LOCUS       sequence-198900-        4897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA plasmid to introduce lys9 deletion in S. pombe. 
            Linearization with Not1 digestion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4897)
  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 4897)
  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: pYZ172
FEATURES             Location/Qualifiers
     source          1..4897
                     /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(2353..2370)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2353..2369)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2362..2384)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2511..2966
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2598..2617)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2808..2829
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2992..3096
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3097..3165,3166..3957)
                     /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(3315..3334)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4128..4716
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4617..4636
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4870..4887
                     /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 gcacagtgcg agcacttgga ttttaacctc ctaatttgta aacggtactg
      361 tatctttctt tttataattt ttgagtcgaa gcatgttctg ctgcataaca cattcatatt
      421 tgtcaattct acttaaatag attttaaatc attgcattcg aagataaaat tatggtagct
      481 tagtgttcct acggatctta atgtatttat atgttcactt tataatacga tacaaaacaa
      541 attttatctc tcctttgttt caaattgact ggcaaaaaaa aataacaaat aactaatcac
      601 aaaaacctaa ggaatatgaa aaatagttgc taattttcgt agaaacttgg atcttgatta
      661 gaaatgaacc aaattatgca tcattcaaat agattgtagt gtgtatggtg cagcgggaac
      721 gttataatta accatcacct atagtctcaa aatgttacgc aaattgccca tcaattttgc
      781 gaaatggact gtaaaaaagg tgccagttca gcagaaaaga tttaatagtc aacaaaaaga
      841 aatctcacct catgtaagct tttctaaata cgctttaggt cataaatttt atctgtgcta
      901 aaattcattc aagcattttt gcattgtttt atttaacatt attttattaa cgatcataaa
      961 gattatgttt tataaaaact acgctagacc tcttggtaaa gtgacgcttt ttgccttagc
     1021 aacttattat ggattggaaa tagtttggtg gaaattagat gcatcagaac aggaagctat
     1081 taaaaacagt aagttgttga tatgtgaaag ctcttttagt ttgttgacat ttaggcgaat
     1141 tacggaattt agagaatgcg aaataaaaac acgagatttg tatgacccag aaatttagta
     1201 acgtatatgg cttaaagaag cgagattacg atttgttaat agttcagtat gtcaaaaatt
     1261 ctttgaatag ctaaatgcta cagacgaaac actcagtcaa aaggtcctcg agttgttgaa
     1321 tggtggtttt ttatgcagtc gggaatgaaa gcagctgtaa tttccttgca taatgagcct
     1381 aaataccttt ttacatagaa gttaaaattt atgagatttt aaattaaaaa atcatcaaaa
     1441 gtgcaaattt tcaagtacat atacgaataa atttaattat aaagtaaacc taaacgttgt
     1501 ttaaagctaa ttgcttcttg gtaaccgatt gcaacatgcc atcatttatt tacattgaca
     1561 acccgtaaat atccataata ggtagaatgt tatcgacaat gaatttcaaa tgcatatata
     1621 tgtttaagta aaacaaaata ctctacacta gcagtcccgc aatgggaaaa taaagaggca
     1681 gcaaatatga aaagcttccg tcatttcttt ttcggtcttg ctaaaaaaat aaaaccacac
     1741 atagcttaaa aaaagtaaca atgactagac atagcatgtc atgaaaatca actgtttctt
     1801 taagcttttg tagataaacc gaaaattgac aaattgaaag ttccacctaa aaatgcatca
     1861 ctgttggatc ctcccaaata atttaataga aattatttaa attgcatgtt ctaaatcaac
     1921 aggtattaaa agatattact ggggttgttg cgaatagcga gaaagagggt tttgttgctg
     1981 ggaaaactgc tgatgcattg ggtttccctg acgcccttgc tgagtttgat acatcatagc
     2041 gtttggctgt gaataaagtc tctgttgctg atcgaattgc cgaggagatc caggtcttct
     2101 tgacatacca tatgcagaag gatcaaacat ttgagactgt tgcatattag gtcgcatatt
     2161 accttgaaat tggggcattt gatatcccat aggggatgac tgcaatcgga aagctgagga
     2221 tccagattcg agatgcgtag gtggataacg gtataaatgc tcagccatgt acggtgctac
     2281 catgaccggt ctttgcatat tcatctggtt aacattttag cggccgcggg ccggcctacc
     2341 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2401 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2461 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     2521 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2581 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2641 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2701 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2761 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2821 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2881 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2941 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     3001 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3061 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3121 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3181 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3241 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3301 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3361 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3421 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3481 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3541 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3601 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3661 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3721 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3781 attgctgata aatctggagc cggtgagcgt ggatctcgcg gtatcattgc agcactgggg
     3841 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3901 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3961 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4021 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4081 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4141 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4201 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4261 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4321 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4381 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4441 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4501 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4561 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4621 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4681 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4741 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4801 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4861 accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
