LOCUS       sequence-198979-        8131 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Pombe Expression Vector - nmt81 promoter with Sp.ura4 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8131)
  AUTHORS   Zhao Y, Boeke JD
  TITLE     Construction of Designer Selectable Marker Deletions with 
            CRISPR-Cas9 Toolbox in Schizosaccharomyces pombe and Optimized 
            Design of Common Entry Vectors.
  JOURNAL   G3 (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 
            10.1534/g3.117.300363.
  PUBMED    29321167
REFERENCE   2  (bases 1 to 8131)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda). 2018 Jan 10. pii: g3.117.300363. doi: 
            10.1534/g3.117.300363."
COMMENT     Sequence Label: pYZ157
FEATURES             Location/Qualifiers
     source          1..8131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             217..597
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISRLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    273..380
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     complement(388..407)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(389..407)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(417..434)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(417..433)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(426..448)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      574..1029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(661..680)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     871..892
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1465..2259)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     promoter        2818..3969
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     rep_origin      5067..5850
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6226..6330
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6331..6399,6400..7191)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6549..6568)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7362..7950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7851..7870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8104..8121
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctcta
      301 gaactagtcg atccgccggg ctgcaggaat tcgatatcaa ggttatcgat accgtccacc
      361 tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
      421 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
      481 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
      541 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
      601 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
      661 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      721 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
      781 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      841 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      901 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
      961 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     1021 ttacaattta gcttgtgata ttgacgaaac tttttgacat ctaatttatt ctgttccaac
     1081 accaatgttt ataaccaagt tttatcttgt ttgtctacat ggtattttac attcatctac
     1141 atacatcttt cattggcttt gtacatagtt atcattacaa gtctaaaaaa attcactctt
     1201 ttcttattca atgtcaatcc aagagaaaag attgtggtaa tgttgtagga gcatgtttaa
     1261 taaattacta tagcaaatta ctttttattc ccaaggtgtt tatctataat agttaatatt
     1321 ttagtcgcta cataaaattt taccaaagag tacttgtata ctaattctaa atgccttctg
     1381 acataaaacg cctaggaaaa caaacgcaaa caaggcatcg actttttcaa taaccaacca
     1441 aaaaaatttt acattagtct ttttttaatg ctgagaaagt ctttgctgat atgccttcca
     1501 accagcttct ctatatctct tggcttcgac aacaggatta cgaccagctc catagactcc
     1561 acgaccaaca atgatgatat cgctaccgca gtttacaatc acttcttcag gagtacgata
     1621 ttgctgtccc agcccgtctc ctttaacatc caagccgata ccaggggaca tagttatgta
