LOCUS       sequence-198862-        5015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA plasmid to introduce leu1 deletion in S. pombe. 
            Linearization with Not1 digestion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5015)
  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 5015)
  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: pYZ145
FEATURES             Location/Qualifiers
     source          1..5015
                     /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"
     primer_bind     240..259
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        240..258
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2471..2488)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2471..2487)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2480..2502)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2629..3084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2716..2735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2926..2947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3110..3214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3215..3283,3284..4075)
                     /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(3433..3452)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4246..4834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4735..4754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4988..5005
                     /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 gccaagctct
      241 aatacgactc actataggga aagcttgcat gcctgcaggt cgactctaga ggatcggccg
      301 gcccgcggcc gcacagttta atggttgtat agcgtacaca tttgtaaata tagataagga
      361 agccttggga ggacgttatg tcaacatctc acaagtccgc attgtttaca tttagcttat
      421 ggcgcaaatt cgacaagaag cattgaaaaa acaaaagtct gagacgcaaa aatcaacaga
      481 agggttcgtt gatattggaa atttggctca atggacctgc tcgagcgaaa aatccggatt
      541 tcctatacgc ttggtgcggg atgataatat cgatacatat tggcagtaag tgtatttgta
      601 taaatatagt cagggaagaa aatgttttac taactataat tttctttgtc tttaaggtct
      661 gacggctctc aacctcatac catccatatt aaatttgtaa agcgagtatc gattaagtat
      721 gtgagtatgt accttcaata cacccttgat gaatcttata ctcccagtac tttgcgaata
      781 tcagctggga cagggtttca agatttggaa attgttacaa cagttcaagt cgaggaacca
      841 actggttggg tacatgtgcc agttggtgat tttgggcgaa atggcttgtt agatgtgcat
      901 ttaatacaaa tcaaaatact tgcgaaccat caaagtggaa aggatagtca tgttcgtttg
      961 ataaaaatat atgcgccaga aatgtacgta ttttgcatag caaaatcttc attaatataa
     1021 aatgtgctaa cactgatgca gagagcagcc cgcgatagct gtggatgaaa ttccctacac
     1081 aagtttacaa ttcataagta gaaaccaatt acggtaatgc aaaagtcatg tctctggctc
     1141 tctcaaagaa tcgggttgtc atcgttacaa taataattgt tacgttctcg aatttttatg
     1201 gtttttttct tcttttcccc tattatttat gttacaggtt acttcgttta aaaagaaata
     1261 attaatgata ttatgtttga aacttttcat agtaaaaaaa gagtaagtac acagcgacgc
     1321 gtagcatgtt gtaaaacttt agaaaaatac ttgtagaatg agatgcgttg tagtaacata
     1381 ataattgacg atttatttat tttcgcaact tttactatgc tttctaaata tgggtttaat
     1441 tggaactcgt atcctaccat ttacaatgtt catccaattt tttcagattg tactgtaaat
     1501 agcgtttgaa aacaccaaat tttagaagct aatcactctc atcataatcg tctacatcct
     1561 catcgttatc gacgataaaa gaatcatctt gcatgctggg ttcatccatg ctatcaaacg
     1621 agggatcaac gtaaataggt gttttcactg tagccgctgc tcttctggtt ggcctctttc
     1681 taatcggaga atctgaatct tctggtggct ctgcgttagt cgaactagct tttggagttg
     1741 aactactacc tggaataata aaatcatcat cgtcatcttc aggtgattgt ttctttaccg
     1801 agcttgcttt tttcccttta ttcttcgcag aagccttcgt ggatgttatg gtggaaggtt
     1861 tcaaactgct aggcaacaaa tcatcttcat cgtctgaaga aaatatggta gtagcaactg
     1921 gtttattagt ctttcttcct cttccagacg ccgaggctgc tatttttttg acgggttttt
     1981 tactacctgc gtcttcagag tcaacagatt gacttctttt tcttgatttt ccactatcac
     2041 tgctatccaa tcccgggctc ttagatatgc gattttcttc aactgataag ccatgagagt
     2101 tatcctctgt cttgacaatg tttatgtcag atgatttctc aggttctttc gacgctgcga
     2161 actcaagtaa agtttgttgc tttcgatttg ttgtagatgg tttggattcg ctgctagctt
     2221 cttttttaac agcagtactt gaggaggatc cggcaatagc cctgggtttc ctagtaccag
     2281 tggatttacc tcgaggcttc tttttcgttc gatttacaaa atctctagag gattgctctt
     2341 cttctaacat ttctctctga atatcatcca taaccttatt ccaagcatgc tcaaatgcat
     2401 ccaaatcatg aagccacaat tctttaggag ttttttagcg gccgcgggcc ggcctaccga
     2461 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     2521 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     2581 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     2641 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2701 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2761 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2821 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2881 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2941 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3001 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3061 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3181 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3241 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3301 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3361 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3421 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3481 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3541 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3601 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3661 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3721 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3781 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3841 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3901 tgctgataaa tctggagccg gtgagcgtgg atctcgcggt atcattgcag cactggggcc
     3961 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4021 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4081 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4141 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4201 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4261 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4321 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4381 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4441 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4501 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4561 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4621 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4681 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4741 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4801 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4861 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4921 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4981 cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
