LOCUS       sequence-180400-        8093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt41 promoter with Sp.his3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8093)
  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 8093)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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: pYZ166
FEATURES             Location/Qualifiers
     source          1..8093
                     /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..795
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISRLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFA
                     GFPRQALNRGLPLGFRFSALRHLDPKKLD"
     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      578..1033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(665..684)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     875..896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2771..3931
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     rep_origin      5029..5812
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6188..6292
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6293..6361,6362..7153)
                     /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(6511..6530)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7324..7912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7813..7832
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8066..8083
                     /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 tggcgcgacg cgccctgtag cggcgcatta
      601 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
      661 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      721 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      781 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      841 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
      901 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
      961 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     1021 acgtttacaa tttaaattag cgtaactcct ttacaaaatt aaaaataaca tcttttcatt
     1081 gcttatcgaa ctatacggaa acacaccaaa aaggaagaaa acgtatttta ataaagaaag
     1141 aaggaatgca agcgcaatta gcgaacacaa aatatattgg gtacatcaga aaagaaatgt
     1201 gacgagtccg gaaggggaaa ggaaagagtt ataatttgtc taattgaaac gttctaatag
     1261 gccagaaaag gaaacctcat ttttttgtat agtattcctg caatacaagt ttaatagttt
     1321 ttagtaatat cgtattctct tcttctgtgc caacagtgat acgcaatgct ccttcgcaaa
     1381 gaggctctgt acctctaaat ctaacaacga ctttgtgcat cgtggccatt tgtaaataaa
     1441 gatattttgc ggcatcgtta ctaggcttac ctccttctgg tctatctaaa acctgaatga
     1501 ggataaaatt tgcatcataa ccacctataa tcttgcccat tcctttaatg gtagtaagct
     1561 ctttgcaaag acgaactctt tgctgtataa tagcatctct atatgtatgc attttatcaa
     1621 tcgactgagg ggataaagca tccaaagcta aacgagaggt gggttcgcta atgttataag
     1681 gagcctttaa tgagttcata atggtagcaa taggtttact tgtcaaacaa aatccgatac
     1741 gaatacctgc tagaccgaaa gatttcgaaa gggtttggca aacagcaagg tttggatatt
     1801 cattgacaag agttaaggca gacatatcgg gagcagagaa atcaatatag gcctcatcga
     1861 cgacaacgat tccattccat gtagggtgct ccaaaatctt cttaatatcc tccagtttaa
     1921 gagccttagc tgttggatta ccaggggaac aagcaaaaaa gactttgatg gcactatctt
     1981 tagaaagtgt ttcgcaaata gcatcgacgt tcaaattgaa atctggttca aggagaacct
     2041 taacaacctc aacatcgttg atttttgcag acaccgtata cattccgtac gaaggtggac
     2101 acatcaaaat cttatctttt ccaggaatac aagaaatgcg aattaaggag tcaataattt
     2161 catcgctgcc aacacccatg caaatgttat ctggtgttag tggcttggta atggaaagct
     2221 ctttgtttct caaatcacat aatctttgct ttacttcgat ctgtctagga tcgggataac
     2281 gattaaattc cacaccgtcc acggaaataa ccgaaccata agcacattca ttggcatcaa
     2341 gaaggacacc ttcagagaag tcatctctcg cacagcgata aggctgaagt tctaagatat
     2401 ttttacgaag acaagtattc aaatcaaaca tcgtatctct ttcgatgtgt ttttgttaaa
     2461 gtaagaagaa agcgatcgat gagattaatt aataattaac ctaagatgca ggtgatatct
     2521 ataaatatat agtgtgtagc aatgtgagtc tttaattcat tacaacgtgt atcaccgtgt
     2581 cattatatcc aataaattca gaaacctgaa gaggaaagag aaataaagct cgatatgtct
     2641 ttcatttcca aatacaatta taagtgcaat agtaaagaaa acgttgaaag cttgataaaa
     2701 atttcgttac tatactaatg ctgtaaagaa aatgcaccat gtcgatcgac tctagaggat
     2761 cagaaaatta tcgccataaa agacagaata agtcatcagc ggttgtttca tttcctatat
     2821 ttttttttta tttttttatt ttttaataag ggaaaattta acgtctaagg atacagaaga
     2881 ttgttagcac attaaagtaa taaaggctta agtagtaagt gccttagcat gttattgtat
     2941 ttcaaaggac ataatctaaa ataataacaa tatcatttct cacaagttat tcaattttct
     3001 tttttttttc taataatatc aagaatgtat tatttgtttg acataagtca actaatttat
     3061 ttaatatgct ggattaatct tgcagacatg taaattaaca agttttagtc aaataacgtt
     3121 gaagtttcaa tgaactcaaa taatttctct ttttttttat ataaccataa gtctaatctg
     3181 atttatattt tccgcaggga tcaactgaag ttatgacatt tggattggat cacttataac
     3241 cttggtcgcc aaataataca aaaatcagcg ttataaaaca aagaaggttt ttgttaagaa
     3301 attaatcctc tttcttgata agaaagttga accgaaattg cagatactga tatatgaaaa
     3361 taatacccac aattttggga atagcgcaag cctcaattta aacaataggt gaggacacat
     3421 gataatgacc tcaatgattg ttagaagaaa agagcctcat tacaaaatcg aaaaatgaat
     3481 ggttgggtac aagtttccaa aacatggtaa agtggacttt gcgtatgaga cgtaaataga
