LOCUS       40924_27419        5554 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pL1A-hc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5554)
  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 5554)
  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 5554)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5554)
  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..5554
                     /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 A0*
                     /note="homology region A0*"
     CDS             4407..5207
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     misc_feature    5306..5355
                     /label=homology region B0*
                     /note="homology region B0*"
     misc_feature    5356..5373
                     /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 agggtaataa ataggaaagt ctggtaactc aggacactgt
     4381 atctgctacg ctgtttattt aattaaatgt cgaaagctac atataaggaa cgtgctgcta
     4441 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     4501 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     4561 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     4621 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca
     4681 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     4741 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     4801 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     4861 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     4921 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     4981 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg
     5041 cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct gacattatta
     5101 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     5161 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     5221 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     5281 tattacccat tgaaaaagga agagttaagg tttcagtcac actacggata cttttacaac
     5341 gggagcagtt attcaattac cctgttatcc ctacggccct gcattaatga atcggccaac
     5401 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5461 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5521 tatccacaga atcaggggat aacgcaggaa agaa
//
