LOCUS       sequence-180401-        8090 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 8090)
  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 8090)
  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..8090
                     /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..3928
                     /label=nmt1 P81 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring weak thiamine-repressible expression"
     rep_origin      5026..5809
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6185..6289
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6290..6358,6359..7150)
                     /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(6508..6527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7321..7909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7810..7829
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8063..8080
                     /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 atggaagagg
     3841 aatcctggca tatcatcaat tgaataagtt gaattaatta tttcaatctc attctcactt
     3901 tctgacttat agtcgctttg ttaaatcata tgtcgactct agaggatccc cgggtaaaag
     3961 gaatgtctcc cttgccagta ctgctagggt ttttctttca aactatggaa gcccattcaa
     4021 gctgcatatt acgattttgt ttttcgcttt tagaaagtgg tttagatgag ataatagaaa
     4081 aattcttgat ctccgacaac gagtactttt attttttttg ctaatcactt tactcaatat
     4141 tagctcgaaa tcgtagaaac gtagacgggt gcgggatacc gagtggtgta gttaagaatt
     4201 tttataaacc acgtggccca aaaatatgaa cccaaaacgt ttatacatga gtatacttta
     4261 agaaggctat accccttcgt gttagatgta gttttagcta cccaacccga gtctatgagc
     4321 ttgacttcag atgtagaagg cattaaatcg ttttgaatat taattaaaaa acgatgaaaa
     4381 ttaaatattt aaaagcaatc atacgctgaa aatttagtgc tgtggctaat ccttcaacat
     4441 ggaaatgcca taaaagtgac tttgacaaaa aaaaaagtat atacaggtag taaactcatc
     4501 tacttcattg actttgttta cagcatgtgg aaggaggaat atttattgct aaatcgtagt
     4561 ttaacattca ataagtaata ctattgaaat tcgacaagat tggccgcatg gatgaaaaag
     4621 aggcattttg ctttgggaga attagttcaa attagaactg aaaaaaaaaa ctttacgagg
     4681 caaaaatgtc ggattgagat cgtaaaagtt cgctcgtcgt cttttgcttt gtgattgttt
     4741 tcatggatac atcttgctgg atatttaaat tttagtacta tgtataagat attctataaa
     4801 tgttttatca cccaaacctg ttagcgcctt cttaattcta ttcaatctgg cttttgctct
     4861 gagactactt cttggacttt cactacttgt tagttatacg gaatttgtgt aattagaagt
     4921 gaaataatcc tttctattag taatgcgagc tgcaattcga gtctaactcc ttaaccactc
     4981 ggacatagtg acttatctga cactcattgt aaattaatat atatataggc attttgttta
     5041 gttaaaggta cttaagtaat tagtataaac gaaccaattt tataatcagg aagttaagtg
     5101 aatggtagca catgtcgtaa aaattgtgaa tttttattga ataatatttt aaatacaagc
     5161 ctttctagac taggtatact cataaacata tatgagcaaa aggatagagg agattacatt
     5221 gcatcttcta caaatttatt tattgccctt tactgaaaaa ttaaataatg agtactaaat
     5281 gataaaaagc gctcagtaca agaaagattg caaaaatatt gcattcttca tgaattaaag
     5341 ttgcatataa ggcatattga aagtaatagt actaaaacag cagttagcga aaattaatag
     5401 aattatattc gcaagacaat tgtgacatta aattaaaaaa ttgtagaatt tttactatcc
     5461 tctttaacgc catgagcctt tataaaaagg ttaaattagt tttaacattc tttttttgag
     5521 taagagttta caatttatca aaacctgtgt tattatattc attagtttca atttattagc
     5581 atctagagaa aaatcaattg gcagttacat tgttggaatt tatgaagaaa aagaatctac
     5641 aacggagaat aagttgctga tcgctttccc taaaattgta tattttgctg agcttatttt
     5701 gacatttcgt tgaagttttc tctaattcgc attcatttta agtaaaacaa tgagaaataa
     5761 aattacaaaa aatacaaatt aaaatacaat ttttagctat aattatagac gatgcccttg
     5821 tatcccattc tgtctcgctt gcccctactt tttatctttt atataccata atgaacgctg
     5881 ccgctactaa ccataccccg attttacatt tcggactccc aaggacgtac aaaatagaaa
     5941 actatagaaa aaaataatca gaaaatagca tgtcatctct ttgtaaaacg cgtttgcaag
     6001 aagaaaggaa acaatggaga agagatcatc catttgtatg taaattttag taaacttgaa
     6061 gaaatcacta acaacttctc ttacttaggg attctatgca aaaccttgta aatcatctga
     6121 tggaggactc gatttaatga attggaaggt tggatttcca ggtggcactt ttcggggaaa
     6181 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6241 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6301 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6361 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6421 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6481 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6541 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6601 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6661 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6721 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6781 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6841 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6901 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6961 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggatctc gcggtatcat
     7021 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7081 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7141 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7201 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7261 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7321 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7381 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7441 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     7501 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7561 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7621 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7681 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     7741 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7801 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7861 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7921 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7981 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8041 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
