LOCUS       sequence-371345-        5355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration cassette to create N-terminal fusions for fluorescent 
            tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5355)
  AUTHORS   Baek I, Le SN, Jeon J, Chun Y, Reed C, Buratowski S
  TITLE     A set of Saccharomyces cerevisiae integration vectors for 
            fluorescent dye labeling of proteins.
  JOURNAL   G3 (Bethesda). 2022 Aug 9. pii: 6659097. doi: 
            10.1093/g3journal/jkac201.
  PUBMED    35944214
REFERENCE   2  (bases 1 to 5355)
  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: "G3 
            (Bethesda). 2022 Aug 9. pii: 6659097. doi: 
            10.1093/g3journal/jkac201."
COMMENT     Sequence Label: pBS-SKII-URA-GAL1-fCLIP-GS
FEATURES             Location/Qualifiers
     source          1..5355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..67)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(47..65)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(86..102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(86..102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(99..121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    110..126
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(134..140,141..158,159..164))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(317..334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(488..1076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(568..587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1247..2038,2039..2107))
                     /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     1870..1889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2108..2212)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2238..2693)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2375..2396)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2587..2606
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2820..2842
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2834..2851
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2835..2851
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2861..2880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2861..2879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2956..3299
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4118..4315
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        4340..4781
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    4340..4457
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4752..4775
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4782..5327
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylcytosine 
                     derivatives"
                     /label=CLIP-tag(TM)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLGGLA
                     VKEWLLAHEGHRLGKPGLG"
     primer_bind     complement(join(5349..5355,1..10))
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(5349..5355,1..10))
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
       61 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      121 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      181 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      241 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
      301 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      361 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      421 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      481 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      661 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      841 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      901 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      961 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1021 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1081 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1141 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1201 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1261 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1321 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1381 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1441 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1501 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1561 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1621 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1681 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1741 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1801 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1861 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1921 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1981 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2041 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2101 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2161 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2221 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2281 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2341 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2401 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2461 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2521 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2581 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2641 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     2701 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2761 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2821 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     2881 cgaattgggt accgggcccc ccctcgaggt cgacggtata gataagcttg atatcgaatt
     2941 ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc
     3001 agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa
     3061 tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg
     3121 ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc
     3181 gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata
     3241 cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca
     3301 tgacagtcaa cactaagacc tatagtgaga gagcagaaac tcatgcctca ccagtagcac
     3361 aacgattatt tcgattaatg gaactgaaga aaaccaattt atgtgcatca attgatgttg
     3421 ataccactaa ggaattcctt gaattaattg ataaattggg tccttatgta tgcttaatca
     3481 agactcatat tgatataatc aatgattttt cctatgaatc cactattgaa ccattattag
     3541 aactttcacg taaacatcaa tttatgattt ttgaagatag aaaatttgct gatattggta
     3601 ataccgtgaa gaaacaatat attggtggag tttataaaat tagtagttgg gcagatatta
     3661 ctaatgctca tggtgtcact gggaatggag tagttgaagg attaaaacag ggagctaaag
     3721 aaaccaccac caaccaagag ccaagagggt tattgatgtt agctgaatta tcatcagtgg
     3781 gatcattagc atatggagaa tattctcaaa aaactgttga aattgctaaa tccgataagg
     3841 aatttgttat tggatttatt gcccaacgtg atatgggtgg acaagaagaa ggatttgatt
     3901 ggcttattat gacacctgga gttggattag atgataaagg tgatggatta ggacaacaat
     3961 atagaactgt tgatgaagtt gttagcactg gaactgatat tatcattgtt ggtagaggat
     4021 tgtttggtaa aggaagagat ccagatattg aaggtaaaag gtatagagat gctggttgga
     4081 atgcttattt gaaaaagact ggccaattat aatcagtact gacaataaaa agattcttgt
     4141 tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa
     4201 tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag
     4261 tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc
     4321 gattcgatac taacgccgcc ggattagaag ccgccgagcg ggcgacagcc ctccgacgga
     4381 agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct
     4441 cgcgccgcac tgctccgaac aataaagatt ctacaatact agcttttatg gttatgaaga
     4501 ggaaaaattg gcagtaacct ggccccacaa accttcaaat taacgaatca aattaacaac
     4561 cataggatga taatgcgatt agttttttag ccttatttct ggggtaatta atcagcgaag
     4621 cgatgatttt tgatctatta acagatatat aaatggaaaa gctgcataac cactttaact
     4681 aatactttca acattttcag tttgtattac ttcttattca aatgtcataa aagtatcaac
     4741 aaaaaattgt taatatacct ctatacttta acgtcaagga gatggacaaa gactgcgaaa
     4801 tgaagcgcac caccctggat agccctctgg gcaagctgga actgtctggg tgcgaacagg
     4861 gcctgcaccg tatcatcttc ctgggcaaag gaacatctgc cgccgacgcc gtggaagtgc
     4921 ctgccccagc cgccgtgctg ggcggaccag agccactgat ccaggccacc gcctggctca
     4981 acgcctactt tcaccagcct gaggccatcg aggagttccc tgtgccagcc ctgcaccacc
     5041 cagtgttcca gcaggagagc tttacccgcc aggtgctgtg gaaactgctg aaagtggtga
     5101 agttcggaga ggtcatcagc gagagccacc tggccgccct ggtgggcaat cccgccgcca
     5161 ccgccgccgt gaacaccgcc ctggacggaa atcccgtgcc cattctgatc ccctgccacc
     5221 gggtggtgca gggcgacagc gacgtggggc cctacctggg cgggctcgcc gtgaaagagt
     5281 ggctgctggc ccacgagggc cacagactgg gcaagcctgg gctgggtggt tcaggtggat
     5341 ctggttctga tccac
//
