LOCUS       sequence-199356-        8080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt81 promoter with Sp.his3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8080)
  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 8080)
  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: pYZ167
FEATURES             Location/Qualifiers
     source          1..8080
                     /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..3918
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     rep_origin      5016..5799
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6175..6279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6280..6348,6349..7140)
                     /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(6498..6517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7311..7899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7800..7819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8053..8070
                     /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 atggaagagg aatcctggca
     3841 tatcatcaat tgaataagtt gaattaatta tttcaatctc attctcactt tctgacttat
     3901 agtcgctttg ttaaatcata tgtcgactct agaggatccc cgggtaaaag gaatgtctcc
     3961 cttgccagta ctgctagggt ttttctttca aactatggaa gcccattcaa gctgcatatt
     4021 acgattttgt ttttcgcttt tagaaagtgg tttagatgag ataatagaaa aattcttgat
     4081 ctccgacaac gagtactttt attttttttg ctaatcactt tactcaatat tagctcgaaa
     4141 tcgtagaaac gtagacgggt gcgggatacc gagtggtgta gttaagaatt tttataaacc
     4201 acgtggccca aaaatatgaa cccaaaacgt ttatacatga gtatacttta agaaggctat
     4261 accccttcgt gttagatgta gttttagcta cccaacccga gtctatgagc ttgacttcag
     4321 atgtagaagg cattaaatcg ttttgaatat taattaaaaa acgatgaaaa ttaaatattt
     4381 aaaagcaatc atacgctgaa aatttagtgc tgtggctaat ccttcaacat ggaaatgcca
     4441 taaaagtgac tttgacaaaa aaaaaagtat atacaggtag taaactcatc tacttcattg
     4501 actttgttta cagcatgtgg aaggaggaat atttattgct aaatcgtagt ttaacattca
     4561 ataagtaata ctattgaaat tcgacaagat tggccgcatg gatgaaaaag aggcattttg
     4621 ctttgggaga attagtccaa attagaactg aaaaaaaaaa ctttacgagg caaaaatgtc
     4681 ggattgagat cgtaaaagtt cgctcgtcgt cttttgcttt gtgattgttt tcatggatac
     4741 atcttgctgg atatttaaat tttagtacta tgtataagat attctataaa tgttttatca
     4801 cccaaacctg ttagcgcctt cttaattcta ttcaatctgg cttttgctct gagactactt
     4861 cttggacttt cactacttgt tagttatacg gaatttgtgt aattagaagt gaaataatcc
     4921 tttctattag taatgcgagc tgcaattcga gtctaactcc ttaaccactc ggacatagtg
     4981 acttatctga cactcattgt aaattaatat atatataggc attttgttta gttaaaggta
     5041 cttaagtaat tagtataaac gaaccaattt tataatcagg aagttaagtg aatggtagca
     5101 catgtcgtaa aaattgtgaa tttttattga ataatatttt aaatacaagc ctttctagac
     5161 taggtatact cataaacata tatgagcaaa aggatagagg agattacatt gcatcttcta
     5221 caaatttatt tattgccctt tactgaaaaa ttaaataatg agtactaaat gataaaaagc
     5281 gctcagtaca agaaagattg caaaaatatt gcattcttca tgaattaaag ttgcatataa
     5341 ggcatattga aagtaatagt actaaaacag cagttagcga aaattaatag aattatattc
     5401 gcaagacaat tgtgacatta aattaaaaaa ttgtagaatt tttactatcc tctttaacgc
     5461 catgagcctt tataaaaagg ttaaattagt tttaacattc tttttttgag taagagttta
     5521 caatttatca aaacctgtgt tattatattc attagtttca atttattagc atctagagaa
     5581 aaatcaattg gcagttacat tgttggaatt tatgaagaaa aagaatctac aacggagaat
     5641 aagttgctga tcgctttccc taaaattgta tattttgctg agcttatttt gacatttcgt
     5701 tgaagttttc tctaattcgc attcatttta agtaaaacaa tgagaaataa aattacaaaa
     5761 aatacaaatt aaaatacaat ttttagctat aattatagac gatgcccttg tatcccattc
     5821 tgtctcgctt gcccctactt tttatctttt atataccata atgaacgctg ccgctactaa
     5881 ccataccccg attttacatt tcggactccc aaggacgtac aaaatagaaa actatagaaa
     5941 aaaataatca gaaaatagca tgtcatctct ttgtaaaacg cgtttgcaag aagaaaggaa
     6001 acaatggaga agagatcatc catttgtatg taaattttag taaacttgaa gaaatcacta
     6061 acaacttctc ttacttaggg attctatgca aaaccttgta aatcatctga tggaggactc
     6121 gatttaatga attggaaggt tggatttcca ggtggcactt ttcggggaaa tgtgcgcgga
     6181 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6241 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6301 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6361 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6421 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6481 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6541 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6601 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6661 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6721 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6781 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6841 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6901 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6961 tttattgctg ataaatctgg agccggtgag cgtggatctc gcggtatcat tgcagcactg
     7021 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7081 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7141 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7201 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7261 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7321 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7381 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7441 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7501 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7561 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7621 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7681 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7741 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7801 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7861 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7921 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7981 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     8041 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
