LOCUS       sequence-216022-        5452 bp    DNA     circular SYN 14-APR-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 5452)
  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 5452)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..5452
                     /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(2974..2993)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2975..2993)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3003..3020)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3003..3019)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3012..3034)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3164..3619
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3251..3270)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3461..3482
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3750..4553)
                     /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(4554..4774)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4800..4819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4919..4941
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4979..4997)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5039..5452,1..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 ctcaaaagat ggccggcatg
     2281 gtcccagcct cctcgctggc gccggctggg caacaccttc gggtggcgaa tgggacttat
     2341 gcgcaccctt agcgagaggt ttatcattaa ggtcaacctc tggatgttgt ttcggcatcc
     2401 tgcattgaat ctgagttact gtctgttttc ctgcggccgc gtttcagagc tatgctggaa
     2461 acagcatagc aagttgaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     2521 cggtgctttt tgatggccgg catggtccca gcctcctcgc tggcgccggc tgggcaacac
     2581 cttcgggtgg cgaatgggac ttaaagaaag tggaatattc attcatatca tattttttct
     2641 attaactgcc tggtttcttt taaatttttt attggttgtc gacttgaacg gagtgacaat
     2701 atatatatat atatatttaa taatgacatc attatctgta aatctgattc ttaatgctat
     2761 tctagttatg taagagtggt cctttccata aaaaaaaaaa aaaagaaaaa agaattttag
     2821 gaatacaatg cagcttgtaa gtaaaatctg gaatattcat atcgccacaa cttcttatgc
     2881 ttataaaagc actaatgcct gaatttatgt tgaaaatatg tgtcacaaat aaagaaactg
     2941 tgacatctga cacatttcca cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     3001 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3061 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3121 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc
     3181 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3241 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3301 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3361 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3421 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3481 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3541 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3601 atattaacgt ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3661 cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     3721 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     3781 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     3841 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     3901 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     3961 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     4021 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     4081 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     4141 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     4201 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     4261 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     4321 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     4381 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     4441 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     4501 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     4561 cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     4621 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     4681 tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg aaatcaaaaa
     4741 aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg tggtatggtg cactctcagt
     4801 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4861 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4921 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4981 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttaggacgga
     5041 tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt
     5101 tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag
     5161 gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa
     5221 aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag atatacattc
     5281 gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac
     5341 aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat
     5401 ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta tt
//
