8521 ctcaagcgca aataagtcta gttgcttcag gtgctggtgc tcgtggaagc tccgcagcag
     8581 cttaaggttg agcattacgt atgatatgtc catgtacaat aattaaatat gaattaggag
     8641 aaagacttag cttcttttcg ggtgatgtca cttaaaaact ccgagaataa tatataataa
     8701 gagaataaaa tattagttat tgaataagaa ctgtaaatca gggagatctg catgtctact
     8761 aaactcacaa attagagctt caatttaatt ctcttacaat aaaatagaat actctgcgaa
     8821 gggcgtgaca ctgctgttta ttaccctgtt atccctacgg ccctgcatta atgaatcggc
     8881 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     8941 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     9001 cggttatcca cagaatcagg ggataacgca ggaaagaa
//
