LOCUS       pSAC212_for_addgene     5992 bp ds-DNA     circular     16-JUN-2022
DEFINITION  .
KEYWORDS    "accession:U03450" "marker:AmpR" "organism:Saccharomyces           
            cerevisiae" "ref:pmid\:2659436"
FEATURES             Location/Qualifiers
     CDS             complement(242..1045)
                     /label="URA3"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     CDS             complement(242..1045)
                     /label="Translation 1045-242"
     promoter        complement(1046..1266)
                     /label="URA3 promoter"
                     /ApEinfo_revcolor=#ffffff
                     /ApEinfo_fwdcolor=#ffffff
     misc_feature    1531..2034
                     /label="CEN/ARS"
                     /ApEinfo_revcolor=#454f59
                     /ApEinfo_fwdcolor=#454f59
     promoter        2071..2175
                     /label="AmpR promoter"
                     /ApEinfo_revcolor=#ffffff
                     /ApEinfo_fwdcolor=#ffffff
     CDS             2176..2244
                     /label="AmpR"
                     /ApEinfo_revcolor=#c4df9b
                     /ApEinfo_fwdcolor=#c4df9b
     CDS             2245..3036
                     /label="AmpR"
                     /ApEinfo_revcolor=#c4df9b
                     /ApEinfo_fwdcolor=#c4df9b
     rep_origin      3207..3795
                     /label="ori"
                     /ApEinfo_revcolor=#fff79a
                     /ApEinfo_fwdcolor=#fff79a
     promoter        3851..4211
                     /label="pGal7"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    3930..3939
                     /label="enhancer"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    4173..4211
                     /label="potential 5 UTR'"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    4212..4278
                     /label="drz-CIV1"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    4279..4360
                     /label="msr"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    4280..4291
                     /label="inverted repeat"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    4373..4409
                     /label="5' homology for barcoding guide"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    4402..4417
                     /label="GA 5' homology"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    4404..4943
                     /label="Twist Synthesis"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    4410..4417
                     /label="SfiI.A"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    4418..4427
                     /label="place holder 10 bp PBC"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    4428..4433
                     /label="SphI"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    4428..4795
                     /label="sgRNA-HDV-msr insert"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    complement(4434..4458)
                     /label="oligo amplification R"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    4434..4463
                     /label="GA 3' homology"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    4434..4477
                     /label="3' homology YBR209W"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    4478..4511
                     /label="constant synthesis"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    4491..4491
                     /label="Mutated base avoid terminator"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    complement(4498..4498)
                     /label="RT start"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    complement(4500..4511)
