LOCUS       40924_48308        9956 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pYZ3N-GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9956)
  AUTHORS   Zhao Y, Elder RT, Chen M.
  TITLE     Fission yeast expression vector with GFP tag and positive selection 
            for insert
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9956)
  AUTHORS   Zhao Y, Chen M, Elder RT, Cao J.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-FEB-1998) Departments of Pediatrics and 
            Microbiology-Immunology, Northwestern University Medical School, 
            2430 N Halsted Street, #218, Chicago, IL 60614, USA
REFERENCE   3  (bases 1 to 9956)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9956)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-FEB-1998) Departments of Pediatrics and Microbiology-Immunology,
            Northwestern University Medical School, 2430 N Halsted Street, #218,
            Chicago, IL 60614, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..1210
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces
                     pombe, conferring strong thiamine-repressible expression"
     CDS             1225..1938
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
     primer_bind     2094..2110
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2111..2167
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2180..2196)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2204..2220
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(2228..2258)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2273..2294)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     regulatory      2394..3403
                     /label=from Schizosaccharomyces pombe nmt1
                     /note="from Schizosaccharomyces pombe nmt1"
                     /regulatory_class="terminator"
     rep_origin      complement(3477..4260)
                     /direction=LEFT
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(4610..4626)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4839..5294
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5576..5680
                     /label=AmpR promoter
     CDS             5681..6538
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6712..7300
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7588..7609
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7624..7654
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7662..7678
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     7686..7702
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(8206..9297)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(9310..9714)
                     /label=LEU2 promoter
ORIGIN
        1 agcttgcatg cctgcaggtc gatcgactct agaggatcag aaaattatcg ccataaaaga
       61 cagaataagt catcagcggt tgtttcattt cctatatttt ttttttattt ttttattttt
      121 taataaggga aaatttaacg tctaaggata cagaagattg ttagcacatt aaagtaataa
      181 aggcttaagt agtaagtgcc ttagcatgtt attgtatttc aaaggacata atctaaaata
      241 ataacaatat catttctcac aagttattca attttctttt ttttttctaa taatatcaag
      301 aatgtattat ttgtttgaca taagtcaact aatttattta atatgctgga ttaatcttgc
      361 agacatgtaa attaacaagt tttagtcaaa taacgttgaa gtttcaatga actcaaataa
      421 tttctctttt tttttatata acnnnnnntc taatctgatt tatattttcc gcagggatca
      481 actgaagtta tgacatttgg attggatcac ttataacctt ggtcgccaaa taatacaaaa
      541 atcagcgtta taaaacaaag aaggtttttg ttaagaaatt aatcctcttt cttgataaga
      601 aagttgaacc gaaattgcag atactgatat atgaaaataa tacccacaat tttgggaata
      661 gcgcaagcct caatttaaac aataggtgag gacacatgat aatgacctca atgattgtta
      721 gaagaaaaga gcctcattac aaaatcgaaa aatgaatggt tgggtacaag tttccaaaac
      781 atggtaaagt ggactttgcg tatgagacgt aaatagaaaa aaacacttgt tatatgtttt
      841 ctagaattat tgttgtctct ttatggttgg atgatgcaaa atagtaattt cggttagttg
      901 ctgtaaaaca ccacgagaca aatagatatg gatatttatt aaatcaggaa aaacgtaact
      961 ctcggctact ggatggttca gtcacccaac gattactggg gagagaaaac agggcaaaag
     1021 caaagcttaa aggaatccga ttgtcattcg gcaatgtgca gcgaaactaa aaaccggata
     1081 atggacctgt taatcgaaac attgaagata tataaaggaa gaggaatcct ggcatatcat
     1141 caattgaata agttgaatta attatttcaa tctcattctc actttctgac ttatagtcgc
     1201 tttgttaaat ggcctcgagc caccatgagt aaaggagaag aacttttcac tggagttgtc
     1261 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
     1321 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
     1381 ctacctgttc catggccaac acttgtcact actttcgcgt atggtcttca atgctttgcg
     1441 agatacccag atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
     1501 gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc
     1561 aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa
     1621 gatggaaaca ttcttggaca caaattggaa tacaactata actcacacaa tgtatacatc
