LOCUS       sequence-216021-        5428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Centromere Plasmid for galactose inducible expression of 
            retron-guide chimeric RNA with flanking HH-HDV ribozyme .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5428)
  AUTHORS   Sharon E, Chen SA, Khosla NM, Smith JD, Pritchard JK, Fraser HB
  TITLE     Functional Genetic Variants Revealed by Massively Parallel Precise 
            Genome Editing.
  JOURNAL   Cell. 2018 Sep 18. pii: S0092-8674(18)31118-8. doi: 
            10.1016/j.cell.2018.08.057.
  PUBMED    30245013
REFERENCE   2  (bases 1 to 5428)
  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: "Cell. 2018 
            Sep 18. pii: S0092-8674(18)31118-8. doi: 
            10.1016/j.cell.2018.08.057."
COMMENT     Sequence Label: pZS165 CRISPEY HH-HDV NotI
FEATURES             Location/Qualifiers
     source          1..5428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        127..231
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(232..300,301..1092)
                     /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(450..469)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1263..1851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1752..1771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2950..2969)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2951..2969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2979..2996)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2979..2995)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2988..3010)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3140..3595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3227..3246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3437..3458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3726..4529)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4530..4750)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4776..4795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4895..4917
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4955..4973)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5015..5428,1..90)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ttttctttaa tttctttttt tactttctat ttttaattta tatatttata ttaaaaaatt
       61 taaattataa ttatttttat agcacgtgat gaaaaggacc caggtggcac ttttcgggga
      121 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      181 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      241 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      301 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
      361 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
      421 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
      481 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
      541 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
      601 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
      661 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
      721 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
      781 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
      841 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
      901 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      961 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     1021 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1081 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     1141 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1201 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1261 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1321 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1381 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     1441 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1501 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1561 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1621 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1681 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1741 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1801 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1861 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcaagt agagggggta
     1921 atttttcccc tttattttgt tcatacattc ttaaattgct ttgcctctcc ttttggaaag
     1981 ctatacttcg gagcactgtt gagcgaaggc tcattagata tattttctgt cattttcctt
     2041 aacccaaaaa taagggaaag ggtccaaaaa gcgctcggac aactgttgac cgtgatccga
     2101 aggactggct atacagtgtt cacaaaatag ccaagctgaa aataatgtgt agctatgttc
     2161 agttagtttg gctagcaaag atataaaagc aggtcggaaa tatttatggg cattattatg
     2221 cagagcatca acatgataaa aaaaaacagt tgaatattcc ctcaaaaggg tgcgcatctg
     2281 atgagtccgt gaggacgaaa cgagctagct cgtcatgcgc acccttagcg agaggtttat
     2341 cattaaggtc aacctctgga tgttgtttcg gcatcctgca ttgaatctga gttactgtct
     2401 gttttcctgc ggccgcgttt cagagctatg ctggaaacag catagcaagt tgaaataagg
     2461 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttgat ggccggcatg
     2521 gtcccagcct cctcgctggc gccggctggg caacaccttc gggtggcgaa tgggacttaa
     2581 agaaagtgga atattcattc atatcatatt ttttctatta actgcctggt ttcttttaaa
     2641 ttttttattg gttgtcgact tgaacggagt gacaatatat atatatatat atttaataat
     2701 gacatcatta tctgtaaatc tgattcttaa tgctattcta gttatgtaag agtggtcctt
     2761 tccataaaaa aaaaaaaaaa gaaaaaagaa ttttaggaat acaatgcagc ttgtaagtaa
     2821 aatctggaat attcatatcg ccacaacttc ttatgcttat aaaagcacta atgcctgaat
     2881 ttatgttgaa aatatgtgtc acaaataaag aaactgtgac atctgacaca tttccacgta
     2941 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     3001 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3061 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3121 ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3181 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3241 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3301 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3361 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3421 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3481 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3541 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcctga
     3601 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg taataactga
     3661 tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca
     3721 gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa
     3781 cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata
     3841 atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct
     3901 cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt
     3961 ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg
     4021 tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct
     4081 aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta
     4141 acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg
     4201 tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct
     4261 tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc
     4321 atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat
     4381 gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt
     4441 gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca
     4501 gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt
     4561 ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt
     4621 atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc
     4681 gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa
     4741 ttgaattgaa aagctgtggt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4801 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4861 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4921 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4981 gttaatgtca tgataataat ggtttcttag gacggatcgc ttgcctgtaa cttacacgcg
     5041 cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat
     5101 gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa
     5161 aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta
     5221 ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa
     5281 atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat
     5341 acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct
     5401 tttacatctt cggaaaacaa aaactatt
//
