LOCUS       sequence-198986-        8082 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 8082)
  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 8082)
  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..8082
                     /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"
     promoter        2767..3921
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     rep_origin      5018..5801
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6177..6281
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6282..6350,6351..7142)
                     /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(6500..6519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7313..7901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7802..7821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8055..8072
                     /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 aattagcgta actcctttac aaaattaaaa ataacatctt ttcattgctt
     1081 atcgaactat acggaaacac accaaaaagg aagaaaacgt attttaataa agaaagaagg
     1141 aatgcaagcg caattagcga acacaaaata tattgggtac atcagaaaag aaatgtgacg
     1201 agtccggaag gggaaaggaa agagttataa tttgtctaat tgaaacgttc taataggcca
     1261 gaaaaggaaa cctcattttt ttgtatagta ttcctgcaat acaagtttaa tagtttttag
     1321 taatatcgta ttctcttctt ctgtgccaac agtgatacgc aatgctcctt cgcaaagagg
     1381 ctctgtacct ctaaatctaa caacgacttt gtgcatcgtg gccatttgta aataaagata
     1441 ttttgcggca tcgttactag gcttacctcc ttctggtcta tctaaaacct gaatgaggat
     1501 aaaatttgca tcataaccac ctataatctt gcccattcct ttaatggtag taagctcttt
     1561 gcaaagacga actctttgct gtataatagc atctctatat gtatgcattt tatcaatcga
     1621 ctgaggggat aaagcatcca aagctaaacg agaggtgggt tcgctaatgt tataaggagc
     1681 ctttaatgag ttcataatgg tagcaatagg tttacttgtc aaacaaaatc cgatacgaat
     1741 acctgctaga ccgaaagatt tcgaaagggt ttggcaaaca gcaaggtttg gatattcatt
     1801 gacaagagtt aaggcagaca tatcgggagc agagaaatca atataggcct catcgacgac
     1861 aacgattcca ttccatgtag ggtgctccaa aatcttctta atatcctcca gtttaagagc
     1921 cttagctgtt ggattaccag gggaacaagc aaaaaagact ttgatggcac tatctttaga
     1981 aagtgtttcg caaatagcat cgacgttcaa attgaaatct ggttcaagga gaaccttaac
     2041 aacctcaaca tcgttgattt ttgcagacac cgtatacatt ccgtacgaag gtggacacat
     2101 caaaatctta tcttttccag gaatacaaga aatgcgaatt aaggagtcaa taatttcatc
     2161 gctgccaaca cccatgcaaa tgttatctgg tgttagtggc ttggtaatgg aaagctcttt
     2221 gtttctcaaa tcacataatc tttgctttac ttcgatctgt ctaggatcgg gataacgatt
     2281 aaattccaca ccgtccacgg aaataaccga accataagca cattcattgg catcaagaag
     2341 gacaccttca gagaagtcat ctctcgcaca gcgataaggc tgaagttcta agatattttt
     2401 acgaagacaa gtattcaaat caaacatcgt atctctttcg atgtgttttt gttaaagtaa
     2461 gaagaaagcg atcgatgaga ttaattaata attaacctaa gatgcaggtg atatctataa
     2521 atatatagtg tgtagcaatg tgagtcttta attcattaca acgtgtatca ccgtgtcatt
     2581 atatccaata aattcagaaa cctgaagagg aaagagaaat aaagctcgat atgtctttca
     2641 tttccaaata caattataag tgcaatagta aagaaaacgt tgaaagcttg ataaaaattt
     2701 cgttactata ctaatgctgt aaagaaaatg caccatgtcg atcgactcta gaggatcaga
     2761 aaattatcgc cataaaagac agaataagtc atcagcggtt gtttcatttc ctatattttt
     2821 tttttatttt tttatttttt aataagggaa aatttaacgt ctaaggatac agaagattgt
     2881 tagcacatta aagtaataaa ggcttaagta gtaagtgcct tagcatgtta ttgtatttca
     2941 aaggacataa tctaaaataa taacaatatc atttctcaca agttattcaa ttttcttttt
     3001 tttttctaat aatatcaaga atgtattatt tgtttgacat aagtcaacta atttatttaa
     3061 tatgctggat taatcttgca gacatgtaaa ttaacaagtt ttagtcaaat aacgttgaag
     3121 tttcaatgaa ctcaaataat ttctcttttt ttttatataa ccataatctg atttatattt
     3181 tccgcaggga tcaactgaag ttatgacatt tggattggat cacttataac cttggtcgcc
     3241 aaataataca aaaatcagcg ttataaaaca aagaaggttt ttgttaagaa attaatcctc
     3301 tttcttgata agaaagttga accgaaattg cagatactga tatatgaaaa taatacccac
     3361 aattttggga atagcgcaag cctcaattta aacaataggt gaggacacat gataatgacc
     3421 tcaatgattg ttagaagaaa agagcctcat tacaaaatcg aaaaatgaat ggttgggtac
     3481 aagtttccaa aacatggtaa agtggacttt gcgtatgaga cgtaaataga aaaaaacact
     3541 tgttatatgt tttctagaat tattgttgtc tctttatggt tggatgatgc aaaatagtaa
     3601 tttcggttag ttgctgtaaa acaccacgag acaaatagat atggatattt attaaatcag
     3661 gaaaaacgta actctcggct actggatggt tcagtcaccc aacgattact ggggagagaa
     3721 aacagggcaa aagcaaagct taaaggaatc cgattgtcat tcggcaatgt gcagcgaaac
     3781 taaaaaccgg ataatggacc tgttaatcga aacattgaag ataaaggaag aggaatcctg
     3841 gcatatcatc aattgaataa gttgaattaa ttatttcaat ctcattctca ctttctgact
     3901 tatagtcgct ttgttaaatc atatgtcgac tctagaggat ccccgggtaa aaggaatgtc
     3961 tcccttgcca gtactgctag ggtttttctt tcaaactatg gaagcccatt caagctgcat
