LOCUS       sequence-180406-        6420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pombe Expression Vector - nmt41 promoter with empty BsaI-pad - no 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6420)
  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 6420)
  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: pYZ183
FEATURES             Location/Qualifiers
     source          1..6420
                     /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        1098..2258
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     rep_origin      3356..4139
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        4515..4619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4620..4688,4689..5480)
                     /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(4838..4857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5651..6239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6140..6159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6393..6410
                     /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 tttagagacc ggttctttgg gtcaagaccg ggtctctgtc gatcgactct
     1081 agaggatcag aaaattatcg ccataaaaga cagaataagt catcagcggt tgtttcattt
     1141 cctatatttt ttttttattt ttttattttt taataaggga aaatttaacg tctaaggata
     1201 cagaagattg ttagcacatt aaagtaataa aggcttaagt agtaagtgcc ttagcatgtt
     1261 attgtatttc aaaggacata atctaaaata ataacaatat catttctcac aagttattca
     1321 attttctttt ttttttctaa taatatcaag aatgtattat ttgtttgaca taagtcaact
     1381 aatttattta atatgctgga ttaatcttgc agacatgtaa attaacaagt tttagtcaaa
     1441 taacgttgaa gtttcaatga actcaaataa tttctctttt tttttatata accataagtc
     1501 taatctgatt tatattttcc gcagggatca actgaagtta tgacatttgg attggatcac
     1561 ttataacctt ggtcgccaaa taatacaaaa atcagcgtta taaaacaaag aaggtttttg
     1621 ttaagaaatt aatcctcttt cttgataaga aagttgaacc gaaattgcag atactgatat
     1681 atgaaaataa tacccacaat tttgggaata gcgcaagcct caatttaaac aataggtgag
     1741 gacacatgat aatgacctca atgattgtta gaagaaaaga gcctcattac aaaatcgaaa
     1801 aatgaatggt tgggtacaag tttccaaaac atggtaaagt ggactttgcg tatgagacgt
     1861 aaatagaaaa aaacacttgt tatatgtttt ctagaattat tgttgtctct ttatggttgg
     1921 atgatgcaaa atagtaattt cggttagttg ctgtaaaaca ccacgagaca aatagatatg
     1981 gatatttatt aaatcaggaa aaacgtaact ctcggctact ggatggttca gtcacccaac
     2041 gattactggg gagagaaaac agggcaaaag caaagcttaa aggaatccga ttgtcattcg
     2101 gcaatgtgca gcgaaactaa aaaccggata atggacctgt taatcgaaac attgaagata
     2161 aaggaagagg aatcctggca tatcatcaat tgaataagtt gaattaatta tttcaatctc
     2221 attctcactt tctgacttat agtcgctttg ttaaatcata tgtcgactct agaggatccc
     2281 cgggtaaaag gaatgtctcc cttgccagta ctgctagggt ttttctttca aactatggaa
     2341 gcccattcaa gctgcatatt acgattttgt ttttcgcttt tagaaagtgg tttagatgag
     2401 ataatagaaa aattcttgat ctccgacaac gagtactttt attttttttg ctaatcactt
     2461 tactcaatat tagctcgaaa tcgtagaaac gtagacgggt gcgggatacc gagtggtgta
     2521 gttaagaatt tttataaacc acgtggccca aaaatatgaa cccaaaacgt ttatacatga
     2581 gtatacttta agaaggctat accccttcgt gttagatgta gttttagcta cccaacccga
     2641 gtctatgagc ttgacttcag atgtagaagg cattaaatcg ttttgaatat taattaaaaa
     2701 acgatgaaaa ttaaatattt aaaagcaatc atacgctgaa aatttagtgc tgtggctaat
     2761 ccttcaacat ggaaatgcca taaaagtgac tttgacaaaa aaaaaagtat atacaggtag
     2821 taaactcatc tacttcattg actttgttta cagcatgtgg aaggaggaat atttattgct
     2881 aaatcgtagt ttaacattca ataagtaata ctattgaaat tcgacaagat tggccgcatg
     2941 gatgaaaaag aggcattttg ctttgggaga attagttcaa attagaactg aaaaaaaaaa
     3001 ctttacgagg caaaaatgtc ggattgagat cgtaaaagtt cgctcgtcgt cttttgcttt
     3061 gtgattgttt tcatggatac atcttgctgg atatttaaat tttagtacta tgtataagat
     3121 attctataaa tgttttatca cccaaacctg ttagcgcctt cttaattcta ttcaatctgg
     3181 cttttgctct gagactactt cttggacttt cactacttgt tagttatacg gaatttgtgt
     3241 aattagaagt gaaataatcc tttctattag taatgcgagc tgcaattcga gtctaactcc
     3301 ttaaccactc ggacatagtg acttatctga cactcattgt aaattaatat atatataggc
     3361 attttgttta gttaaaggta cttaagtaat tagtataaac gaaccaattt tataatcagg
     3421 aagttaagtg aatggtagca catgtcgtaa aaattgtgaa tttttattga ataatatttt
     3481 aaatacaagc ctttctagac taggtatact cataaacata tatgagcaaa aggatagagg
     3541 agattacatt gcatcttcta caaatttatt tattgccctt tactgaaaaa ttaaataatg
     3601 agtactaaat gataaaaagc gctcagtaca agaaagattg caaaaatatt gcattcttca
     3661 tgaattaaag ttgcatataa ggcatattga aagtaatagt actaaaacag cagttagcga
     3721 aaattaatag aattatattc gcaagacaat tgtgacatta aattaaaaaa ttgtagaatt
     3781 tttactatcc tctttaacgc catgagcctt tataaaaagg ttaaattagt tttaacattc
     3841 tttttttgag taagagttta caatttatca aaacctgtgt tattatattc attagtttca
     3901 atttattagc atctagagaa aaatcaattg gcagttacat tgttggaatt tatgaagaaa
     3961 aagaatctac aacggagaat aagttgctga tcgctttccc taaaattgta tattttgctg
     4021 agcttatttt gacatttcgt tgaagttttc tctaattcgc attcatttta agtaaaacaa
     4081 tgagaaataa aattacaaaa aatacaaatt aaaatacaat ttttagctat aattatagac
     4141 gatgcccttg tatcccattc tgtctcgctt gcccctactt tttatctttt atataccata
     4201 atgaacgctg ccgctactaa ccataccccg attttacatt tcggactccc aaggacgtac
     4261 aaaatagaaa actatagaaa aaaataatca gaaaatagca tgtcatctct ttgtaaaacg
     4321 cgtttgcaag aagaaaggaa acaatggaga agagatcatc catttgtatg taaattttag
     4381 taaacttgaa gaaatcacta acaacttctc ttacttaggg attctatgca aaaccttgta
     4441 aatcatctga tggaggactc gatttaatga attggaaggt tggatttcca ggtggcactt
     4501 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4561 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4621 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4681 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4741 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4801 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4861 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4921 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4981 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5041 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5101 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5161 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5221 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5281 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggatctc
     5341 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5401 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5461 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5521 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5581 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5641 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5701 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5761 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5821 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5881 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5941 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6001 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     6061 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6121 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6181 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6241 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6301 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6361 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
