LOCUS       40924_45483        5785 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PCR template vector pUG30, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5785)
  AUTHORS   Gueldener U, Hegemann JH.
  TITLE     A second generation of GFP-vectors for subcelluar localization 
            studies in budding yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5785)
  AUTHORS   Gueldener U, Hegemann JH.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany
REFERENCE   3  (bases 1 to 5785)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5785)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    67..88
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        103..133
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    141..157
                     /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     165..181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        202..220
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        271..620
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     621..637
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             676..1389
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1411..1658
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1677..1695)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1705..1721)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1829..1862
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            1917..3273
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     misc_feature    3336..3369
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(3336..3369)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(3423..3441)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(3699..4287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4461..5318)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5319..5423)
                     /label=AmpR promoter
     promoter        5769..5785
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
       61 cgcaattaat gtgagttacc tcactcatta ggcaccccag gctttacact ttatgcttcc
      121 ggctcctatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      181 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctccg
      241 gatgcaaggg ttcgaatccc ttagctctca ttattttttg ctttttctct tgaggtcaca
      301 tgatcgcaaa atggcaaatg gcacgtgaag ctgtcgatat tggggaactg tggtggttgg
      361 caaatgacta attaagttag tcaaggcgcc atcctcatga aaactgtgta acataataac
      421 cgaagtgtcg aaaaggtggc accttgtcca attgaacacg ctcgatgaaa aaaataagat
      481 atatataagg ttaagtaaag cgtctgttag aaaggaagtt tttccttttt cttgctctct
      541 tgtcttttca tctactattt ccttcgtgta atacagggtc gtcagataca tagatacaat
      601 tctattaccc ccatccatac tctagaacta gtggatcccc cgggctgcag gaattcgata
      661 tcaagcttta ttaaaatgtc taaaggtgaa gaattattca ctggtgttgt cccaattttg
      721 gttgaattag atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt
      781 gatgctactt acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccagtt
      841 ccatggccaa ccttagtcac tactttcggt tatggtgttc aatgttttgc tagataccca
      901 gatcatatga aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa
      961 agaactattt ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa
     1021 ggtgatacct tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac
     1081 attttaggtc acaaattgga atacaactat aactctcaca atgtttacat catggctgac
     1141 aaacaaaaga atggtatcaa agttaacttc aaaattagac acaacattga agatggttct
     1201 gttcaattag ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta
     1261 ccagacaacc attacttatc cactcaatct gccttatcca aagatccaaa cgaaaagaga
     1321 gaccacatgg tcttgttaga atttgttact gctgctggta ttacccatgg tatggatgaa
     1381 ttgtacaaat aactgcaggt cgacctcgag tcatgtaatt agttatgtca cgcttacatt
     1441 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     1501 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     1561 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     1621 ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta
     1681 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1741 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctgaagctt
     1801 cgtacgctgc aggtcgacaa cccttaatat aacttcgtat aatgtatgct atacgaagtt
     1861 attaggtcta gagatctgtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca
     1921 tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg
     1981 actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt
     2041 ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc tgcgagcagg
     2101 gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata
     2161 taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc
     2221 taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg aaaagactca
     2281 cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2341 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
     2401 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2461 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2521 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag cattccaggt
     2581 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2641 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2701 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2761 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc
     2821 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2881 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2941 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3001 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3061 gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     3121 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     3181 tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc
     3241 gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat
     3301 ccagtgtcga aaacgagctc tcgagaaccc ttaatataac ttcgtataat gtatgctata
     3361 cgaagttatt aggtgatatc agatccacta gtggcctatg cggccgcgga tctgccggtc
     3421 tccctatagt gagtcgtatt aatttcgata agccaggtta acctgcatta atgaatcggc
     3481 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3541 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3601 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3661 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3721 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3781 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3841 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     3901 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3961 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4021 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4081 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     4141 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4201 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4261 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4321 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4381 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4441 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4501 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4561 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4621 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4681 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4741 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4801 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4861 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4921 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4981 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5041 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5101 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5161 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5221 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5281 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5341 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5401 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5461 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5521 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5581 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5641 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     5701 catatggaca tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata
     5761 cacatacgat ttaggtgaca ctata
//
