LOCUS       sequence-224006-        5448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Centromere Plasmid for galactose inducible expression of 
            retron-guide chimeric RNA with flanking HDV-HDV ribozyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5448)
  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 5448)
  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: pZS160 CRISPEY HDV-HDV NotI
FEATURES             Location/Qualifiers
     source          1..5448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..71)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(53..71)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(81..98)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(81..97)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(90..112)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      238..693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(325..344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     535..556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(824..1627)
                     /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(1628..1843)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(1867..1886)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1986..2008
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2046..2064)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2105..2608
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2645..2749
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2750..2818,2819..3610)
                     /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(2968..2987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3781..4369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4270..4289
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacaaataa agaaactgtg acatctgaca catttccacg tacccaattc gccctatagt
       61 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
      121 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
      181 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
      241 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
      301 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
      361 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
      421 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
      481 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
      541 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
      601 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
      661 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
      721 tctgtgcggt atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa
      781 tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc
      841 atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat
      901 cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca
      961 catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac
     1021 cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa
     1081 taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt
     1141 ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt
     1201 cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac
     1261 cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca
     1321 atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact
     1381 tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat
     1441 ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt
     1501 ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat
     1561 taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt
     1621 tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata
     1681 ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt
     1741 gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa
     1801 accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac
     1861 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     1921 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1981 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2041 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2101 gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg
     2161 ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata
     2221 aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc
     2281 aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat
     2341 acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac
     2401 acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg
     2461 tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt
     2521 ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta
     2581 aattataatt atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa
     2641 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2701 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2761 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2821 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2881 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2941 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3001 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3061 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3121 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3181 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3241 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3301 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3361 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3421 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3481 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3541 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3601 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3661 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3721 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3781 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3841 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3901 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3961 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4021 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4081 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4141 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4201 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4261 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4321 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4381 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctggccttt tgctcaagta
     4441 gagggggtaa tttttcccct ttattttgtt catacattct taaattgctt tgcctctcct
     4501 tttggaaagc tatacttcgg agcactgttg agcgaaggct cattagatat attttctgtc
     4561 attttcctta acccaaaaat aagggaaagg gtccaaaaag cgctcggaca actgttgacc
     4621 gtgatccgaa ggactggcta tacagtgttc acaaaatagc caagctgaaa ataatgtgta
     4681 gctatgttca gttagtttgg ctagcaaaga tataaaagca ggtcggaaat atttatgggc
     4741 attattatgc agagcatcaa catgataaaa aaaaacagtt gaatattccc tcaaaagatg
     4801 gccggcatgg tcccagcctc ctcgctggcg ccggctgggc aacaccttcg ggtggcgaat
     4861 gggacttatg cgcaccctta gcgagaggtt tatcattaag gtcaacctct ggatgttgtt
     4921 tcggcatcct gcattgaatc tgagttactg tctgttttcc tgcggccgcg tttcagagct
     4981 atgctggaaa cagcatagca agttgaaata aggctagtcc gttatcaact tgaaaaagtg
     5041 gcaccgagtc ggtgcttttt gatggccggc atggtcccag cctcctcgct ggcgccggct
     5101 gggcaacacc ttcgggtggc gaatgggact taaagaaagt ggaatattca ttcatatcat
     5161 attttttcta ttaactgcct ggtttctttt aaatttttta ttggttgtcg acttgaacgg
     5221 agtgacaata tatatatata tatttaataa tgacatcatt atctgtaaat ctgattctta
     5281 atgctattct agttatgtaa gagtggtcct ttccataaaa aaaaaaaaaa gaaaaaagaa
     5341 ttttaggaat acaatgcagc ttgtaagtaa aatctggaat attcatatcg ccacaacttc
     5401 ttatgcttat aaaagcacta atgcctgaat ttatgttgaa aatatgtg
//
