LOCUS       sequence-317350-        5172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for tet-regulated sgRNA expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5172)
  AUTHORS   McGlincy NJ, Meacham ZA, Reynaud KK, Muller R, Baum R, Ingolia NT
  TITLE     A genome-scale CRISPR interference guide library enables 
            comprehensive phenotypic profiling in yeast.
  JOURNAL   BMC Genomics. 2021 Mar 23;22(1):205. doi: 
            10.1186/s12864-021-07518-0.
  PUBMED    33757429
REFERENCE   2  (bases 1 to 5172)
  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: "BMC 
            Genomics."; date: "2021-03-23"; volume: "22(1):205. doi"; pages: " 
            10.1186/s12864-021-07518-0"
COMMENT     Sequence Label: pNTI661 pRPR1(TetO)-sgRNA
FEATURES             Location/Qualifiers
     source          1..5172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        377..481
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(482..550,551..1342)
                     /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(700..719)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1513..2101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2002..2021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2496..2514
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        2590..2664
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      2677..2739
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
     primer_bind     complement(4950..4968)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(5009..5172,1..340)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa gaaaagcaga
       61 ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg attttcgtgt
      121 gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga gcaggaagag
      181 caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc ttcggaaaac
      241 aaaaactatt ttttctttaa tttctttttt tactttctat ttttaattta tatatttata
      301 ttaaaaaatt taaattataa ttatttttat agcacgtgat gaaaaggacc caggtggcac
      361 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      421 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      481 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      541 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
      601 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
      661 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
      721 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
      781 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
      841 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
      901 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
      961 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1021 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1081 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1141 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     1201 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1261 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1321 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1381 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1441 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1501 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1561 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1621 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1681 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1741 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1801 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1861 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1921 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1981 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2041 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2101 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctggcct
     2161 tttgctcggg ggatctgcca attgaacata acatggtagt tacatatact agtaatatgg
     2221 ttcggcacac attaaaagta taaaaactat ctgaattacg aattacatat attggtcata
     2281 aaaatcaatc aatcatcgtg tgttttatat gtctcttatc taagtataag aatatccata
     2341 gttaatattc acttacgcta ccttttaacc tgtaatcatt gtcaacagga tatgttaacg
     2401 acccacattg ataaacgcta gtatttcttt ttcctcttct tattggccgg ctgtctctat
     2461 actcccctat agtctgtttc ttttcgtttc gattgtccct atcagtgata gagatggcgc
     2521 acatggtacg ctgtggtgct cgcggctggg aacgaaactc tgggagctgc gattgggatc
     2581 ctcgaagctt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     2641 gaaaaagtgg caccgagtcg gtgctttttt ctcgagccat atccaacttc caatttaatc
     2701 tttctttttt aattttcact tatttgcgat acagaaagaa aaaagcgata gtaactattg
     2761 aattttgttt ggatttggtt agattagata tggtttctct ttatatttac atgctaaaaa
     2821 tgggctacac cagagataca taattagata tatatacgcc agtacacctt atcggcccaa
     2881 gccttgtccc aaggcagcgt tttgttcttg gaaacgctgc cctacacgtt cgctatgcgg
     2941 ccggtaccca gtttaaacga gctatcaggc gcgccacttc acgcatgctc aagagctcga
     3001 tccgcaggct aaccggaacc tgtattattt agtttatgct acgttaaata aagacctttc
     3061 gttcacataa ctgaatgtgt aatggccttg agatttcaag cataccaagt tggtggagac
     3121 ggggtcgtta caaaagactc tttaagccaa gatttccttg acagccttgg cgatagcatc
     3181 gccaacctca gtggtagagt tagaaccacc aaggtcaccg gttctgacac ctgcatccaa
     3241 gacatttcta acagcttctt caagagccct accttcttca accaaatcca aggataactt
     3301 caacatcata gctgcagata agatggtagc aattgggtta accttgtttg ctggtaaatc
     3361 tggggcagaa ccatgacatg gttcgtacaa accgaatgcc ttgttagtgt caggtaggga
     3421 agctagagat gcagaaggta ataaacccaa agaacctgga ataacagagg cttcatcgga
     3481 gataatatca ccaaacatgt tgttggtaat aacaacaccg tttagcttag ttggtgattt
     3541 aaccaaaatc atagcagcag agtcgatcaa ttggtgctga acagttaatt gtgggaactc
     3601 agtcttgatg gtttcttcaa cagtctttct ccacaatctg gaagaggcaa gcacgttagc
     3661 cttgtcaaga gaccagattg gtaatggtgg gttttgttgc aatgccaaga aagcagccat
     3721 tcttgtaatt ctttgaactt caggaacact gtatttctca gagtcccaag caactccgtc
     3781 accttcatct tcttttcttt caccaaagta gataccacca accaattctc taacgacgac
     3841 gaaatcggta ccctttgcat attcaggctt caaaggagaa agatctagta aagaatcaga
     3901 agcaaagtta catggtctca agttggcgta tagacccaat tcctttctga tcttcaatag
     3961 accttgttct ggtctaactg cgcccgtacc ccattttgga ccaccaacag cacctagtaa
     4021 gacagcatcg gctttcttag aggcttctag agcttcatct ggtaaaggag tgccagtggc
     4081 atcgatggca gcacccccga tcaagtgatg ttggaaattg aacttaattt ctggacggac
     4141 ttcagcaatg gcattcaaga ccttaatagc ttcgtcagta acttctttac cgacgtgatc
     4201 acccggtagg acaacgatat tcttagacat ggtataatct gtgtagtgtg ggatactttt
     4261 tacttcttca aataggtatc aacaataaaa ataaatcgaa gcaaatgtag gaatgcgtta
     4321 aagcagatgt acttacttag agtacataga tatatttata taattcaata tataaaagta
     4381 tatgaagcat ctgatgttga acctgtcatg actctaaact ggaatgggta gttatgcttc
     4441 tgaaggtttt ccgttagaaa atgagtgcag ggatcaggcc cggaaccggt tttagcctga
     4501 aaggaaaaga aaaaaatgcg gtgattacta gcacgtgact gcgctgaatt ggaatctacc
     4561 acaagtacag cgaatggcaa gagggaaggg gaatatattg tgcctggcgg tagcgtctgc
     4621 tatatcaaat gttaaacata actaaatgca gtagtgaata gagtaatgta tttctacata
     4681 tgtgggtagg gccaatggga gcccgatgtc tattgtacag atattttcct tttatagtta
     4741 atcaattcaa cttggaaatc attgagtcga tatctacttg gataactgga gccgtggcag
     4801 cgtttgaaat ttgacgagtc ttcttgtttt gttcgcgcat tttcagttta ttcaaacggg
     4861 agatggcacc gaatccactg acttctctgt tcaatttgaa tggtgaattc ggttttcacc
     4921 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4981 tgtcatgata ataatggttt cttagacgga tcgcttgcct gtaacttaca cgcgcctcgt
     5041 atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt ttatgttttg
     5101 tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga agaaaaaaaa
     5161 ataaacaaag gt
//