     1681 atggcagaca aacaaaagaa tggaatcaaa gttaacttca aaattagaca caacattgaa
     1741 gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     1801 gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac
     1861 gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc
     1921 atggatgaac tatacaaagg taccgcggcc gcgccattcg ccattcaggc tgcgcaactg
     1981 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2041 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2101 gacggccagt gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg
     2161 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2221 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2281 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2341 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgagatcc
     2401 ccgggtaaaa ggaatgtctc ccttgccagt actgctaggg tttttctttc aaactatgga
     2461 agcccattca agctgcatat tacgattttg tttttcgctt ttagaaagtg gtttagatga
     2521 gataatagaa aaattcttga tctccgacaa cgagtacttt tatttttttt gctaatcact
     2581 ttactcaata ttagctcgaa atcgtagaaa cgtagacggg tgcgggatac cgagtggtgt
     2641 agttaagaat ttttataaac cacgtggccc aaaaatatga acccaaaacg tttatacatg
     2701 agtatacttt aagaaggcta taccccttcg tgttagatgt agttttagct acccaacccg
     2761 agtctatgag cttgacttca gatgtagaag gcattaaatc gttttgaata ttaattaaaa
     2821 aacgatgaaa attaaatatt taaaagcaat catacgctga aaatttagtg ctgtggctaa
     2881 tccttcaaca tggaaatgcc ataaaagtga ctttgacaaa aaaaaaagta tatacaggta
     2941 gtaaactcat ctacttcatt gactttgttt acagcatgtg gaaggaggaa tatttattgc
     3001 taaatcgtag tttaacattc aataagtaat actattgaaa ttcgacaaga ttggccgcat
     3061 ggatgaaaaa gaggcatttt gctttgggag aattagttca aattagaact gaaaaaaaaa
     3121 actttacgag gcaaaaatgt cggattgaga tcgtaaaagt tcgctcgtcg tcttttgctt
     3181 tgtgattgtt ttcatggata catcttgctg gatatttaaa ttttagtact atgtataaga
     3241 tattctataa atgttttatc acccaaacct gttagcgcct tcttaattct attcaatctg
     3301 gcttttgctc tgagactact tcttggactt tcactacttg ttagttatac ggaatttgtg
     3361 taattagaag tgaaataatc ctttctatta gtaatgcgag ctcgaattcg agtctaactc
     3421 cttaaccact cggacatagt gacttatctg acactcattg taaattaata tatatatagg
     3481 cattttgttt agttaaaggt acttaagtaa ttagtataaa cgaaccaatt ttataatcag
     3541 gaagttaagt gaatggtagc acatgtcgta aaaattgtga atttttattg aataatattt
     3601 taaatacaag cctttctaga ctaggtatac tcataaacat atatgagcaa aaggatagag
     3661 gagattacat tgcatcttct acaaatttat ttattgccct ttactgaaaa attaaataat
     3721 gagtactaaa tgataaaaag cgctcagtac aagaaagatt gcaaaaatat tgcattcttc
     3781 atgaattaaa gttgcatata aggcatattg aaagtaatag tactaaaaca gcagttagcg
     3841 aaaattaata gaattatatt cgcaagacaa ttgtgacatt aaattaaaaa attgtagaat
     3901 ttttactatc ctctttaacg ccatgagcct ttataaaaag gttaaattag ttttaacatt
     3961 ctttttttga gtaagagttt acaatttatc aaaacctgtg ttattatatt cattagtttc
     4021 aatttattag catctagaga aaaatcaatt ggcagttaca ttgttggaat ttatgaagaa
     4081 aaagaatcta caacggagaa taagttgctg atcgctttcc ctaaaattgt atattttgct
     4141 gagcttattt tgacatttcg ttgaagtttt ctctaattcg cattcatttt aagtaaaaca
     4201 atgagaaata aaattacaaa aaatacaaat taaaatacaa tttttagcta taattataga
     4261 cgatgccctt gtatcccatt ctgtctcgct tgcccctact ttttatcttt tatataccat
     4321 aatgaacgct gccgctacta accatacccc gattttacat ttcggactcc caaggacgta
     4381 caaaatagaa aactatagaa aaaaataatc agaaaatagc atgtcatctc tttgtaaaac
     4441 gcgtttgcaa gaagaaagga aacaatggag aagagatcat ccatttgtat gtaaatttta
     4501 gtaaacttga agaaatcact aacaacttct cttacttagg gattctatgc aaaaccttgt
     4561 aaatcatctg atggaggact cgatttaatg aattggaagg ttggaattca ctggccgtcg
     4621 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4681 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4741 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     4801 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4861 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4921 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4981 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5041 caaaaaactt gatttgggtg atggttcacg tagtgggaca tcgccctgat agacggtttt