                     /label="inverted repeat"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    4512..4531
                     /label="YBR209W_guide3"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    4532..4622
                     /label="sgRNA"
                     /ApEinfo_revcolor=#7ea7d8
                     /ApEinfo_fwdcolor=#7ea7d8
     misc_feature    4536..4536
                     /label="T-C"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    4567..4567
                     /label="A-G"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    4623..4693
                     /label="HDV-Ribozyme"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    4694..4775
                     /label="msr"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    4695..4706
                     /label="inverted repeat"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    4788..4795
                     /label="AscI"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    4796..4875
                     /label="donor DNA for ade2 KO"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     CDS             4796..4875
                     /label="Translation 4796-4875"
     misc_feature    4876..4909
                     /label="constant synthesis"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    4889..4889
                     /label="Mutated base avoid terminator"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    complement(4896..4896)
                     /label="RT start"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    complement(4898..4909)
                     /label="inverted repeat"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    4910..4927
                     /label="guide for targeting ade2"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    4928..4946
                     /label="GA 5' homology"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    4928..5018
                     /label="sgRNA"
                     /ApEinfo_revcolor=#7ea7d8
                     /ApEinfo_fwdcolor=#7ea7d8
     misc_feature    4932..4932
                     /label="T-C"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    4963..4963
                     /label="A-G"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    5019..5086
                     /label="drz-Spur3"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     terminator      5087..5445
                     /label="tGal7"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     promoter        complement(5459..5477)
                     /label="T7_promoter"
                     /ApEinfo_revcolor=#8493ca
                     /ApEinfo_fwdcolor=#8493ca
     misc_feature    complement(5487..5503)
                     /label="M13_forward20_primer"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     CDS             5504..5504
                     /label="lacZα-Broken"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     CDS             5505..5865
                     /label="lacZα"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     rep_origin      5648..111
                     /label="f1 ori"
                     /ApEinfo_revcolor=#fff79a
                     /ApEinfo_fwdcolor=#fff79a
ORIGIN
        1 ATTCTTTTGA TTTATAAGGG ATTTTGCCGA TTTCGGCCTA TTGGTTAAAA AATGAGCTGA
       61 TTTAACAAAA ATTTAACGCG AATTTTAACA AAATATTAAC GTTTACAATT TCCTGATGCG
      121 GTATTTTCTC CTTACGCATC TGTGCGGTAT TTCACACCGC ATAGGGTAAT AACTGATATA
      181 ATTAAATTGA AGCTCTAATT TGTGAGTTTA GTATACATGC ATTTACTTAT AATACAGTTT
