LOCUS       40924_48228        8332 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYOGM081, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8332)
  AUTHORS   Labunskyy VM, Suzuki Y, Hanly TJ, Murao A, Roth FP, Gladyshev VN.
  TITLE     The insertion Green Monster (iGM) method for expression of multiple 
            exogenous genes in yeast
  JOURNAL   G3 (Bethesda) 4 (7), 1183-1191 (2014)
  PUBMED    24776987
REFERENCE   2  (bases 1 to 8332)
  AUTHORS   Suzuki Y, Roth FP.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JAN-2014) Synthetic Biology and Bioenergy, J. Craig 
            Venter Institute, 4120 Torrey Pines Road, La Jolla, CA 92037, USA
REFERENCE   3  (bases 1 to 8332)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8332)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda)"; date: "2014"; volume: "4"; issue: "7"; pages: 
            "1183-1191"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JAN-2014) Synthetic Biology and Bioenergy, J. Craig Venter 
            Institute, 4120 Torrey Pines Road, La Jolla, CA 92037, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    6..70
                     /label=upstream linearization site
                     /note="upstream linearization site"
     misc_feature    33..152
                     /label=similar to the KanMX4 cassette
                     /note="similar to the KanMX4 cassette"
     promoter        167..383
                     /label=URA3 promoter
     CDS             384..1184
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     terminator      1295..1482
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     protein_bind    1499..1517
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1541..1559
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      1594..1746
                     /label=CYC1 TATA region
                     /note="CYC1 TATA region"
                     /regulatory_class="TATA_box"
     CDS             1755..2468
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
     terminator      2493..2740
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        2756..3197
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     3212..3228
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3236..3360
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3385..3415
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3469..4125
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4473..4775
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(4819..4943)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4984..5073
                     /label=3xHA
                     /note="three tandem HA epitope tags"
     terminator      5107..5304
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    5346..5404
                     /label=downstream linearization site
                     /note="downstream linearization site"
     primer_bind     complement(5424..5440)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5477..5495)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5516..5532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5540..5556)
                     /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)."
     promoter        complement(5564..5594)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5609..5630)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5918..6506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6680..7537)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7538..7642)
                     /label=AmpR promoter
     rep_origin      complement(7668..8123)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8265..8281
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8291..8309
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ctcgagctag ccgcggtacg taccaggtga gctcgacgga tccccgggtt aattaaggcg
       61 cgccagatct gtttagcttg ccttgtcccc gccgggtcac ccggccagcg acatggaggc
      121 ccagaatacc ctccttgaca gtcttgacgt gcgtcgagct tttcaattca attcatcttt
      181 tttttttttg ttcttttttt tgattccggt ttctttgaaa tttttttgat tcggtaatct
      241 ccgagcagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
      301 gtggtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
      361 cctgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
      421 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
      481 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
      541 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
      601 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa
      661 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
      721 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
      781 tgaagcaggc ggcggaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
      841 tgtcatgcaa gggctcccta gctactggag aatatactaa gggtactgtt gacattgcga
      901 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
      961 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat
     1021 tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg
     1081 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     1141 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     1201 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     1261 tacccgtcga gtaattcgcg ccacttctaa ataagcgaat ttcttatgat ttatgatttt
     1321 tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt
     1381 tttaaaacga aaattcttgt tcttgagtaa ctctttcctg taggtcaggt tgctttctca
     1441 ggtatagcat gaggtcgctc ttattgacca cacctctacc ggcagatcaa ttcctcgatc
     1501 cctatcagtg atagagagtc gacaaagtcg agtttctcga tccctatcag tgatagagag
     1561 tcgacaaagt cgagtttctc gatcgagacc actgcatgca tgtgctctgt atgtatataa
     1621 aactcttgtt ttcttctttt ctctaaatat tctttcctta tacattaggt cctttgtagc
     1681 ataaattact atacttctat agacacgcaa acacaaatac acacactaaa ttaccggatc
     1741 aattcggggg atccatgagt aaaggagaag aacttttcac tggagtggtc ccagttcttg
     1801 ttgaattaga tggcgatgtt aatgggcaaa aattctctgt cagtggagag ggtgaaggtg
     1861 atgcaacata cggaaaactt acccttaatt ttatttgcac tactgggaag ctacctgttc
     1921 catggccaac acttgtcact actttctctt atggtgttca atgcttctca agatacccag
     1981 atcatatgaa acagcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa
     2041 gaactatatt ttacaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag
     2101 gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca
     2161 ttcttggaca caaaatggaa tacaactata actcacataa tgtatacatc atgggagaca
     2221 aaccaaagaa tggcatcaaa gttaacttca aaattagaca caacattaaa gatggaagcg
     2281 ttcaattagc agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac
     2341 cagacaacca ttacctgtcc acacaatctg ccctttccaa agatcccaac gaaaagagag
     2401 atcacatgat ccttcttgag tttgtaacag ctgctaggat tacacatggc atggatgaac
     2461 tatacaaata agggccctgc aggagggccg catcatgtaa ttagttatgt cacgcttaca
     2521 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     2581 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     2641 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     2701 gcttgagaag gttttgggac gctcgaaggc tttaatttgc ggccaagctc ctgtacggat
     2761 tagaagccgc cgagcgggcg acagccctcc gacggaagac tctcctccgt gcgtcctcgt
     2821 cttcaccggt cgcgttcctg aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata
     2881 aagattctac aatactagct tttatggtta tgaagaggaa aaattggcag taacctggcc
     2941 ccacaaacct tcaaattaac gaatcaaatt aacaaccata ggatgataat gcgattagtt
     3001 ttttagcctt atttctgggg taattaatca gcgaagcgat gatttttgat ctattaacag
     3061 atatataaat ggaaaagctg cataaccact ttaactaata ctttcaacat tttcagtttg
     3121 tattacttct tattcaaatg tcataaaagt atcaacaaaa aattgttaat atacctctat
     3181 actttaacgt caaggagaaa aaaccccgga ttctagaact agtggatccc ccatcacaag
     3241 tttgtacaaa aaagctgaac gagaaacgta aaatgatata aatatcaata tattaaatta
     3301 gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg
     3361 gcggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg
     3421 attttgagtt aggatccgtc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     3481 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     3541 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     3601 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     3661 cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga gctggtgata
     3721 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     3781 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     3841 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     3901 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     3961 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     4021 atgccgctgg cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt cggcagaatg
     4081 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaaac gccgcgtgga
     4141 tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata
     4201 agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc
     4261 gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa
     4321 tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg
     4381 ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg
     4441 ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa
     4501 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc
     4561 gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac
     4621 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca
     4681 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca
     4741 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca
     4801 gccagtctgc aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc
     4861 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt
     4921 tcagctttct tgtacaaagt ggtgatgggc tgcaggaatt cgatatcaag cttaggtgga
     4981 atgtacccat acgatgttcc tgactatgcg ggctatccct atgacgtccc ggactatgca
     5041 ggatcctatc catatgacgt tccagattac gctgctcagt gctaactcga gtcaggagct
     5101 ttcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     5161 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     5221 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     5281 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgtcg
     5341 aaaacgagct cagatctggc gcgccgctag cgagctctac gtattaatta accgcggacc
     5401 tggtgaattc ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggtgga
     5461 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     5521 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5581 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5641 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5701 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5761 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5821 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5881 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5941 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6001 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6061 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6121 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6181 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6241 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6301 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6361 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6421 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6481 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6541 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6601 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6661 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6721 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6781 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6841 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6901 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6961 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7021 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7081 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7141 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7201 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7261 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7321 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7381 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7441 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7501 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7561 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7621 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     7681 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     7741 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     7801 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     7861 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     7921 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     7981 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     8041 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     8101 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
     8161 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     8221 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     8281 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc cc
//
