LOCUS       sequence-207093-        8087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt1 promoter with Sp.his3 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8087)
  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 8087)
  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: pYZ165
FEATURES             Location/Qualifiers
     source          1..8087
                     /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..3925
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     rep_origin      5023..5806
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6182..6286
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6287..6355,6356..7147)
                     /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(6505..6524)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7318..7906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7807..7826
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8060..8077
                     /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 atatataaag gaagaggaat
     3841 cctggcatat catcaattga ataagttgaa ttaattattt caatctcatt ctcactttct
     3901 gacttatagt cgctttgtta aatcatatgt cgactctaga ggatccccgg gtaaaaggaa
     3961 tgtctccctt gccagtactg ctagggtttt tctttcaaac tatggaagcc cattcaagct
     4021 gcatattacg attttgtttt tcgcttttag aaagtggttt agatgagata atagaaaaat
     4081 tcttgatctc cgacaacgag tacttttatt ttttttgcta atcactttac tcaatattag
     4141 ctcgaaatcg tagaaacgta gacgggtgcg ggataccgag tggtgtagtt aagaattttt
     4201 ataaaccacg tggcccaaaa atatgaaccc aaaacgttta tacatgagta tactttaaga
     4261 aggctatacc ccttcgtgtt agatgtagtt ttagctaccc aacccgagtc tatgagcttg
     4321 acttcagatg tagaaggcat taaatcgttt tgaatattaa ttaaaaaacg atgaaaatta
     4381 aatatttaaa agcaatcata cgctgaaaat ttagtgctgt ggctaatcct tcaacatgga
     4441 aatgccataa aagtgacttt gacaaaaaaa aaagtatata caggtagtaa actcatctac
     4501 ttcattgact ttgtttacag catgtggaag gaggaatatt tattgctaaa tcgtagttta
     4561 acattcaata agtaatacta ttgaaattcg acaagattgg ccgcatggat gaaaaagagg
     4621 cattttgctt tgggagaatt agtccaaatt agaactgaaa aaaaaaactt tacgaggcaa
     4681 aaatgtcgga ttgagatcgt aaaagttcgc tcgtcgtctt ttgctttgtg attgttttca
     4741 tggatacatc ttgctggata tttaaatttt agtactatgt ataagatatt ctataaatgt
     4801 tttatcaccc aaacctgtta gcgccttctt aattctattc aatctggctt ttgctctgag
     4861 actacttctt ggactttcac tacttgttag ttatacggaa tttgtgtaat tagaagtgaa
     4921 ataatccttt ctattagtaa tgcgagctgc aattcgagtc taactcctta accactcgga
     4981 catagtgact tatctgacac tcattgtaaa ttaatatata tataggcatt ttgtttagtt
     5041 aaaggtactt aagtaattag tataaacgaa ccaattttat aatcaggaag ttaagtgaat
     5101 ggtagcacat gtcgtaaaaa ttgtgaattt ttattgaata atattttaaa tacaagcctt
     5161 tctagactag gtatactcat aaacatatat gagcaaaagg atagaggaga ttacattgca
     5221 tcttctacaa atttatttat tgccctttac tgaaaaatta aataatgagt actaaatgat
     5281 aaaaagcgct cagtacaaga aagattgcaa aaatattgca ttcttcatga attaaagttg
     5341 catataaggc atattgaaag taatagtact aaaacagcag ttagcgaaaa ttaatagaat
     5401 tatattcgca agacaattgt gacattaaat taaaaaattg tagaattttt actatcctct
     5461 ttaacgccat gagcctttat aaaaaggtta aattagtttt aacattcttt ttttgagtaa
     5521 gagtttacaa tttatcaaaa cctgtgttat tatattcatt agtttcaatt tattagcatc
     5581 tagagaaaaa tcaattggca gttacattgt tggaatttat gaagaaaaag aatctacaac
     5641 ggagaataag ttgctgatcg ctttccctaa aattgtatat tttgctgagc ttattttgac
     5701 atttcgttga agttttctct aattcgcatt cattttaagt aaaacaatga gaaataaaat
     5761 tacaaaaaat acaaattaaa atacaatttt tagctataat tatagacgat gcccttgtat
     5821 cccattctgt ctcgcttgcc cctacttttt atcttttata taccataatg aacgctgccg
     5881 ctactaacca taccccgatt ttacatttcg gactcccaag gacgtacaaa atagaaaact
     5941 atagaaaaaa ataatcagaa aatagcatgt catctctttg taaaacgcgt ttgcaagaag
     6001 aaaggaaaca atggagaaga gatcatccat ttgtatgtaa attttagtaa acttgaagaa
     6061 atcactaaca acttctctta cttagggatt ctatgcaaaa ccttgtaaat catctgatgg
     6121 aggactcgat ttaatgaatt ggaaggttgg atttccaggt ggcacttttc ggggaaatgt
     6181 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6241 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6301 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6361 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6421 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6481 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6541 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6601 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6661 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6721 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6781 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6841 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6901 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6961 tggctggttt attgctgata aatctggagc cggtgagcgt ggatctcgcg gtatcattgc
     7021 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7081 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7141 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7201 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7261 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7321 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7381 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7441 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7501 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7561 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7621 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7681 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7741 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7801 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7861 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7921 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7981 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     8041 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