     3541 aaaaaacact tgttatatgt tttctagaat tattgttgtc tctttatggt tggatgatgc
     3601 aaaatagtaa tttcggttag ttgctgtaaa acaccacgag acaaatagat atggatattt
     3661 attaaatcag gaaaaacgta actctcggct actggatggt tcagtcaccc aacgattact
     3721 ggggagagaa aacagggcaa aagcaaagct taaaggaatc cgattgtcat tcggcaatgt
     3781 gcagcgaaac taaaaaccgg ataatggacc tgttaatcga aacattgaag ataaaggaag
     3841 aggaatcctg gcatatcatc aattgaataa gttgaattaa ttatttcaat ctcattctca
     3901 ctttctgact tatagtcgct ttgttaaatc atatgtcgac tctagaggat ccccgggtaa
     3961 aaggaatgtc tcccttgcca gtactgctag ggtttttctt tcaaactatg gaagcccatt
     4021 caagctgcat attacgattt tgtttttcgc ttttagaaag tggtttagat gagataatag
     4081 aaaaattctt gatctccgac aacgagtact tttatttttt ttgctaatca ctttactcaa
     4141 tattagctcg aaatcgtaga aacgtagacg ggtgcgggat accgagtggt gtagttaaga
     4201 atttttataa accacgtggc ccaaaaatat gaacccaaaa cgtttataca tgagtatact
     4261 ttaagaaggc tatacccctt cgtgttagat gtagttttag ctacccaacc cgagtctatg
     4321 agcttgactt cagatgtaga aggcattaaa tcgttttgaa tattaattaa aaaacgatga
     4381 aaattaaata tttaaaagca atcatacgct gaaaatttag tgctgtggct aatccttcaa
     4441 catggaaatg ccataaaagt gactttgaca aaaaaaaaag tatatacagg tagtaaactc
     4501 atctacttca ttgactttgt ttacagcatg tggaaggagg aatatttatt gctaaatcgt
     4561 agtttaacat tcaataagta atactattga aattcgacaa gattggccgc atggatgaaa
     4621 aagaggcatt ttgctttggg agaattagtt caaattagaa ctgaaaaaaa aaactttacg
     4681 aggcaaaaat gtcggattga gatcgtaaaa gttcgctcgt cgtcttttgc tttgtgattg
     4741 ttttcatgga tacatcttgc tggatattta aattttagta ctatgtataa gatattctat
     4801 aaatgtttta tcacccaaac ctgttagcgc cttcttaatt ctattcaatc tggcttttgc
     4861 tctgagacta cttcttggac tttcactact tgttagttat acggaatttg tgtaattaga
     4921 agtgaaataa tcctttctat tagtaatgcg agctgcaatt cgagtctaac tccttaacca
     4981 ctcggacata gtgacttatc tgacactcat tgtaaattaa tatatatata ggcattttgt
     5041 ttagttaaag gtacttaagt aattagtata aacgaaccaa ttttataatc aggaagttaa
     5101 gtgaatggta gcacatgtcg taaaaattgt gaatttttat tgaataatat tttaaataca
     5161 agcctttcta gactaggtat actcataaac atatatgagc aaaaggatag aggagattac
     5221 attgcatctt ctacaaattt atttattgcc ctttactgaa aaattaaata atgagtacta
     5281 aatgataaaa agcgctcagt acaagaaaga ttgcaaaaat attgcattct tcatgaatta
     5341 aagttgcata taaggcatat tgaaagtaat agtactaaaa cagcagttag cgaaaattaa
     5401 tagaattata ttcgcaagac aattgtgaca ttaaattaaa aaattgtaga atttttacta
     5461 tcctctttaa cgccatgagc ctttataaaa aggttaaatt agttttaaca ttcttttttt
     5521 gagtaagagt ttacaattta tcaaaacctg tgttattata ttcattagtt tcaatttatt
     5581 agcatctaga gaaaaatcaa ttggcagtta cattgttgga atttatgaag aaaaagaatc
     5641 tacaacggag aataagttgc tgatcgcttt ccctaaaatt gtatattttg ctgagcttat
     5701 tttgacattt cgttgaagtt ttctctaatt cgcattcatt ttaagtaaaa caatgagaaa
     5761 taaaattaca aaaaatacaa attaaaatac aatttttagc tataattata gacgatgccc
     5821 ttgtatccca ttctgtctcg cttgccccta ctttttatct tttatatacc ataatgaacg
     5881 ctgccgctac taaccatacc ccgattttac atttcggact cccaaggacg tacaaaatag
     5941 aaaactatag aaaaaaataa tcagaaaata gcatgtcatc tctttgtaaa acgcgtttgc
     6001 aagaagaaag gaaacaatgg agaagagatc atccatttgt atgtaaattt tagtaaactt
     6061 gaagaaatca ctaacaactt ctcttactta gggattctat gcaaaacctt gtaaatcatc
     6121 tgatggagga ctcgatttaa tgaattggaa ggttggattt ccaggtggca cttttcgggg
     6181 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6241 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6301 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6361 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6421 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6481 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6541 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6601 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6661 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6721 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6781 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6841 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6901 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6961 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggat ctcgcggtat
     7021 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7081 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7141 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7201 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7261 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7321 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7381 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7441 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     7501 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7561 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7621 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7681 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga
     7741 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7801 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7861 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7921 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7981 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     8041 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
