LOCUS       40924_34251        6336 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPC_019_GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6336)
  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 6336)
  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 6336)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6336)
  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..6336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(476..1276)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1277..1497)
                     /label=URA3 promoter
     rep_origin      complement(2096..2976)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3325..4116)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4117..4221)
                     /label=AmpR promoter
     promoter        4433..4829
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             4844..5557
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5569..5734
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        5755..5975
                     /label=URA3 promoter
     rep_origin      6096..6336
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
       61 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      121 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
      181 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      241 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      301 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      361 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcatgatat aattaaattg
      421 aagctctaat ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt
      481 gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta
      541 ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg
      601 tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta
      661 aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc
      721 tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct
      781 tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc
      841 ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc
      901 ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag
      961 agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa
     1021 aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag
     1081 tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact
     1141 ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt
     1201 gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat
     1261 atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg
     1321 ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc
     1381 taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat
     1441 ttcaaagaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aattgaaaag
     1501 ctagcttatc gatgataagc tgtcaaagat gagaattaat tccacggact atagactata
     1561 ctagatactc cgtctactgt acgatacact tccgctcagg tccttgtcct ttaacgaggc
     1621 cttaccactc ttttgttact ctattgatcc agctcagcaa aggcagtgtg atctaagatt
     1681 ctatcttcgc gatgtagtaa aactagctag accgagaaag agactagaaa tgcaaaaggc
     1741 acttctacaa tggctgccat cattattatc cgatgtgacg ctgcagcttc tcaatgatat
     1801 tcgaatacgc tttgaggaga tacagcctaa tatccgacaa actgttttac agatttacga
     1861 tcgtacttgt tacccatcat tgaattttga acatccgaac ctgggagttt tccctgaaac
     1921 agatagtata tttgaacctg tataataata tatagtctag cgctttacgg aagacaatgt
     1981 atgtatttcg gttcctggag aaactattgc atctattgca taggtaatct tgcacgtcgc
     2041 atccccggtt cattttctgc gtttccatct tgcacttcaa tagcatatct ttgttaacga
     2101 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     2161 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     2221 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgacgag agcgctaatt
     2281 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
     2341 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
     2401 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
     2461 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
     2521 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
     2581 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
     2641 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
     2701 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
     2761 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
     2821 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
     2881 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
     2941 cttttgagca atgtttgtgg aagcggtatt cgcaatggga agctccaccc cggttgataa
     3001 tcagaaaagc cccaaaaaca ggaagattgt ataagcaaat atttaaattg taattctttc
     3061 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3121 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagggtc
     3181 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3241 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3301 tgagtaaact tggtctgaca gtcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     3361 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     3421 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     3481 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     3541 gcaggcatcg tcatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct
     3601 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     3661 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     3721 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     3781 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     3841 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     3901 cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag
     3961 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     4021 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     4081 cggagaacct gcgtgcaatc catcttgttc aatcatactc ttcctttttc aatattattg
     4141 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4201 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcgg acggatcgct
     4261 tgcctgtaac ttacacgcgc ctcgtggcgc gcccctgcag gatttaaatg gccggccgtt
     4321 taaaccttgt acacctaccg gtcaccagcc aacaatgtgc ggatggcgtt acaactgaag
     4381 tttcactacc ctttttccat ttgccatcta ttgaagtaat aataggcgca tgcaacttct
     4441 tttctttttt tttcttttct ctctcccccg ttgttgtctc accatatccg caatgacaaa
     4501 aaaatgatgg aagacactaa aggaaaaaat taacgacaaa gacagcacca acagatgtcg
     4561 ttgttccaga gctgatgagg ggtatctcga agcacacgaa actttttcct tccttcattc
     4621 acgcacacta ctctctaatg agcaacggta tacggccttc cttccagtta cttgaatttg
     4681 aaataaaaaa aagtttgctg tcttgctatc aagtataaat agacctgcaa ttattaatct
     4741 tttgtttcct cgtcattgtt ctcgttccct ttcttccttg tttctttttc tgcacaatat
     4801 ttcaagctat accaagcata caatcaacta tctcagcggc cgcatgtcta aaggtgaaga
     4861 attattcact ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa
     4921 attttctgtc tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt
     4981 tatttgtact actggtaaat tgccagttcc atggccaacc ttagtcacta ctttcggtta
     5041 tggtgttcaa tgttttgcta gatacccaga tcatatgaaa caacatgact ttttcaagtc
     5101 tgccatgcca gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta
     5161 caagaccaga gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa
     5221 aggtattgat tttaaagaag atggtaacat tttaggtcac aaattggaat acaactataa
     5281 ctctcacaat gtttacatca tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa
     5341 aattagacac aacattgaag atggttctgt tcaattagct gaccattatc aacaaaatac
     5401 tccaattggt gatggtccag tcttgttacc agacaaccat tacttatcca ctcaatctgc
     5461 cttatccaaa gatccaaacg aaaagagaga ccacatggtc ttgttagaat ttgttactgc
     5521 tgctggtatt acccatggta tggatgaatt gtacaaataa gcggccgcgc gaatttctta
     5581 tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa
     5641 agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc
     5701 aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacaccta gcttttcaat
     5761 tcaattcatc attttttttt tattcttttt tttgatttcg gtttctttga aatttttttg
     5821 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     5881 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     5941 acagaacaaa aacctgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     6001 gtgctgctac tcatcctagt cctgtttgag atttaaatgg cgcgccgttt aaaccctgca
     6061 ggggccggcc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6121 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6181 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6241 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6301 tacctcgctc tgctaatcct gttaccagtg gctgct
//
