LOCUS       40924_10906        5991 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCK908, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5991)
  AUTHORS   Kraft C.
  TITLE     A toolbox for the analysis of weak and low abundant molecule 
            interactions in vivo by M-Track in budding yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5991)
  AUTHORS   Brezovich A, Schuschnig M, Ammerer G, Kraft C.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2014) Department of Biochemistry and Cell Biology,
            MFPL, University of Vienna, Dr. Bohr Gasse 9, Vienna 1030, Austria
REFERENCE   3  (bases 1 to 5991)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5991)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-AUG-2014) Department of Biochemistry and Cell Biology, MFPL, 
            University of Vienna, Dr. Bohr Gasse 9, Vienna 1030, Austria"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    167..188
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        203..233
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    241..257
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     265..281
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        302..320
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        344..785
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     misc_feature    815..1108
                     /label=4x Histone3
                     /note="4x Histone3"
     CDS             1142..1231
                     /codon_start=1
                     /label=3xHA
                     /note="three tandem HA epitope tags"
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(1261..1277)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1280..1527
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1546..1564)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1574..1590)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1731..2186
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2320..3120)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3121..3336)
                     /label=URA3 promoter
     misc_feature    complement(3606..4109)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        4146..4250
                     /label=AmpR promoter
     CDS             4251..5108
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5282..5870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
       61 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
      121 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttacc
      181 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcctatg ttgtgtggaa
      241 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      301 caattaaccc tcactaaagg gaacaaaagc tggagctcta gtacggatta gaagccgccg
      361 agcgggtgac agccctccga aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg
      421 cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa
      481 tactagcttt tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc
      541 aaatgaacga atcaaattaa caaccatagg atgataatgc gattagtttt ttagccttat
      601 ttctggggta attaatcagc gaagcgatga tttttgatct attaacagat atataaatgc
      661 aaaaactgca taaccacttt aactaatact ttcaacattt tcggtttgta ttacttctta
      721 ttcaaatgta ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca
      781 aggagaaaaa accccggatt ctagaactag tatggccaga accaagcaaa cagctaggaa
      841 atcaactggt ggtaaagcac ctcgtaaaca attagctcag ataccaaata gtgccagaac
      901 caaacaaact gcgaggaaaa gtacaggagg caaagctcct agaaaacaat tagcccaaat
      961 acccaattct gcaagaacaa agcaaacagc gagaaaatct accggtggta aggccccaag
     1021 aaagcagtta gcacaaattc caaactcggc tagaacgaaa caaacggcta gaaagtcaac
     1081 tggaggtaaa gcaccaagga aacagttgac tagctcttta ggggatggat ctcctaggat
     1141 gtacccatac gatgttcctg actatgcggg ctatccctat gacgtcccgg actatgcagg
     1201 ttcctatcca tatgacgttc cagattacgc tggcctgcag gaattcgata tcaagcttat
     1261 cgataccgtc gacctcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     1321 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1381 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1441 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     1501 ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt
     1561 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     1621 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     1681 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     1741 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1801 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1861 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1921 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1981 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2041 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2101 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2161 taacaaaata ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     2221 ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc taatttgtga
     2281 gtttagtata catgcattta cttataatac agttttttag ttttgctggc cgcatcttct
     2341 caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc
     2401 ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc
     2461 cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt
     2521 cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc
     2581 gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt
     2641 agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt
     2701 tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc
     2761 attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac
     2821 tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga
     2881 taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg
     2941 tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt
     3001 ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag
     3061 cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat
     3121 gatttatctt cgtttcctgc atgtttttgt tctgtgcagt tgggttaaga atactgggca
     3181 atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt
     3241 ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaaa gaaaccgaaa
     3301 tcaaaaaaaa gaataaaaaa aaaatgatga attgaattga aaagctgtgg tatggtgcac
     3361 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3421 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3481 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3541 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3601 ggacggatcg cttgcctgta acttacacgc gcctcgtatc ttttaatgat ggaataattt
     3661 gggaatttac tctgtgttta tttattttta tgttttgtat ttggatttta gaaagtaaat
     3721 aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt
     3781 caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat taaatagata
     3841 tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta
     3901 cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc aagataaaag
     3961 gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca aaaactattt
     4021 tttctttaat ttcttttttt actttctatt tttaatttat atatttatat taaaaaattt
     4081 aaattataat tatttttata gcacgtgatg aaaaggaccc aggtggcact tttcggggaa
     4141 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4201 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4261 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4321 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4381 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4441 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4501 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4561 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4621 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4681 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4741 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4801 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4861 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4921 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4981 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5041 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5101 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5161 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5221 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5281 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5341 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5401 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5461 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5521 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5581 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5641 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5701 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5761 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5821 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5881 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5941 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct g
//