     1681 gtcgctttga aggttaggaa atcgacgacc agctataaag ccaaagcaaa aatcggtatg
     1741 cttctcaaac cattctaagg ttttctctgt gtaggaacca gtagccaaag agcctttgga
     1801 agacatttca gccaaaagca agagtccacg tcccaaaggt aaaccaactt ctttgaggcc
     1861 ttgtataata ccctcgcctg gcactgtatg gcaatttgtg atatgagccc aagaagcaat
     1921 tttgtacaca ccagatgcat attgtagctt gacggtattt ccaatgtctg cgaatttgcg
     1981 atcctcaaag ataagaaaac gatgcttttt acctaaggcc accagttttt ctaccatatc
     2041 ctggtcgaaa tcctcgacaa cgtcaatatg tgtcttgata acacagacat agggtccaat
     2101 tttatctacc aattctaaga tttcggattt cttcgtcaaa tcgaccgcga ctgacaagtt
     2161 gctttgcttt tcttccatca aagccaacaa ttccttggca atgggatttt tcatcccctc
     2221 agctctagct gaatagcttt gaaatactct agcatccata actttgcttt taaaccttta
     2281 atttcgatcc aagcaaaaaa gaggttcttg gtaggacaat acggtaagaa aacacgacat
     2341 gtgcagagat gccgacgaag catagttaaa ctgggatggt aaaatcaatt aagaatttat
     2401 aaagacaaaa ttgtataagt ctctaaaaca tcttaattat acctcacaga actatctaaa
     2461 atatattcac aaagtgcaaa cattatcatg aaaaagaacc attttaattt aaagcaaggg
     2521 cattaaggct tatttacaga atttcttact tttgtaaaga ttataaggct gattatcttt
     2581 ttcaccatgc caaaaattac acaagataga atggatgttt gaaattaaac gtgagtatac
     2641 aaacaaatac actaggtaaa tcgaaacatt tttttctcca ttaagtaaca aattcctatt
     2701 tagagaaaga atgctgagta gattaaataa tctatacaaa cttttttaac acaaatgcat
     2761 acatatagcc agtgggattt gtagctagtc gatcgactct agaggatcag aaaattatcg
     2821 ccataaaaga cagaataagt catcagcggt tgtttcattt cctatatttt ttttttattt
     2881 ttttattttt taataaggga aaatttaacg tctaaggata cagaagattg ttagcacatt
     2941 aaagtaataa aggcttaagt agtaagtgcc ttagcatgtt attgtatttc aaaggacata
     3001 atctaaaata ataacaatat catttctcac aagttattca attttctttt ttttttctaa
     3061 taatatcaag aatgtattat ttgtttgaca taagtcaact aatttattta atatgctgga
     3121 ttaatcttgc agacatgtaa attaacaagt tttagtcaaa taacgttgaa gtttcaatga
     3181 actcaaataa tttctctttt tttttatata accataatct gatttatatt ttccgcaggg
     3241 atcaactgaa gttatgacat ttggattgga tcacttataa ccttggtcgc caaataatac
     3301 aaaaatcagc gttataaaac aaagaaggtt tttgttaaga aattaatcct ctttcttgat
     3361 aagaaagttg aaccgaaatt gcagatactg atatatgaaa ataataccca caattttggg
     3421 aatagcgcaa gcctcaattt aaacaatagg tgaggacaca tgataatgac ctcaatgatt
     3481 gttagaagaa aagagcctca ttacaaaatc gaaaaatgaa tggttgggta caagtttcca
     3541 aaacatggta aagtggactt tgcgtatgag acgtaaatag aaaaaaacac ttgttatatg
     3601 ttttctagaa ttattgttgt ctctttatgg ttggatgatg caaaatagta atttcggtta
     3661 gttgctgtaa aacaccacga gacaaataga tatggatatt tattaaatca ggaaaaacgt
     3721 aactctcggc tactggatgg ttcagtcacc caacgattac tggggagaga aaacagggca
     3781 aaagcaaagc ttaaaggaat ccgattgtca ttcggcaatg tgcagcgaaa ctaaaaaccg
     3841 gataatggac ctgttaatcg aaacattgaa gatggaagag gaatcctggc atatcatcaa
     3901 ttgaataagt tgaattaatt atttcaatct cattctcact ttctgactta tagtcgcttt
     3961 gttaaatcat atgtcgactc tagaggatcc ccgggtaaaa ggaatgtctc ccttgccagt
     4021 actgctaggg tttttctttc aaactatgga agcccattca agctgcatat tacgattttg
     4081 tttttcgctt ttagaaagtg gtttagatga gataatagaa aaattcttga tctccgacaa
     4141 cgagtacttt tatttttttt gctaatcact ttactcaata ttagctcgaa atcgtagaaa
     4201 cgtagacggg tgcgggatac cgagtggtgt agttaagaat ttttataaac cacgtggccc
     4261 aaaaatatga acccaaaacg tttatacatg agtatacttt aagaaggcta taccccttcg
     4321 tgttagatgt agttttagct acccaacccg agtctatgag cttgacttca gatgtagaag
     4381 gcattaaatc gttttgaata ttaattaaaa aacgatgaaa attaaatatt taaaagcaat
     4441 catacgctga aaatttagtg ctgtggctaa tccttcaaca tggaaatgcc ataaaagtga
     4501 ctttgacaaa aaaaaaagta tatacaggta gtaaactcat ctacttcatt gactttgttt
     4561 acagcatgtg gaaggaggaa tatttattgc taaatcgtag tttaacattc aataagtaat
     4621 actattgaaa ttcgacaaga ttggccgcat ggatgaaaaa gaggcatttt gctttgggag
