LOCUS       40924_34256        6720 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPC_020_GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6720)
  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 6720)
  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 6720)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6720)
  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..6720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(480..1280)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1281..1501)
                     /label=URA3 promoter
     rep_origin      complement(2100..2980)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3329..4120)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4121..4225)
                     /label=AmpR promoter
     promoter        4513..5217
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             5232..5945
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5957..6122
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        6143..6363
                     /label=URA3 promoter
     rep_origin      6484..6720
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
       61 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcatg atataattaa
      421 attgaagctc taatttgtga gtttagtata catgcattta cttataatac agttttttag
      481 ttttgctggc cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc
      541 tctaccttag catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat
      601 cctgtagaga ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca
      661 tctaaaccca caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg
      721 tctctttgag caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta
      781 cccttagtat attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa
      841 aggcctctag gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct
      901 gggcccacca caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc
      961 gcagagtact gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt
     1021 aaaaaattgt acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa
     1081 tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact
     1141 aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt
     1201 tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc
     1261 ttatatgtag ctttcgacat gatttatctt cgtttcctgc aggtttttgt tctgtgcagt
     1321 tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc
     1381 aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa
     1441 aaatttcaaa gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaattga
     1501 aaagctagct tatcgatgat aagctgtcaa agatgagaat taattccacg gactatagac
     1561 tatactagat actccgtcta ctgtacgata cacttccgct caggtccttg tcctttaacg
     1621 aggccttacc actcttttgt tactctattg atccagctca gcaaaggcag tgtgatctaa
     1681 gattctatct tcgcgatgta gtaaaactag ctagaccgag aaagagacta gaaatgcaaa
     1741 aggcacttct acaatggctg ccatcattat tatccgatgt gacgctgcag cttctcaatg
     1801 atattcgaat acgctttgag gagatacagc ctaatatccg acaaactgtt ttacagattt
     1861 acgatcgtac ttgttaccca tcattgaatt ttgaacatcc gaacctggga gttttccctg
     1921 aaacagatag tatatttgaa cctgtataat aatatatagt ctagcgcttt acggaagaca
     1981 atgtatgtat ttcggttcct ggagaaacta ttgcatctat tgcataggta atcttgcacg
     2041 tcgcatcccc ggttcatttt ctgcgtttcc atcttgcact tcaatagcat atctttgtta
     2101 acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc
     2161 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt
     2221 accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga cgagagcgct
     2281 aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc
     2341 gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga
     2401 gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa
     2461 tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt
     2521 ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac
     2581 tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg
     2641 atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc
     2701 agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta
     2761 cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta
     2821 aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa
     2881 ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga
     2941 gatacttttg agcaatgttt gtggaagcgg tattcgcaat gggaagctcc accccggttg
     3001 ataatcagaa aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaattc
     3061 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3121 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3181 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3241 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3301 tatatgagta aacttggtct gacagtcaga agaactcgtc aagaaggcga tagaaggcga
     3361 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     3421 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     3481 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     3541 gcaagcaggc atcgtcatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
     3601 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     3661 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     3721 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     3781 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     3841 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     3901 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     3961 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     4021 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     4081 cggccggaga acctgcgtgc aatccatctt gttcaatcat actcttcctt tttcaatatt
     4141 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4201 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcggacggat
     4261 cgcttgcctg taacttacac gcgcctcgtg gcgcgcccct gcaggattta aatggccggc
     4321 cgtttaaacc ttgtacacct accggtcacc agccaacaat gtgcggatgg cgttacaact
     4381 acaatatgga agggaacttt acacttctcc tatgcacata tattaattaa agtccaatgc
     4441 tagtagagaa ggggggtaac acccctccgc gctcttttcc gatttttttc taaaccgtgg
     4501 aatatttcgg atatcctttt gttgtttccg ggtgtacaat atggacttcc tcttttctgg
     4561 caaccaaacc catacatcgg gattcctata ataccttcgt tggtctccct aacatgtagg
     4621 tggcggaggg gagatataca atagaacaga taccagacaa gacataatgg gctaaacaag
     4681 actacaccaa ttacactgcc tcattgatgg tggtacataa cgaactaata ctgtagccct
     4741 agacttgata gccatcatca tatcgaagtt tcactaccct ttttccattt gccatctatt
     4801 gaagtaataa taggcgcatg caacttcttt tctttttttt tcttttctct ctcccccgtt
     4861 gttgtctcac catatccgca atgacaaaaa aatgatggaa gacactaaag gaaaaaatta
     4921 acgacaaaga cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatctcgaag
     4981 cacacgaaac tttttccttc cttcattcac gcacactact ctctaatgag caacggtata
     5041 cggccttcct tccagttact tgaatttgaa ataaaaaaaa gtttgctgtc ttgctatcaa
     5101 gtataaatag acctgcaatt attaatcttt tgtttcctcg tcattgttct cgttcccttt
     5161 cttccttgtt tctttttctg cacaatattt caagctatac caagcataca atcaactatc
     5221 tcagcggccg catgtctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg
     5281 aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg
     5341 ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg ccagttccat
     5401 ggccaacctt agtcactact ttcggttatg gtgttcaatg ttttgctaga tacccagatc
     5461 atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa
     5521 ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg
     5581 ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt
     5641 taggtcacaa attggaatac aactataact ctcacaatgt ttacatcatg gctgacaaac
     5701 aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggttctgttc
     5761 aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag
     5821 acaaccatta cttatccact caatctgcct tatccaaaga tccaaacgaa aagagagacc
     5881 acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg gatgaattgt
     5941 acaaataagc ggccgcgcga atttcttatg atttatgatt tttattatta aataagttat
     6001 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt
     6061 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc atgaggtcgc
     6121 tcttattgac cacacctagc ttttcaattc aattcatcat ttttttttta ttcttttttt
     6181 tgatttcggt ttctttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     6241 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     6301 aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa
     6361 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgtttgagat
     6421 ttaaatggcg cgccgtttaa accctgcagg ggccggcccc gtagaaaaga tcaaaggatc
     6481 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6541 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6601 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6661 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
//