      241 TTTAGTTTTG CTGGCCGCAT CTTCTCAAAT ATGCTTCCCA GCCTGCTTTT CTGTAACGTT
      301 CACCCTCTAC CTTAGCATCC CTTCCCTTTG CAAATAGTCC TCTTCCAACA ATAATAATGT
      361 CAGATCCTGT AGAGACCACA TCATCCACGG TTCTATACTG TTGACCCAAT GCGTCTCCCT
      421 TGTCATCTAA ACCCACACCG GGTGTCATAA TCAACCAATC GTAACCTTCA TCTCTTCCAC
      481 CCATGTCTCT TTGAGCAATA AAGCCGATAA CAAAATCTTT GTCGCTCTTC GCAATGTCAA
      541 CAGTACCCTT AGTATATTCT CCAGTAGATA GGGAGCCCTT GCATGACAAT TCTGCTAACA
      601 TCAAAAGGCC TCTAGGTTCC TTTGTTACTT CTTCTGCCGC CTGCTTCAAA CCGCTAACAA
      661 TACCTGGGCC CACCACACCG TGTGCATTCG TAATGTCTGC CCATTCTGCT ATTCTGTATA
      721 CACCCGCAGA GTACTGCAAT TTGACTGTAT TACCAATGTC AGCAAATTTT CTGTCTTCGA
      781 AGAGTAAAAA ATTGTACTTG GCGGATAATG CCTTTAGCGG CTTAACTGTG CCCTCCATGG
      841 AAAAATCAGT CAAGATATCC ACATGTGTTT TTAGTAAACA AATTTTGGGA CCTAATGCTT
      901 CAACTAACTC CAGTAATTCC TTGGTGGTAC GAACATCCAA TGAAGCACAC AAGTTTGTTT
      961 GCTTTTCGTG CATGATATTA AATAGCTTGG CAGCAACAGG ACTAGGATGA GTAGCAGCAC
     1021 GTTCCTTATA TGTAGCTTTC GACATGATTT ATCTTCGTTT CCTGCAGGTT TTTGTTCTGT
     1081 GCAGTTGGGT TAAGAATACT GGGCAATTTC ATGTTTCTTC AACACTACAT ATGCGTATAT
     1141 ATACCAATCT AAGTCTGTGC TCCTTCCTTC GTTCTTCCTT CTGTTCGGAG ATTACCGAAT
     1201 CAAAAAAATT TCAAAGAAAC CGAAATCAAA AAAAAGAATA AAAAAAAAAT GATGAATTGA
     1261 ATTGAAAAGC TGTGGTATGG TGCACTCTCA GTACAATCTG CTCTGATGCC GCATAGTTAA
     1321 GCCAGCCCCG ACACCCGCCA ACACCCGCTG ACGCGCCCTG ACGGGCTTGT CTGCTCCCGG
     1381 CATCCGCTTA CAGACAAGCT GTGACCGTCT CCGGGAGCTG CATGTGTCAG AGGTTTTCAC
     1441 CGTCATCACC GAAACGCGCG AGACGAAAGG GCCTCGTGAT ACGCCTATTT TTATAGGTTA
     1501 ATGTCATGAT AATAATGGTT TCTTAGGACG GATCGCTTGC CTGTAACTTA CACGCGCCTC
     1561 GTATCTTTTA ATGATGGAAT AATTTGGGAA TTTACTCTGT GTTTATTTAT TTTTATGTTT
     1621 TGTATTTGGA TTTTAGAAAG TAAATAAAGA AGGTAGAAGA GTTACGGAAT GAAGAAAAAA
     1681 AAATAAACAA AGGTTTAAAA AATTTCAACA AAAAGCGTAC TTTACATATA TATTTATTAG
     1741 ACAAGAAAAG CAGATTAAAT AGATATACAT TCGATTAACG ATAAGTAAAA TGTAAAATCA
     1801 CAGGATTTTC GTGTGTGGTC TTCTACACAG ACAAGATGAA ACAATTCGGC ATTAATACCT
     1861 GAGAGCAGGA AGAGCAAGAT AAAAGGTAGT ATTTGTTGGC GATCCCCCTA GAGTCTTTTA
     1921 CATCTTCGGA AAACAAAAAC TATTTTTTCT TTAATTTCTT TTTTTACTTT CTATTTTTAA
     1981 TTTATATATT TATATTAAAA AATTTAAATT ATAATTATTT TTATAGCACG TGATGAAAAG
     2041 GACCCAGGTG GCACTTTTCG GGGAAATGTG CGCGGAACCC CTATTTGTTT ATTTTTCTAA
     2101 ATACATTCAA ATATGTATCC GCTCATGAGA CAATAACCCT GATAAATGCT TCAATAATAT
     2161 TGAAAAAGGA AGAGTATGAG TATTCAACAT TTCCGTGTCG CCCTTATTCC CTTTTTTGCG
     2221 GCATTTTGCC TTCCTGTTTT TGCTCACCCA GAAACGCTGG TGAAAGTAAA AGATGCTGAA
     2281 GATCAGTTGG GTGCACGAGT GGGTTACATC GAACTGGATC TCAACAGCGG TAAGATCCTT
     2341 GAGAGTTTTC GCCCCGAAGA ACGTTTTCCA ATGATGAGCA CTTTTAAAGT TCTGCTATGT
     2401 GGCGCGGTAT TATCCCGTAT TGACGCCGGG CAAGAGCAAC TCGGTCGCCG CATACACTAT
     2461 TCTCAGAATG ACTTGGTTGA GTACTCACCA GTCACAGAAA AGCATCTTAC GGATGGCATG
     2521 ACAGTAAGAG AATTATGCAG TGCTGCCATA ACCATGAGTG ATAACACTGC GGCCAACTTA
     2581 CTTCTGACAA CGATCGGAGG ACCGAAGGAG CTAACCGCTT TTTTGCACAA CATGGGGGAT
     2641 CATGTAACTC GCCTTGATCG TTGGGAACCG GAGCTGAATG AAGCCATACC AAACGACGAG
     2701 CGTGACACCA CGATGCCTGT AGCAATGGCA ACAACGTTGC GCAAACTATT AACTGGCGAA
     2761 CTACTTACTC TAGCTTCCCG GCAACAATTA ATAGACTGGA TGGAGGCGGA TAAAGTTGCA
     2821 GGACCACTTC TGCGCTCGGC CCTTCCGGCT GGCTGGTTTA TTGCTGATAA ATCTGGAGCC
     2881 GGTGAGCGTG GGTCTCGCGG TATCATTGCA GCACTGGGGC CAGATGGTAA GCCCTCCCGT
     2941 ATCGTAGTTA TCTACACGAC GGGGAGTCAG GCAACTATGG ATGAACGAAA TAGACAGATC
     3001 GCTGAGATAG GTGCCTCACT GATTAAGCAT TGGTAACTGT CAGACCAAGT TTACTCATAT
     3061 ATACTTTAGA TTGATTTAAA ACTTCATTTT TAATTTAAAA GGATCTAGGT GAAGATCCTT
