LOCUS       40924_22656        9654 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PGREG expression vector pGREG505-G, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9654)
  AUTHORS   Karreman RJ, Morrissey JP.
  TITLE     Controlling expression of genes in Saccharomyces cerevisiae using 
            variants of the pGREG recombinational vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9654)
  AUTHORS   Karreman RJ, Morrissey JP.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JUN-2010) Microbiology Department, University College 
            Cork, College Rd, Cork City, Cork 0000, Ireland
REFERENCE   3  (bases 1 to 9654)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9654)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-JUN-2010) Microbiology Department, University College Cork, 
            College Rd, Cork City, Cork 0000, Ireland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..462
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        762..1166
                     /label=LEU2 promoter
     CDS             1179..2270
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     misc_feature    complement(3005..3508)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        3545..3649
                     /label=AmpR promoter
     CDS             3650..4507
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4681..5269
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5557..5578
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5593..5623
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5631..5647
                     /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     5655..5671
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5692..5710
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      5729..6408
                     /note="Saccharomyces cerevisiae GPD1 promoter (intermediate
                     strength)"
                     /regulatory_class="promoter"
     misc_feature    6433..6466
                     /note="Rec1 site; pGREG recombinational cloning site I"
     misc_feature    6460..6465
                     /label=SalI restriction enzyme cut site
                     /note="SalI restriction enzyme cut site"
     promoter        6755..6942
                     /label=HIS3 promoter
     CDS             6943..7599
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
     misc_feature    7628..7633
                     /label=SalI restriction enzyme cut site
                     /note="SalI restriction enzyme cut site"
     terminator      7640..7887
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    7900..7933
                     /note="LoxP site (cre/lox KanMX marker rescue); Guldener,
                     et. al. 1996, Nucleic Acids Research 24, 2519-2524"
     protein_bind    7900..7933
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            complement(7991..9347)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     protein_bind    complement(9402..9435)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(9472..9490)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9500..9516)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
       61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      421 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcctgatgc ggtattttct
      481 ccttacgcat ctgtgcggta tttcacaccg catatcgacg gtcgaggaga acttctagta
      541 tatccacata cctaatatta ttgccttatt aaaaatggaa tcccaacaat tacatcaaaa
      601 tccacattct cttcaaaatc aattgtcctg tacttccttg ttcatgtgtg ttcaaaaacg
      661 ttatatttat aggataatta tactctattt ctcaacaagt aattggttgt ttggccgagc
      721 ggtctaaggc gcctgattca agaaatatct tgaccgcagt taactgtggg aatactcagg
      781 tatcgtaaga tgcaagagtt cgaatctctt agcaaccatt atttttttcc tcaacataac
      841 gagaacacac aggggcgcta tcgcacagaa tcaaattcga tgactggaaa ttttttgtta
      901 atttcagagg tcgcctgacg catatacctt tttcaactga aaaattggga gaaaaaggaa
      961 aggtgagagg ccggaaccgg cttttcatat agaatagaga agcgttcatg actaaatgct
     1021 tgcatcacaa tacttgaagt tgacaatatt atttaaggac ctattgtttt ttccaatagg
     1081 tggttagcaa tcgtcttact ttctaacttt tcttaccttt tacatttcag caatatatat
     1141 atatatttca aggatatacc attctaatgt ctgcccctat gtctgcccct aagaagatcg
     1201 tcgttttgcc aggtgaccac gttggtcaag aaatcacagc cgaagccatt aaggttctta
     1261 aagctatttc tgatgttcgt tccaatgtca agttcgattt cgaaaatcat ttaattggtg
     1321 gtgctgctat cgatgctaca ggtgtcccac ttccagatga ggcgctggaa gcctccaaga
     1381 aggttgatgc cgttttgtta ggtgctgtgg gtggtcctaa atggggtacc ggtagtgtta
     1441 gacctgaaca aggtttacta aaaatccgta aagaacttca attgtacgcc aacttaagac
     1501 catgtaactt tgcatccgac tctcttttag acttatctcc aatcaagcca caatttgcta
