LOCUS       sequence-371342-        5454 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 5454)
  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 5454)
  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-3xHA
FEATURES             Location/Qualifiers
     source          1..5454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..39)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(23..39)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(76..96)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(76..94)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(115..131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(115..131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(128..150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    139..155
                     /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(163..169,170..187,188..193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    208..229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(346..363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(517..1105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(597..616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1276..2067,2068..2136))
                     /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     1899..1918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2137..2241)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2267..2722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2404..2425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2616..2635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2849..2871
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2863..2880
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2864..2880
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2890..2909
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2890..2908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2985..3328
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4147..4344
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        4369..4810
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    4369..4486
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4781..4804
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4811..5356
                     /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"
     CDS             5357..5446
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
ORIGIN
        1 cggttcaggt ggatctggtt ctgatccact agttctagag cggccgccac cgcggtggag
       61 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
      121 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      181 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      241 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      301 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      361 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      421 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      481 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      541 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      601 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      661 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      721 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      781 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      841 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      901 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
      961 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1021 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1081 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1141 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1201 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1261 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1321 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1381 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1441 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1501 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1561 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1621 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1681 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1741 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1801 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1861 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1921 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1981 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2041 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2101 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2161 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2221 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     2281 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2341 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2401 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2461 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2521 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2581 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2641 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2701 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     2761 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     2821 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     2881 gagcgcgcgt aatacgactc actatagggc gaattgggta ccgggccccc cctcgaggtc
     2941 gacggtatag ataagcttga tatcgaattg gtcacccggc cagcgacatg gaggcccaga
     3001 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
     3061 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
     3121 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
     3181 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     3241 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     3301 ctcacatcac atccgaacat aaacaaccat gacagtcaac actaagacct atagtgagag
     3361 agcagaaact catgcctcac cagtagcaca acgattattt cgattaatgg aactgaagaa
     3421 aaccaattta tgtgcatcaa ttgatgttga taccactaag gaattccttg aattaattga
     3481 taaattgggt ccttatgtat gcttaatcaa gactcatatt gatataatca atgatttttc
     3541 ctatgaatcc actattgaac cattattaga actttcacgt aaacatcaat ttatgatttt
     3601 tgaagataga aaatttgctg atattggtaa taccgtgaag aaacaatata ttggtggagt
     3661 ttataaaatt agtagttggg cagatattac taatgctcat ggtgtcactg ggaatggagt
     3721 agttgaagga ttaaaacagg gagctaaaga aaccaccacc aaccaagagc caagagggtt
     3781 attgatgtta gctgaattat catcagtggg atcattagca tatggagaat attctcaaaa
     3841 aactgttgaa attgctaaat ccgataagga atttgttatt ggatttattg cccaacgtga
     3901 tatgggtgga caagaagaag gatttgattg gcttattatg acacctggag ttggattaga
     3961 tgataaaggt gatggattag gacaacaata tagaactgtt gatgaagttg ttagcactgg
     4021 aactgatatt atcattgttg gtagaggatt gtttggtaaa ggaagagatc cagatattga
     4081 aggtaaaagg tatagagatg ctggttggaa tgcttatttg aaaaagactg gccaattata
     4141 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     4201 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     4261 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     4321 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgccg gattagaagc
     4381 cgccgagcgg gcgacagccc tccgacggaa gactctcctc cgtgcgtcct cgtcttcacc
     4441 ggtcgcgttc ctgaaacgca gatgtgcctc gcgccgcact gctccgaaca ataaagattc
     4501 tacaatacta gcttttatgg ttatgaagag gaaaaattgg cagtaacctg gccccacaaa
     4561 ccttcaaatt aacgaatcaa attaacaacc ataggatgat aatgcgatta gttttttagc
     4621 cttatttctg gggtaattaa tcagcgaagc gatgattttt gatctattaa cagatatata
     4681 aatggaaaag ctgcataacc actttaacta atactttcaa cattttcagt ttgtattact
     4741 tcttattcaa atgtcataaa agtatcaaca aaaaattgtt aatatacctc tatactttaa
     4801 cgtcaaggag atggacaaag actgcgaaat gaagcgcacc accctggata gccctctggg
     4861 caagctggaa ctgtctgggt gcgaacaggg cctgcaccgt atcatcttcc tgggcaaagg
     4921 aacatctgcc gccgacgccg tggaagtgcc tgccccagcc gccgtgctgg gcggaccaga
     4981 gccactgatc caggccaccg cctggctcaa cgcctacttt caccagcctg aggccatcga
     5041 ggagttccct gtgccagccc tgcaccaccc agtgttccag caggagagct ttacccgcca
     5101 ggtgctgtgg aaactgctga aagtggtgaa gttcggagag gtcatcagcg agagccacct
     5161 ggccgccctg gtgggcaatc ccgccgccac cgccgccgtg aacaccgccc tggacggaaa
     5221 tcccgtgccc attctgatcc cctgccaccg ggtggtgcag ggcgacagcg acgtggggcc
     5281 ctacctgggc gggctcgccg tgaaagagtg gctgctggcc cacgagggcc acagactggg
     5341 caagcctggg ctgggttacc catacgatgt tcctgactat gcgggctatc cctatgacgt
     5401 cccggactat gcgggatcct atccatatga cgttccagat tacgctgctc agtg
//
