LOCUS       sequence-198894-        5151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA plasmid to introduce his3 deletion in S. pombe. 
            Linearization with Not1 digestion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5151)
  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 5151)
  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: pYZ149
FEATURES             Location/Qualifiers
     source          1..5151
                     /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(2607..2624)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2607..2623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2616..2638)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2765..3220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2852..2871)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3062..3083
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3246..3350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3351..3419,3420..4211)
                     /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(3569..3588)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4382..4970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4871..4890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5124..5141
                     /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 gcacagtaat atcttatatc tatggaatag aaagaaacgg aaatcgattg
      361 tcatgaaatg atttcccttt ctacgtttcc tttccatttg attataaggt atttacgtta
      421 tagtcaacga aagcttttaa tttccgtcaa ccgatttgaa aggaagaagg atattcgact
      481 tgcaatcgat gcttttagtg cattgcgcga tctttctgct aaccgatttc ctgaatactt
      541 gcttttagta gccggtggct acgatattcg tgtttcagaa aatcgtcgat atttaaaaga
      601 attgcaggaa ttttgtgaac aaaaggatct ttcttatacg actgtaaaag ataattggga
      661 taacataact gttgctcctt ctaccaatgt attgttttta ttatcagttc cttcaaaggt
      721 tcgggatgct ttaatttctt cttctagaat attgctctac actcctgaaa atgaacattt
      781 tggtattgtc cctttggaag ccatgcttcg taaagttcca gtattagccc aaacgaatgg
      841 tgggcccttg gaaacggtta tagatggaaa gaatggttgg cttcgtccta gagacgcgaa
      901 aatttgggga aatgttatat atgaagcgac tactagtact acatatgata ctgctgccat
      961 gggtgaagca gggagtgaat gggtgaagaa tgaattttct acagatgcaa tggctcgaaa
     1021 attcgaatcg gaaataatgt ctggaatacg atcaatcacg cccgaaaaaa gattaatgag
     1081 acgcgtgaat ggactgttgg ctgtctttgt tctatttatg ctattttggg ggacatgcat
     1141 tatcgctgca actgttccct tcgccataat caaattgtat ttcgctcaaa cgtattcaag
     1201 cgtaaaactc ggatttatgc taggaacttg tatagtttca gtgtcatttc taactttcac
     1261 tgtctatgca aagctaacga atctttaatt cttttattac acgggttata atcctttcta
     1321 ttaagattag caaaaaatat ataggcgaga aaagggtaaa aaacaacaaa ggcctgaaat
     1381 cgccagctcc aacaacaaca gtaaacaggg agtacttcat tgctttccaa gcagctaata
     1441 aaatatcttt ctagaagtag ccctcatatc aattattacc caaatataaa gataaaacat
     1501 actcagagaa gtcactatat tctatgttaa tatatatcac gctttctagt ttatcaagtt
     1561 attacatact gtttacagtt cgctgaaatt aaaacacata tttactagac ttttgccgtc
     1621 cgagaaatag atatttgtta cttaaattca taattcatta cattttatga ggttgtttca
     1681 attgctacat gctttttatc tagctatttt attttcacta tacaggctgc atagatgagg
     1741 cagttcaatc cagtcttatt cacaatctat ttcattacct aggataacca ttttggattc
     1801 ataacttttt ggcctacgat ttatgtaaaa gttaaataaa ttaaattgtc gcagcgcttt
     1861 tgctttattt tattcaaaga aaatgcgaat actattgctc aacctttttt aattaaatat
     1921 tgtttaaatc ctttaaacaa aacgatcgga aaatctagcc agtgttccaa gatgaagatg
     1981 caagtttcta ctcgagaatg gatccttaca ttaaggctgg tgctttccaa cccttactac
     2041 acataaggta cttatattcg taacgcattc atccaaacgt gtagctcttt tagattctat
     2101 tagtttcttt tttttagaag atagttgtga atgatttttt tacgcgtttt cactaagaat
     2161 tgttttgctg tacaacctga cttgaaagct taaagatctt gaagacatgg atagtatagg
     2221 ggataatgta acggaaatgg atgcttttgt tcgagatctt actatgaaac cgtatgcaga
     2281 actggagaaa cgaaaagccg aattgcatgc tcaaaaatta aaattggtgc aaaaacgcca
     2341 gcaactgtta cgagacaatt acaatgtatt agtcgattat gctaggaatc aagatgcctt
     2401 ttatcaactt ctcgagaaca gtagacacga ttttaaagag cttgttttac atacaaatca
     2461 gctatatgaa cctgtcaaga gaagccaaaa ctttctaact tcaatttcag agcattacag
     2521 agatgcaaaa ctaatgcatc aagttcagcc gcattaagta gtatactgtt ttagcggccg
     2581 cgggccggcc taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
     2641 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2701 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2761 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2821 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2881 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2941 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3001 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3061 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3121 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3181 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     3241 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3301 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3361 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3421 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3481 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3541 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3601 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3661 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3721 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3781 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3841 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3901 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3961 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4021 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggatct cgcggtatca
     4081 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4141 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4201 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4261 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4321 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4381 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4441 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4501 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4561 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4621 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4681 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4741 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4801 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4861 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4921 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4981 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5041 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5101 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
