LOCUS       sequence-180399-        8097 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 8097)
  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 8097)
  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..8097
                     /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..3935
                     /label=nmt1 promoter
                     /note="wild-type nmt1 promoter from Schizosaccharomyces 
                     pombe, conferring strong thiamine-repressible expression"
     rep_origin      5033..5816
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        6192..6296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6297..6365,6366..7157)
                     /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(6515..6534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7328..7916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7817..7836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8070..8087
                     /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 atatataaag
     3841 gaagaggaat cctggcatat catcaattga ataagttgaa ttaattattt caatctcatt
     3901 ctcactttct gacttatagt cgctttgtta aatcatatgt cgactctaga ggatccccgg
     3961 gtaaaaggaa tgtctccctt gccagtactg ctagggtttt tctttcaaac tatggaagcc
     4021 cattcaagct gcatattacg attttgtttt tcgcttttag aaagtggttt agatgagata
     4081 atagaaaaat tcttgatctc cgacaacgag tacttttatt ttttttgcta atcactttac
     4141 tcaatattag ctcgaaatcg tagaaacgta gacgggtgcg ggataccgag tggtgtagtt
     4201 aagaattttt ataaaccacg tggcccaaaa atatgaaccc aaaacgttta tacatgagta
     4261 tactttaaga aggctatacc ccttcgtgtt agatgtagtt ttagctaccc aacccgagtc
     4321 tatgagcttg acttcagatg tagaaggcat taaatcgttt tgaatattaa ttaaaaaacg
     4381 atgaaaatta aatatttaaa agcaatcata cgctgaaaat ttagtgctgt ggctaatcct
     4441 tcaacatgga aatgccataa aagtgacttt gacaaaaaaa aaagtatata caggtagtaa
     4501 actcatctac ttcattgact ttgtttacag catgtggaag gaggaatatt tattgctaaa
     4561 tcgtagttta acattcaata agtaatacta ttgaaattcg acaagattgg ccgcatggat
     4621 gaaaaagagg cattttgctt tgggagaatt agttcaaatt agaactgaaa aaaaaaactt
     4681 tacgaggcaa aaatgtcgga ttgagatcgt aaaagttcgc tcgtcgtctt ttgctttgtg
     4741 attgttttca tggatacatc ttgctggata tttaaatttt agtactatgt ataagatatt
     4801 ctataaatgt tttatcaccc aaacctgtta gcgccttctt aattctattc aatctggctt
     4861 ttgctctgag actacttctt ggactttcac tacttgttag ttatacggaa tttgtgtaat
     4921 tagaagtgaa ataatccttt ctattagtaa tgcgagctgc aattcgagtc taactcctta
     4981 accactcgga catagtgact tatctgacac tcattgtaaa ttaatatata tataggcatt
     5041 ttgtttagtt aaaggtactt aagtaattag tataaacgaa ccaattttat aatcaggaag
     5101 ttaagtgaat ggtagcacat gtcgtaaaaa ttgtgaattt ttattgaata atattttaaa
     5161 tacaagcctt tctagactag gtatactcat aaacatatat gagcaaaagg atagaggaga
     5221 ttacattgca tcttctacaa atttatttat tgccctttac tgaaaaatta aataatgagt
     5281 actaaatgat aaaaagcgct cagtacaaga aagattgcaa aaatattgca ttcttcatga
     5341 attaaagttg catataaggc atattgaaag taatagtact aaaacagcag ttagcgaaaa
     5401 ttaatagaat tatattcgca agacaattgt gacattaaat taaaaaattg tagaattttt
     5461 actatcctct ttaacgccat gagcctttat aaaaaggtta aattagtttt aacattcttt
     5521 ttttgagtaa gagtttacaa tttatcaaaa cctgtgttat tatattcatt agtttcaatt
     5581 tattagcatc tagagaaaaa tcaattggca gttacattgt tggaatttat gaagaaaaag
     5641 aatctacaac ggagaataag ttgctgatcg ctttccctaa aattgtatat tttgctgagc
     5701 ttattttgac atttcgttga agttttctct aattcgcatt cattttaagt aaaacaatga
     5761 gaaataaaat tacaaaaaat acaaattaaa atacaatttt tagctataat tatagacgat
     5821 gcccttgtat cccattctgt ctcgcttgcc cctacttttt atcttttata taccataatg
     5881 aacgctgccg ctactaacca taccccgatt ttacatttcg gactcccaag gacgtacaaa
     5941 atagaaaact atagaaaaaa ataatcagaa aatagcatgt catctctttg taaaacgcgt
     6001 ttgcaagaag aaaggaaaca atggagaaga gatcatccat ttgtatgtaa attttagtaa
     6061 acttgaagaa atcactaaca acttctctta cttagggatt ctatgcaaaa ccttgtaaat
     6121 catctgatgg aggactcgat ttaatgaatt ggaaggttgg atttccaggt ggcacttttc
     6181 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6241 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6301 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6361 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6421 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6481 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6541 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6601 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6661 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6721 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6781 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6841 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6901 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6961 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggatctcgcg
     7021 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7081 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7141 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7201 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7261 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7321 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7381 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7441 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     7501 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7561 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7621 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7681 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     7741 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7801 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7861 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7921 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7981 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8041 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
