LOCUS       sequence-381552-        5354 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 5354)
  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 5354)
  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-fSNAP-GS
FEATURES             Location/Qualifiers
     source          1..5354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..48)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(32..48)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(85..105)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(85..103)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(124..140)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(124..140)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(137..159)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    148..164
                     /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(172..178,179..196,197..202))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    217..238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(355..372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(526..1114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(606..625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1285..2076,2077..2145))
                     /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     1908..1927
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2146..2250)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2276..2731)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2413..2434)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2625..2644
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2858..2880
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2872..2889
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2873..2889
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2899..2918
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2899..2917
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2994..3337
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4156..4352
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        4377..4818
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    4377..4494
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4789..4812
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             join(4819..5354,1..10)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
ORIGIN
        1 tgggctgggt ggttcaggtg gatctggttc tgatccacta gttctagagc ggccgccacc
       61 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
      121 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      181 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      241 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      301 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      361 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      421 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      481 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      541 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      601 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      661 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      721 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      781 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      841 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      901 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      961 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1021 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1081 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1141 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1201 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1261 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1321 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1381 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1441 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1501 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1561 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1621 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1681 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1741 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1801 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1861 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1921 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1981 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2041 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2101 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2161 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2221 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     2281 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2341 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2401 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2461 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2521 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2581 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2641 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2701 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     2761 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     2821 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     2881 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
     2941 ctcgaggtcg acggtataga taagcttgat atcgaattgg tcacccggcc agcgacatgg
     3001 aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca tgatgtgact
     3061 gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc atacattttg
     3121 atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc gagcagggaa
     3181 acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag agaaatataa
     3241 aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa atcttgctag
     3301 gatacagttc tcacatcaca tccgaacata aacaaccatg acagtcaaca ctaagaccta
     3361 tagtgagaga gcagaaactc atgcctcacc agtagcacaa cgattatttc gattaatgga
     3421 actgaagaaa accaatttat gtgcatcaat tgatgttgat accactaagg aattccttga
     3481 attaattgat aaattgggtc cttatgtatg cttaatcaag actcatattg atataatcaa
     3541 tgatttttcc tatgaatcca ctattgaacc attattagaa ctttcacgta aacatcaatt
     3601 tatgattttt gaagatagaa aatttgctga tattggtaat accgtgaaga aacaatatat
     3661 tggtggagtt tataaaatta gtagttgggc agatattact aatgctcatg gtgtcactgg
     3721 gaatggagta gttgaaggat taaaacaggg agctaaagaa accaccacca accaagagcc
     3781 aagagggtta ttgatgttag ctgaattatc atcagtggga tcattagcat atggagaata
     3841 ttctcaaaaa actgttgaaa ttgctaaatc cgataaggaa tttgttattg gatttattgc
     3901 ccaacgtgat atgggtggac aagaagaagg atttgattgg cttattatga cacctggagt
     3961 tggattagat gataaaggtg atggattagg acaacaatat agaactgttg atgaagttgt
     4021 tagcactgga actgatatta tcattgttgg tagaggattg tttggtaaag gaagagatcc
     4081 agatattgaa ggtaaaaggt atagagatgc tggttggaat gcttatttga aaaagactgg
     4141 ccaattataa tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     4201 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     4261 ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg
     4321 tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccgga
     4381 ttagaagccg ccgagcgggc gacagccctc cgacggaaga ctctcctccg tgcgtcctcg
     4441 tcttcaccgg tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat
     4501 aaagattcta caatactagc ttttatggtt atgaagagga aaaattggca gtaacctggc
     4561 cccacaaacc ttcaaattaa cgaatcaaat taacaaccat aggatgataa tgcgattagt
     4621 tttttagcct tatttctggg gtaattaatc agcgaagcga tgatttttga tctattaaca
     4681 gatatataaa tggaaaagct gcataaccac tttaactaat actttcaaca ttttcagttt
     4741 gtattacttc ttattcaaat gtcataaaag tatcaacaaa aaattgttaa tatacctcta
     4801 tactttaacg tcaaggagat ggacaaagac tgcgaaatga agcgcaccac cctggatagc
     4861 cctctgggca agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg
     4921 ggcaaaggaa catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc
     4981 ggaccagagc cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag
     5041 gccatcgagg agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt
     5101 acccgccagg tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac
     5161 agccacctgg ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg
     5221 agcggaaatc ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacctggac
     5281 gtggggggct acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac
     5341 agactgggca agcc
//