     5101 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5161 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     5221 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5281 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5341 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5401 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5461 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5521 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5581 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5641 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5701 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5761 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5821 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5881 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5941 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6001 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6061 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6121 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6181 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6241 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6301 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6361 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6421 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6481 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6541 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6601 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6661 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6721 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6781 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6841 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6901 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6961 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7021 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7081 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7141 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7201 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7261 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7321 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7381 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7441 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     7501 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     7561 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     7621 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     7681 tcacacagga aacagctatg accatgatta cgccaagctt gtcgatcgac tacgtcgtta
     7741 aggccgtttc tgacagagta aaattcttga gggaactttc accattatgg gaaatggttc
     7801 aagaaggtat tgacttaaac tccatcaaat ggtcaggtca ttgagtgttt tttatttgtt
     7861 gtattttttt ttttttagag aaaatcctcc aatatcaaat taggaatcgt agtttcatga
     7921 ttttctgtta cacctaactt tttgtgtggt gccctcctcc ttgtcaatat taatgttaaa
     7981 gtgcaattct ttttccttat cacgttgagc cattagtatc aatttgctta cctgtattcc
     8041 tttactatcc tcctttttct ccttcttgat aaatgtatgt agattgcgta tatagtttcg
     8101 tctaccctat gaacatattc cattttgtaa tttcgtgtcg tttctattat gaatttcatt
     8161 tataaagttt atgtacaaat atcataaaaa aagagaatct ttttaagcaa ggattttctt
     8221 aacttcttcg gcgacagcat caccgacttc ggtggtactg ttggaaccac ctaaatcacc
     8281 agttctgata cctgcatcca aaaccttttt aactgcatct tcaatggcct taccttcttc
     8341 aggcaagttc aatgacaatt tcaacatcat tgcagcagac aagatagtgg cgatagggtc
     8401 aaccttattc tttggcaaat ctggagcaga accgtggcat ggttcgtaca aaccaaatgc
     8461 ggtgttcttg tctggcaaag aggccaagga cgcagatggc aacaaaccca aggaacctgg
     8521 gataacggag gcttcatcgg agatgatatc accaaacatg ttgctggtga ttataatacc
     8581 atttaggtgg gttgggttct taactaggat catggcggca gaatcaatca attgatgttg
     8641 aaccttcaat gtagggaatt cgttcttgat ggtttcctcc acagtttttc tccataatct
     8701 tgaagaggcc aaaacattag ctttatccaa ggaccaaata ggcaatggtg gctcatgttg
     8761 tagggccatg aaagcggcca ttcttgtgat tctttgcact tctggaacgg tgtattgttc
     8821 actatcccaa gcgacaccat caccatcgtc ttcctttctc ttaccaaagt aaatacctcc
     8881 cactaattct ctgacaacaa cgaagtcagt acctttagca aattgtggct tgattggaga
     8941 taagtctaaa agagagtcgg atgcaaagtt acatggtctt aagttggcgt acaattgaag
     9001 ttctttacgg atttttagta aaccttgttc aggtctaaca ctaccggtac cccatttagg
     9061 accacccaca gcacctaaca aaacggcatc aaccttcttg gaggcttcca gcgcctcatc
     9121 tggaagtggg acacctgtag catcgatagc agcaccacca attaaatgat tttcgaaatc
     9181 gaacttgaca ttggaacgaa catcagaaat agctttaaga accttaatgg cttcggctgt
     9241 gatttcttga ccaacgtggt cacctggcaa aacgacgatc ttcttagggg cagacatagg
     9301 ggcagacatt agaatggtat atccttgaaa tatatatata tattgctgaa atgtaaaagg
     9361 taagaaaagt tagaaagtaa gacgattgct aaccacctat tggaaaaaac aataggtcct
     9421 taaataatat tgtcaacttc aagtattgtg atgcaagcat ttagtcatga acgcttctct
     9481 attctatatg aaaagccggt tccggcctct cacctttcct ttttctccca atttttcagt
     9541 tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa
     9601 tttgattctg tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg
     9661 ttgctaagag attcgaactc ttgcatctta cgatacctga gtattcccac agttaactgc
     9721 ggtcaagata tttcttgaat caggcgcctt agaccgctcg gccaaacaac caattacttg
     9781 ttgagaaata gagtataatt atcctataaa tataacgttt ttgaacacac atgaacaagg
     9841 aagtacagga caattgattt tgaagagaat gtggattttg atgtaattgt tgggattcca
     9901 tttttaataa ggcaataata ttaggtatgt agatatacta gaagttctcc tcgaca
//