     1561 aaggtactga cttcgttgtt gtcagagaat tagtgggagg tatttacttt ggtaagagaa
     1621 aggaagacga tggtgatggt gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc
     1681 aaagaatcac aagaatggcc gctttcatgg ccctacaaca tgagccacca ttgcctattt
     1741 ggtccttgga taaagctaat gttttggcct cttcaagatt atggagaaaa actgtggagg
     1801 aaaccatcaa gaacgaattc cctacattga aggttcaaca tcaattgatt gattctgccg
     1861 ccatgatcct agttaagaac ccaacccacc taaatggtat tataatcacc agcaacatgt
     1921 ttggtgatat catctccgat gaagcctccg ttatcccagg ttccttgggt ttgttgccat
     1981 ctgcgtcctt ggcctctttg ccagacaaga acaccgcatt tggtttgtac gaaccatgcc
     2041 acggttctgc tccagatttg ccaaagaata aggttgaccc tatcgccact atcttgtctg
     2101 ctgcaatgat gttgaaattg tcattgaact tgcctgaaga aggtaaggcc attgaagatg
     2161 cagttaaaaa ggttttggat gcaggtatca gaactggtga tttaggtggt tccaacagta
     2221 ccaccgaagt cggtgatgct gtcgccgaag aagttaagaa aatccttgct taaaaagatt
     2281 ctcttttttt atgatatttg tacataaact ttataaatga aattcataat agaaacgaca
     2341 cgaaattaca aaatggaata tgttcatagg gtagacgaaa ctatatacgc aatctacata
     2401 catttatcaa gaaggagaaa aaggaggata gtaaaggaat acaggtaagc aaattgatac
     2461 taatggctca acgtgataag gaaaaagaat tgcactttaa cattaatatt gacaaggagg
     2521 agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg aaactacgat tcctaatttg
     2581 atattggagg attttctcta aaaaaaaaaa aatacaacaa ataaaaaaca ctcaatgacc
     2641 tgaccatttg atggagttta agtcaatacc ttcttgaacc atttcccata atggtgaaag
     2701 ttccctcaag aattttactc tgtcagaaac ggccttacga cgtagtcgat atggtgcact
     2761 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     2821 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2881 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     2941 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     3001 gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg
     3061 ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata
     3121 aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc
     3181 aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat
     3241 acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac
     3301 acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg
     3361 tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt
     3421 ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta
     3481 aattataatt atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa
     3541 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3601 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3661 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3721 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3781 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3841 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3901 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3961 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4021 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4081 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4141 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4201 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4261 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4321 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4381 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4441 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4501 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4561 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4621 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4681 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4741 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4801 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4861 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4921 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4981 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5041 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5101 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5161 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5221 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5281 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5341 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5401 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     5461 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     5521 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttacc tcactcatta
     5581 ggcaccccag gctttacact ttatgcttcc ggctcctatg ttgtgtggaa ttgtgagcgg
     5641 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     5701 tcactaaagg gaacaaaagc tggagctctg gggtttgagc aagtctaagt ttacgtagca
