LOCUS       sequence-199308-        6413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt1 promoter with empty BsaI-pad - no 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6413)
  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 6413)
  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: pYZ182
FEATURES             Location/Qualifiers
     source          1..6413
                     /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        1093..2251
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     rep_origin      3349..4132
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4508..4612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4613..4681,4682..5473)
                     /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(4831..4850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5644..6232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6133..6152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6386..6403
                     /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 gagaccggtt ctttggtcaa gaccgggtct ctgtcgatcg actctagagg
     1081 atcagaaaat tatcgccata aaagacagaa taagtcatca gcggttgttt catttcctat
     1141 attttttttt tattttttta ttttttaata agggaaaatt taacgtctaa ggatacagaa
     1201 gattgttagc acattaaagt aataaaggct taagtagtaa gtgccttagc atgttattgt
     1261 atttcaaagg acataatcta aaataataac aatatcattt ctcacaagtt attcaatttt
     1321 cttttttttt tctaataata tcaagaatgt attatttgtt tgacataagt caactaattt
     1381 atttaatatg ctggattaat cttgcagaca tgtaaattaa caagttttag tcaaataacg
     1441 ttgaagtttc aatgaactca aataatttct cttttttttt atataaccat aatctgattt
     1501 atattttccg cagggatcaa ctgaagttat gacatttgga ttggatcact tataaccttg
     1561 gtcgccaaat aatacaaaaa tcagcgttat aaaacaaaga aggtttttgt taagaaatta
     1621 atcctctttc ttgataagaa agttgaaccg aaattgcaga tactgatata tgaaaataat
     1681 acccacaatt ttgggaatag cgcaagcctc aatttaaaca ataggtgagg acacatgata
     1741 atgacctcaa tgattgttag aagaaaagag cctcattaca aaatcgaaaa atgaatggtt
     1801 gggtacaagt ttccaaaaca tggtaaagtg gactttgcgt atgagacgta aatagaaaaa
     1861 aacacttgtt atatgttttc tagaattatt gttgtctctt tatggttgga tgatgcaaaa
     1921 tagtaatttc ggttagttgc tgtaaaacac cacgagacaa atagatatgg atatttatta
     1981 aatcaggaaa aacgtaactc tcggctactg gatggttcag tcacccaacg attactgggg
     2041 agagaaaaca gggcaaaagc aaagcttaaa ggaatccgat tgtcattcgg caatgtgcag
     2101 cgaaactaaa aaccggataa tggacctgtt aatcgaaaca ttgaagatat ataaaggaag
     2161 aggaatcctg gcatatcatc aattgaataa gttgaattaa ttatttcaat ctcattctca
     2221 ctttctgact tatagtcgct ttgttaaatc atatgtcgac tctagaggat ccccgggtaa
     2281 aaggaatgtc tcccttgcca gtactgctag ggtttttctt tcaaactatg gaagcccatt
     2341 caagctgcat attacgattt tgtttttcgc ttttagaaag tggtttagat gagataatag
     2401 aaaaattctt gatctccgac aacgagtact tttatttttt ttgctaatca ctttactcaa
     2461 tattagctcg aaatcgtaga aacgtagacg ggtgcgggat accgagtggt gtagttaaga
     2521 atttttataa accacgtggc ccaaaaatat gaacccaaaa cgtttataca tgagtatact
     2581 ttaagaaggc tatacccctt cgtgttagat gtagttttag ctacccaacc cgagtctatg
     2641 agcttgactt cagatgtaga aggcattaaa tcgttttgaa tattaattaa aaaacgatga
     2701 aaattaaata tttaaaagca atcatacgct gaaaatttag tgctgtggct aatccttcaa
     2761 catggaaatg ccataaaagt gactttgaca aaaaaaaaag tatatacagg tagtaaactc
     2821 atctacttca ttgactttgt ttacagcatg tggaaggagg aatatttatt gctaaatcgt
     2881 agtttaacat tcaataagta atactattga aattcgacaa gattggccgc atggatgaaa
     2941 aagaggcatt ttgctttggg agaattagtc caaattagaa ctgaaaaaaa aaactttacg
     3001 aggcaaaaat gtcggattga gatcgtaaaa gttcgctcgt cgtcttttgc tttgtgattg
     3061 ttttcatgga tacatcttgc tggatattta aattttagta ctatgtataa gatattctat
     3121 aaatgtttta tcacccaaac ctgttagcgc cttcttaatt ctattcaatc tggcttttgc
     3181 tctgagacta cttcttggac tttcactact tgttagttat acggaatttg tgtaattaga
     3241 agtgaaataa tcctttctat tagtaatgcg agctgcaatt cgagtctaac tccttaacca
     3301 ctcggacata gtgacttatc tgacactcat tgtaaattaa tatatatata ggcattttgt
     3361 ttagttaaag gtacttaagt aattagtata aacgaaccaa ttttataatc aggaagttaa
     3421 gtgaatggta gcacatgtcg taaaaattgt gaatttttat tgaataatat tttaaataca
     3481 agcctttcta gactaggtat actcataaac atatatgagc aaaaggatag aggagattac
     3541 attgcatctt ctacaaattt atttattgcc ctttactgaa aaattaaata atgagtacta
     3601 aatgataaaa agcgctcagt acaagaaaga ttgcaaaaat attgcattct tcatgaatta
     3661 aagttgcata taaggcatat tgaaagtaat agtactaaaa cagcagttag cgaaaattaa
     3721 tagaattata ttcgcaagac aattgtgaca ttaaattaaa aaattgtaga atttttacta
     3781 tcctctttaa cgccatgagc ctttataaaa aggttaaatt agttttaaca ttcttttttt
     3841 gagtaagagt ttacaattta tcaaaacctg tgttattata ttcattagtt tcaatttatt
     3901 agcatctaga gaaaaatcaa ttggcagtta cattgttgga atttatgaag aaaaagaatc
     3961 tacaacggag aataagttgc tgatcgcttt ccctaaaatt gtatattttg ctgagcttat
     4021 tttgacattt cgttgaagtt ttctctaatt cgcattcatt ttaagtaaaa caatgagaaa
     4081 taaaattaca aaaaatacaa attaaaatac aatttttagc tataattata gacgatgccc
     4141 ttgtatccca ttctgtctcg cttgccccta ctttttatct tttatatacc ataatgaacg
     4201 ctgccgctac taaccatacc ccgattttac atttcggact cccaaggacg tacaaaatag
     4261 aaaactatag aaaaaaataa tcagaaaata gcatgtcatc tctttgtaaa acgcgtttgc
     4321 aagaagaaag gaaacaatgg agaagagatc atccatttgt atgtaaattt tagtaaactt
     4381 gaagaaatca ctaacaactt ctcttactta gggattctat gcaaaacctt gtaaatcatc
     4441 tgatggagga ctcgatttaa tgaattggaa ggttggattt ccaggtggca cttttcgggg
     4501 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4561 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4621 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4681 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4741 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4801 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4861 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4921 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4981 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5041 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5101 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5161 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5221 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5281 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggat ctcgcggtat
     5341 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5401 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5461 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5521 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5581 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5641 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5701 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5761 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5821 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5881 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5941 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6001 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6061 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