     4681 aattagtcca aattagaact gaaaaaaaaa actttacgag gcaaaaatgt cggattgaga
     4741 tcgtaaaagt tcgctcgtcg tcttttgctt tgtgattgtt ttcatggata catcttgctg
     4801 gatatttaaa ttttagtact atgtataaga tattctataa atgttttatc acccaaacct
     4861 gttagcgcct tcttaattct attcaatctg gcttttgctc tgagactact tcttggactt
     4921 tcactacttg ttagttatac ggaatttgtg taattagaag tgaaataatc ctttctatta
     4981 gtaatgcgag ctgcaattcg agtctaactc cttaaccact cggacatagt gacttatctg
     5041 acactcattg taaattaata tatatatagg cattttgttt agttaaaggt acttaagtaa
     5101 ttagtataaa cgaaccaatt ttataatcag gaagttaagt gaatggtagc acatgtcgta
     5161 aaaattgtga atttttattg aataatattt taaatacaag cctttctaga ctaggtatac
     5221 tcataaacat atatgagcaa aaggatagag gagattacat tgcatcttct acaaatttat
     5281 ttattgccct ttactgaaaa attaaataat gagtactaaa tgataaaaag cgctcagtac
     5341 aagaaagatt gcaaaaatat tgcattcttc atgaattaaa gttgcatata aggcatattg
     5401 aaagtaatag tactaaaaca gcagttagcg aaaattaata gaattatatt cgcaagacaa
     5461 ttgtgacatt aaattaaaaa attgtagaat ttttactatc ctctttaacg ccatgagcct
     5521 ttataaaaag gttaaattag ttttaacatt ctttttttga gtaagagttt acaatttatc
     5581 aaaacctgtg ttattatatt cattagtttc aatttattag catctagaga aaaatcaatt
     5641 ggcagttaca ttgttggaat ttatgaagaa aaagaatcta caacggagaa taagttgctg
     5701 atcgctttcc ctaaaattgt atattttgct gagcttattt tgacatttcg ttgaagtttt
     5761 ctctaattcg cattcatttt aagtaaaaca atgagaaata aaattacaaa aaatacaaat
     5821 taaaatacaa tttttagcta taattataga cgatgccctt gtatcccatt ctgtctcgct
     5881 tgcccctact ttttatcttt tatataccat aatgaacgct gccgctacta accatacccc
     5941 gattttacat ttcggactcc caaggacgta caaaatagaa aactatagaa aaaaataatc
     6001 agaaaatagc atgtcatctc tttgtaaaac gcgtttgcaa gaagaaagga aacaatggag
     6061 aagagatcat ccatttgtat gtaaatttta gtaaacttga agaaatcact aacaacttct
     6121 cttacttagg gattctatgc aaaaccttgt aaatcatctg atggaggact cgatttaatg
     6181 aattggaagg ttggatttcc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6241 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6301 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6361 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6421 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6481 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6541 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6601 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6661 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6721 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6781 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6841 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6901 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6961 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7021 gataaatctg gagccggtga gcgtggatct cgcggtatca ttgcagcact ggggccagat
     7081 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7141 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7201 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7261 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7321 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7381 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7441 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7501 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7561 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7621 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7681 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7741 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7801 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7861 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7921 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7981 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8041 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8101 cgcagcgagt cagtgagcga ggaagcggaa g
//