     3121 TTTGATAATC TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC
     3181 CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC
     3241 TTGCAAACAA AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA
     3301 ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTCCTTCTA
     3361 GTGTAGCCGT AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT
     3421 CTGCTAATCC TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG
     3481 GACTCAAGAC GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC
     3541 ACACAGCCCA GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA
     3601 TGAGAAAGCG CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG
     3661 GTCGGAACAG GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT
     3721 CCTGTCGGGT TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG
     3781 CGGAGCCTAT GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG
     3841 CCTTTTGCTC AAGTAGAGGG GGTAATTTTT CCCCTTTATT TTGTTCATAC ATTCTTAAAT
     3901 TGCTTTGCCT CTCCTTTTGG AAAGCTATAC TTCGGAGCAC TGTTGAGCGA AGGCTCATTA
     3961 GATATATTTT CTGTCATTTT CCTTAACCCA AAAATAAGGG AAAGGGTCCA AAAAGCGCTC
     4021 GGACAACTGT TGACCGTGAT CCGAAGGACT GGCTATACAG TGTTCACAAA ATAGCCAAGC
     4081 TGAAAATAAT GTGTAGCTAT GTTCAGTTAG TTTGGCTAGC AAAGATATAA AAGCAGGTCG
     4141 GAAATATTTA TGGGCATTAT TATGCAGAGC ATCAACATGA TAAAAAAAAA CAGTTGAATA
     4201 TTCCCTCAAA AGAGGTGCTT GTAGATAACC TCCACGATGG TGCACCTTGG GCAACACAAA
     4261 AGTGGCAAAT CATCTACAAT GCGCACCCTT AGCGAGAGGT TTATCATTAA GGTCAACCTC
     4321 TGGATGTTGT TTCGGCATCC TGCATTGAAT CTGAGTTACT GTCTGTTTga acTGCAGCCA
     4381 AAGATGCGTG CCGTTGCAGT TAGCTAACAg gccATGCTAG CATCGATgca tgcACCTGGC
     4441 GTTCGGCGAT CGCCATAAGA GATCTGCCAA TTTTAAAttt AAACCCGTTT CTTCTGACGT
     4501 AAGGGTGCGC AGTTGCAGTT AGCTAACAAC CgtttCagag ctaTGCTGga aaCAGCAtag
     4561 caagttGaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     4621 ttGATGGCCG GCATGGTCCC AGCCTCCTCG CTGGCGCCGG CTGGGCAACA CCTTCGGGTG
     4681 GCGAATGGGA CTTATGCGCA CCCTTAGCGA GAGGTTTATC ATTAAGGTCA ACCTCTGGAT
     4741 GTTGTTTCGG CATCCTGCAT TGAATCTGAG TTACTGTCTG TTTTCCTggc gcgccACAGG
     4801 GGAATTGCTT ATTAACGAAA TTGCCTGAAG GCCTCACAAC TCTGGACATT ATACCATTGA
     4861 TGCTTGCGTC ACTTCAGGAA ACCCGTTTCT TCTGACGTAA GGGTGCGCAA TTAACGAAAT
     4921 TGCCCCAgtt tCagagctaT GCTGgaaaCA GCAtagcaag ttGaaataag gctagtccgt
     4981 tatcaacttg aaaaagtggc accgagtcgg tgctttttGA CACTGAGTGA GAAACGTCCC
     5041 CGTCGTAGTG TCGGTAATGC GTTGTTTCAA CGTAGCCAAT TCTCACAAAG AAAGTGGAAT
     5101 ATTCATTCAT ATCATATTTT TTCTATTAAC TGCCTGGTTT CTTTTAAATT TTTTATTGGT
     5161 TGTCGACTTG AACGGAGTGA CAATATATAT ATATATATAT TTAATAATGA CATCATTATC
     5221 TGTAAATCTG ATTCTTAATG CTATTCTAGT TATGTAAGAG TGGTCCTTTC CATAAAAAAA
     5281 AAAAAAAAGA AAAAAGAATT TTAGGAATAC AATGCAGCTT GTAAGTAAAA TCTGGAATAT
     5341 TCATATCGCC ACAACTTCTT ATGCTTATAA AAGCACTAAT GCCTGAATTT ATGTTGAAAA
     5401 TATGTGTCAC AAATAAAGAA ACTGTGACAT CTGACACATT TCCACgtacc caattcgccc
     5461 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacAACGTCG TGACTGGGAA
     5521 AACCCTGGCG TTACCCAACT TAATCGCCTT GCAGCACATC CCCCTTTCGC CAGCTGGCGT
     5581 AATAGCGAAG AGGCCCGCAC CGATCGCCCT TCCCAACAGT TGCGCAGCCT GAATGGCGAA
     5641 TGGCGCGACG CGCCCTGTAG CGGCGCATTA AGCGCGGCGG GTGTGGTGGT TACGCGCAGC
     5701 GTGACCGCTA CACTTGCCAG CGCCCTAGCG CCCGCTCCTT TCGCTTTCTT CCCTTCCTTT
     5761 CTCGCCACGT TCGCCGGCTT TCCCCGTCAA GCTCTAAATC GGGGGCTCCC TTTAGGGTTC
     5821 CGATTTAGTG CTTTACGGCA CCTCGACCCC AAAAAACTTG ATTAGGGTGA TGGTTCACGT
     5881 AGTGGGCCAT CGCCCTGATA GACGGTTTTT CGCCCTTTGA CGTTGGAGTC CACGTTCTTT
     5941 AATAGTGGAC TCTTGTTCCA AACTGGAACA ACACTCAACC CTATCTCGGT CT
//