LOCUS       sequence-224003-        5424 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 5424)
  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 5424)
  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..5424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(602..1190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(682..701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1361..2152,2153..2221))
                     /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     1984..2003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2222..2326)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2363..2866
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2907..2925
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2963..2985)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3085..3104
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3128..3343
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3344..4147
                     /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"
     rep_origin      complement(4278..4733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4415..4436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4627..4646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4859..4881
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4873..4890
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4874..4890
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4900..4919
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4900..4918
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gctgtttcca gcatagctct gaaacgcggc cgcaggaaaa cagacagtaa ctcagattca
       61 atgcaggatg ccgaaacaac atccagaggt tgaccttaat gataaacctc tcgctaaggg
      121 tgcgcatgac gagctagctc gtttcgtcct cacggactca tcagatgcgc acccttttga
      181 gggaatattc aactgttttt ttttatcatg ttgatgctct gcataataat gcccataaat
      241 atttccgacc tgcttttata tctttgctag ccaaactaac tgaacatagc tacacattat
      301 tttcagcttg gctattttgt gaacactgta tagccagtcc ttcggatcac ggtcaacagt
      361 tgtccgagcg ctttttggac cctttccctt atttttgggt taaggaaaat gacagaaaat
      421 atatctaatg agccttcgct caacagtgct ccgaagtata gctttccaaa aggagaggca
      481 aagcaattta agaatgtatg aacaaaataa aggggaaaaa ttaccccctc tacttgagca
      541 aaaggccagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcct
      601 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      661 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      721 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      781 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      841 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      901 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      961 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1021 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1081 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1141 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1201 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1261 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1321 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1381 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1441 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1501 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1561 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1621 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1681 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1741 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1801 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1861 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1921 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1981 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2041 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2101 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2161 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2221 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2281 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2341 aagtgccacc tgggtccttt tcatcacgtg ctataaaaat aattataatt taaatttttt
     2401 aatataaata tataaattaa aaatagaaag taaaaaaaga aattaaagaa aaaatagttt
     2461 ttgttttccg aagatgtaaa agactctagg gggatcgcca acaaatacta ccttttatct
     2521 tgctcttcct gctctcaggt attaatgccg aattgtttca tcttgtctgt gtagaagacc
     2581 acacacgaaa atcctgtgat tttacatttt acttatcgtt aatcgaatgt atatctattt
     2641 aatctgcttt tcttgtctaa taaatatata tgtaaagtac gctttttgtt gaaatttttt
     2701 aaacctttgt ttattttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt
     2761 ctaaaatcca aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc
     2821 catcattaaa agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc taagaaacca
     2881 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     2941 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3001 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3061 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     3121 ccaccttttc aattcatcat ttttttttta ttcttttttt tgatttcggt ttccttgaaa
     3181 tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag
     3241 attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac
     3301 ccaactgcac agaacaaaaa cctgcaggaa acgaagataa atcatgtcga aagctacata
     3361 taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca
     3421 cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga
     3481 gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac
     3541 tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt
     3601 tttactcttc gaagacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc
     3661 tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg
     3721 cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg
     3781 ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa
     3841 gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag
     3901 agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt
     3961 agatgacaag ggagacgcat tgggtcaaca gtatagaacc gtggatgatg tggtctctac
     4021 aggatctgac attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt
     4081 agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca
     4141 aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc
     4201 aatttaatta tatcagttat taccctatgc ggtgtgaaat accgcacaga tgcgtaagga
     4261 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4321 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4381 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4441 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4501 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4561 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4621 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4681 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     4741 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4801 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     4861 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
     4921 gaattgggta cgtggaaatg tgtcagatgt cacagtttct ttatttgtga cacatatttt
     4981 caacataaat tcaggcatta gtgcttttat aagcataaga agttgtggcg atatgaatat
     5041 tccagatttt acttacaagc tgcattgtat tcctaaaatt cttttttctt tttttttttt
     5101 ttatggaaag gaccactctt acataactag aatagcatta agaatcagat ttacagataa
     5161 tgatgtcatt attaaatata tatatatata ttgtcactcc gttcaagtcg acaaccaata
     5221 aaaaatttaa aagaaaccag gcagttaata gaaaaaatat gatatgaatg aatattccac
     5281 tttctttaag tcccattcgc cacccgaagg tgttgcccag ccggcgccag cgaggaggct
     5341 gggaccatgc cggccatcaa aaagcaccga ctcggtgcca ctttttcaag ttgataacgg
     5401 actagcctta tttcaacttg ctat
//