     5761 taaaaattct cggattgcgt caaataataa aaaaagtaac cccacttcta cttctacatc
     5821 ggaaaaacat tccattcaca tatcgtcttt ggcctatctt gttttgtcct cggtagatca
     5881 ggtcagtaca aacgcaacac gaaagaacaa aaaaagaaga aaacagaagg ccaagacagg
     5941 gtcaatgaga ctgttgtcct cctactgtcc ctatgtctct ggccgatcac gcgccattgt
     6001 ccctcagaaa caaatcaaac acccacaccc cgggcaccca aagtccccac ccacaccacc
     6061 aatacgtaaa cggggcgccc cctgcaggcc ctcctgcgcg cggcctcccg ccttgcttct
     6121 ctccccttcc ttttcttttt ccagttttcc ctattttgtc cctttttccg cacaacaagt
     6181 atcagaatgg gttcatcaaa tctatccaac ctaattcgca cgtagactgg cttggtattg
     6241 gcagtttcgt agttatatat atactaccat gagtgaaact gttacgttac cttaaattct
     6301 ttctcccttt aattttcttt tatcttactc tcctacataa gacatcaaga aacaattgta
     6361 tattgtacac cccccccctc cacaaacaca aatattgata atataaagac tagtggatcc
     6421 cccgggctgc aggaattcga tatcaagctt atcgataccg tcgacgtcat atggatccgc
     6481 tgcacggtcc tgttccctag catgtacgtg agcgtatttc cttttaaacc acgacgcttt
     6541 gtcttcattc aacgtttccc attgtttttt tctactattg ctttgctgtg ggaaaaactt
     6601 atcgaaagat gacgactttt tcttaattct cgttttaaga gcttggtgag cgctaggagt
     6661 cactgccagg tatcgtttga acacggcatt agtcagggaa gtcataacac agtcctttcc
     6721 cgcaattttc tttttctatt actcttggcc tcctctagta cactctatat ttttttatgc
     6781 ctcggtaatg attttcattt ttttttttcc acctagcgga tgactctttt tttttcttag
     6841 cgattggcat tatcacataa tgaattatac attatataaa gtaatgtgat ttcttcgaag
     6901 aatatactaa aaaatgagca ggcaagataa acgaaggcaa agatgacaga gcagaaagcc
     6961 ctagtaaagc gtattacaaa tgaaaccaag attcagattg cgatctcttt aaagggtggt
     7021 cccctagcga tagagcactc gatcttccca gaaaaagagg cagaagcagt agcagaacag
     7081 gccacacaat cgcaagtgat taacgtccac acaggtatag ggtttctgga ccatatgata
     7141 catgctctgg ccaagcattc cggctggtcg ctaatcgttg agtgcattgg tgacttacac
     7201 atagacgacc atcacaccac tgaagactgc gggattgctc tcggtcaagc ttttaaagag
     7261 gccctactgg cgcgtggagt aaaaaggttt ggatcaggat ttgcgccttt ggatgaggca
     7321 ctttccagag cggtggtaga tctttcgaac aggccgtacg cagttgtcga acttggtttg
     7381 caaagggaga aagtaggaga tctctcttgc gagatgatcc cgcattttct tgaaagcttt
     7441 gcagaggcta gcagaattac cctccacgtt gattgtctgc gaggcaagaa tgatcatcac
     7501 cgtagtgaga gtgcgttcaa ggctcttgcg gttgccataa gagaagccac ctcgcccaat
     7561 ggtaccaacg atgttccctc caccaaaggt gttcttatgt agtgacaccg attatttaaa
     7621 gctgcaggtc gacctcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     7681 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     7741 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     7801 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     7861 ttgggacgct cgaaggcttt aatttgcggc cggtacctaa taacttcgta tagcatacat
     7921 tatacgaagt tatattaagg gttctcgacg ttttcgacac tggatggcgg cgttagtatc
     7981 gaatcgacag cagtatagcg accagcattc acatacgatt gacgcatgat attactttct
     8041 gcgcacttaa cttcgcatct gggcagatga tgtcgaggcg aaaaaaaata taaatcacgc
     8101 taacatttga ttaaaataga acaactacaa tataaaaaaa ctatacaaat gacaagttct
     8161 tgaaaacaag aatcttttta ttgtcagtac tgattagaaa aactcatcga gcatcaaatg
     8221 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     8281 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     8341 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     8401 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
     8461 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     8521 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     8581 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     8641 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     8701 gccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     8761 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     8821 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     8881 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     8941 taaatcagca tccatgttgg aatttaatcg cggcctcgaa acgtgagtct tttccttacc
     9001 catggttgtt tatgttcgga tgtgatgtga gaactgtatc ctagcaagat tttaaaagga
     9061 agtatatgaa agaagaacct cagtggcaaa tcctaacctt ttatatttct ctacaggggc
     9121 gcggcgtggg gacaattcaa cgcgtctgtg aggggagcgt ttccctgctc gcaggtctgc
     9181 agcgaggagc cgtaattttt gcttcgcgcc gtgcggccat caaaatgtat ggatgcaaat
     9241 gattatacat ggggatgtat gggctaaatg tacgggcgac agtcacatca tgcccctgag
     9301 ctgcgcacgt caagactgtc aaggagggta ttctgggcct ccatgtcgct ggccgggtga
     9361 cccggcgggg acgaggcaag ctaaacagat ctctagacct aataacttcg tatagcatac
     9421 attatacgaa gttatattaa gggttgtctt aattaagggt acccaattcg ccctatagtg
     9481 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     9541 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     9601 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaat
//
