LOCUS       40924_22661        9376 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PGREG expression vector pGREG505-T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9376)
  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  JOURNAL   Unpublished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 9376)
  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 9376)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9376)
  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            "Unpublished"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..9376
                     /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"
     promoter        5729..6117
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     misc_feature    6155..6188
                     /note="Rec1 site; pGREG recombinational cloning site"
     misc_feature    6182..6187
                     /label=SalI restriction enzyme cut site
                     /note="SalI restriction enzyme cut site"
     promoter        6477..6664
                     /label=HIS3 promoter
     CDS             6665..7321
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
     misc_feature    7350..7355
                     /label=SalI restriction enzyme cut site
                     /note="SalI restriction enzyme cut site"
     terminator      7362..7609
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     misc_feature    7622..7655
                     /note="LoxP site (cre/lox KanMX marker rescue); Guldener,
                     et. al. 1996, Nucleic Acids Research 24, 2519-2524"
     protein_bind    7622..7655
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            complement(7713..9069)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     protein_bind    complement(9124..9157)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(9194..9212)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9222..9238)
                     /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 tggagctcaa tgtttctact ccttttttac tcttccagat
     5761 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
     5821 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
     5881 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
     5941 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
     6001 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
     6061 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
     6121 taattacaaa actagtggat cccccgggct gcaggaattc gatatcaagc ttatcgatac
     6181 cgtcgacgtc atatggatcc gctgcacggt cctgttccct agcatgtacg tgagcgtatt
     6241 tccttttaaa ccacgacgct ttgtcttcat tcaacgtttc ccattgtttt tttctactat
     6301 tgctttgctg tgggaaaaac ttatcgaaag atgacgactt tttcttaatt ctcgttttaa
     6361 gagcttggtg agcgctagga gtcactgcca ggtatcgttt gaacacggca ttagtcaggg
     6421 aagtcataac acagtccttt cccgcaattt tctttttcta ttactcttgg cctcctctag
     6481 tacactctat atttttttat gcctcggtaa tgattttcat tttttttttt ccacctagcg
     6541 gatgactctt tttttttctt agcgattggc attatcacat aatgaattat acattatata
     6601 aagtaatgtg atttcttcga agaatatact aaaaaatgag caggcaagat aaacgaaggc
     6661 aaagatgaca gagcagaaag ccctagtaaa gcgtattaca aatgaaacca agattcagat
     6721 tgcgatctct ttaaagggtg gtcccctagc gatagagcac tcgatcttcc cagaaaaaga
     6781 ggcagaagca gtagcagaac aggccacaca atcgcaagtg attaacgtcc acacaggtat
     6841 agggtttctg gaccatatga tacatgctct ggccaagcat tccggctggt cgctaatcgt
     6901 tgagtgcatt ggtgacttac acatagacga ccatcacacc actgaagact gcgggattgc
     6961 tctcggtcaa gcttttaaag aggccctact ggcgcgtgga gtaaaaaggt ttggatcagg
     7021 atttgcgcct ttggatgagg cactttccag agcggtggta gatctttcga acaggccgta
     7081 cgcagttgtc gaacttggtt tgcaaaggga gaaagtagga gatctctctt gcgagatgat
     7141 cccgcatttt cttgaaagct ttgcagaggc tagcagaatt accctccacg ttgattgtct
     7201 gcgaggcaag aatgatcatc accgtagtga gagtgcgttc aaggctcttg cggttgccat
     7261 aagagaagcc acctcgccca atggtaccaa cgatgttccc tccaccaaag gtgttcttat
     7321 gtagtgacac cgattattta aagctgcagg tcgacctcga gtcatgtaat tagttatgtc
     7381 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     7441 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     7501 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     7561 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtacct
     7621 aataacttcg tatagcatac attatacgaa gttatattaa gggttctcga cgttttcgac
     7681 actggatggc ggcgttagta tcgaatcgac agcagtatag cgaccagcat tcacatacga
     7741 ttgacgcatg atattacttt ctgcgcactt aacttcgcat ctgggcagat gatgtcgagg
     7801 cgaaaaaaaa tataaatcac gctaacattt gattaaaata gaacaactac aatataaaaa
     7861 aactatacaa atgacaagtt cttgaaaaca agaatctttt tattgtcagt actgattaga
     7921 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     7981 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     8041 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     8101 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     8161 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     8221 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     8281 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     8341 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     8401 ctaatacctg gaatgctgtt ttgccgggga tcgcagtggt gagtaaccat gcatcatcag
     8461 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     8521 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     8581 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     8641 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     8701 aaacgtgagt cttttcctta cccatggttg tttatgttcg gatgtgatgt gagaactgta
     8761 tcctagcaag attttaaaag gaagtatatg aaagaagaac ctcagtggca aatcctaacc
     8821 ttttatattt ctctacaggg gcgcggcgtg gggacaattc aacgcgtctg tgaggggagc
     8881 gtttccctgc tcgcaggtct gcagcgagga gccgtaattt ttgcttcgcg ccgtgcggcc
     8941 atcaaaatgt atggatgcaa atgattatac atggggatgt atgggctaaa tgtacgggcg
     9001 acagtcacat catgcccctg agctgcgcac gtcaagactg tcaaggaggg tattctgggc
     9061 ctccatgtcg ctggccgggt gacccggcgg ggacgaggca agctaaacag atctctagac
     9121 ctaataactt cgtatagcat acattatacg aagttatatt aagggttgtc ttaattaagg
     9181 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     9241 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     9301 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     9361 agcctgaatg gcgaat
//
