LOCUS       sequence-181726-        3527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPRainbow-DONOR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3527)
  AUTHORS   Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T
  TITLE     Multiplexed labeling of genomic loci with dCas9 and engineered 
            sgRNAs using CRISPRainbow.
  JOURNAL   Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526.
  PUBMED    27088723
REFERENCE   2  (bases 1 to 3527)
  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: "Nat 
            Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526."
COMMENT     Sequence Label: pCRISPRainbow-DONOR1
FEATURES             Location/Qualifiers
     source          1..3527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(185..190,191..208,209..215)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             269..928
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(335..354)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1270..1575
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1454..1473
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(1688..2276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1768..1787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2447..3238,3239..3307))
                     /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     3070..3089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3308..3412)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3487..3506)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
       61 ggcaaggctt gaccgacaat tccatgaaga atctgcttag ggttaggcgt tttgcgctgc
      121 ttcgcgatgt acgggccaga tatacgcgtt gacagagacc gcggccgcat taggcacccc
      181 aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc
      241 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
      301 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
      361 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
      421 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
      481 ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
      541 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
      601 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
      661 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
      721 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
      781 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
      841 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
      901 ctgcgatgag tggcagggcg gggcgtaaag atctggatcc ggcttactaa aagccagata
      961 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
     1021 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
     1081 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     1141 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     1201 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     1261 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     1321 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
     1381 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     1441 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     1501 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     1561 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgacggtct
     1621 cgactcacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1681 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1741 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1801 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1861 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1921 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1981 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2041 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2101 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2161 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2221 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2281 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2341 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2401 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2461 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2521 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     2581 taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     2641 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2701 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2761 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2821 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2881 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2941 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3001 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3061 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3121 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3181 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3241 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3301 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3361 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     3421 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     3481 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgc
//