     4021 attacgattt tgtttttcgc ttttagaaag tggtttagat gagataatag aaaaattctt
     4081 gatctccgac aacgagtact tttatttttt ttgctaatca ctttactcaa tattagctcg
     4141 aaatcgtaga aacgtagacg ggtgcgggat accgagtggt gtagttaaga atttttataa
     4201 accacgtggc ccaaaaatat gaacccaaaa cgtttataca tgagtatact ttaagaaggc
     4261 tatacccctt cgtgttagat gtagttttag ctacccaacc cgagtctatg agcttgactt
     4321 cagatgtaga aggcattaaa tcgttttgaa tattaattaa aaaacgatga aaattaaata
     4381 tttaaaagca atcatacgct gaaaatttag tgctgtggct aatccttcaa catggaaatg
     4441 ccataaaagt gactttgaca aaaaaaaaag tatatacagg tagtaaactc atctacttca
     4501 ttgactttgt ttacagcatg tggaaggagg aatatttatt gctaaatcgt agtttaacat
     4561 tcaataagta atactattga aattcgacaa gattggccgc atggatgaaa aagaggcatt
     4621 ttgctttggg agaattagtc caaattagaa ctgaaaaaaa aactttacga ggcaaaaatg
     4681 tcggattgag atcgtaaaag ttcgctcgtc gtcttttgct ttgtgattgt tttcatggat
     4741 acatcttgct ggatatttaa attttagtac tatgtataag atattctata aatgttttat
     4801 cacccaaacc tgttagcgcc ttcttaattc tattcaatct ggcttttgct ctgagactac
     4861 ttcttggact ttcactactt gttagttata cggaatttgt gtaattagaa gtgaaataat
     4921 cctttctatt agtaatgcga gctgcaattc gagtctaact ccttaaccac tcggacatag
     4981 tgacttatct gacactcatt gtaaattaat atatatatag gcattttgtt tagttaaagg
     5041 tacttaagta attagtataa acgaaccaat tttataatca ggaagttaag tgaatggtag
     5101 cacatgtcgt aaaaattgtg aatttttatt gaataatatt ttaaatacaa gcctttctag
     5161 actaggtata ctcataaaca tatatgagca aaaggataga ggagattaca ttgcatcttc
     5221 tacaaattta tttattgccc tttactgaaa aattaaataa tgagtactaa atgataaaaa
     5281 gcgctcagta caagaaagat tgcaaaaata ttgcattctt catgaattaa agttgcatat
     5341 aaggcatatt gaaagtaata gtactaaaac agcagttagc gaaaattaat agaattatat
     5401 tcgcaagaca attgtgacat taaattaaaa aattgtagaa tttttactat cctctttaac
     5461 gccatgagcc tttataaaaa ggttaaatta gttttaacat tctttttttg agtaagagtt
     5521 tacaatttat caaaacctgt gttattatat tcattagttt caatttatta gcatctagag
     5581 aaaaatcaat tggcagttac attgttggaa tttatgaaga aaaagaatct acaacggaga
     5641 ataagttgct gatcgctttc cctaaaattg tatattttgc tgagcttatt ttgacatttc
     5701 gttgaagttt tctctaattc gcattcattt taagtaaaac aatgagaaat aaaattacaa
     5761 aaaatacaaa ttaaaataca atttttagct ataattatag acgatgccct tgtatcccat
     5821 tctgtctcgc ttgcccctac tttttatctt ttatatacca taatgaacgc tgccgctact
     5881 aaccataccc cgattttaca tttcggactc ccaaggacgt acaaaataga aaactataga
     5941 aaaaaataat cagaaaatag catgtcatct ctttgtaaaa cgcgtttgca agaagaaagg
     6001 aaacaatgga gaagagatca tccatttgta tgtaaatttt agtaaacttg aagaaatcac
     6061 taacaacttc tcttacttag ggattctatg caaaaccttg taaatcatct gatggaggac
     6121 tcgatttaat gaattggaag gttggatttc caggtggcac ttttcgggga aatgtgcgcg
     6181 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6241 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6301 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6361 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6421 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6481 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6541 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6601 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6661 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6721 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6781 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6841 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6901 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6961 ggtttattgc tgataaatct ggagccggtg agcgtggatc tcgcggtatc attgcagcac
     7021 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7081 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     7141 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     7201 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7261 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7321 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7381 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7441 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7501 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7561 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7621 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7681 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7741 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7801 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7861 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7921 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7981 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     8041 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//
