LOCUS       sequence-371287-        4652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration cassette to create C-terminal fusions for fluorescent 
            tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4652)
  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 4652)
  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-3XHA-fSNAP-Phleomycin
FEATURES             Location/Qualifiers
     source          1..4652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(15..470)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(152..173)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     364..383
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     597..619
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     611..628
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     612..628
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     681..697
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     682..698
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             746..835
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             866..1411
                     /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"
     promoter        1469..1812
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2205..2402
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2476..2496)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2476..2494)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2515..2531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2515..2531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2528..2550)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2539..2555
                     /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(2563..2569,2570..2587,2588..2593))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2608..2629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2746..2763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2917..3505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2997..3016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3676..4467,4468..4536))
                     /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     4299..4318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4537..4641)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
       61 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
      121 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
      181 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
      241 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
      301 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
      361 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
      421 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
      481 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
      541 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
      601 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
      661 attgggtacc gggccccccc tcgaggtcga cggtatcgat aagttgatat cgggtcgacg
      721 gatccccggg ttaattaaca tcttttaccc atacgatgtt cctgactatg cgggctatcc
      781 ctatgacgtc ccggactatg caggatccta tccatatgac gttccagatt acgctgctca
      841 gtgcggttca ggtggatctg gttctatgga caaagactgc gaaatgaagc gcaccaccct
      901 ggatagccct ctgggcaagc tggaactgtc tgggtgcgaa cagggcctgc accgtatcat
      961 cttcctgggc aaaggaacat ctgccgccga cgccgtggaa gtgcctgccc cagccgccgt
     1021 gctgggcgga ccagagccac tgatgcaggc caccgcctgg ctcaacgcct actttcacca
     1081 gcctgaggcc atcgaggagt tccctgtgcc agccctgcac cacccagtgt tccagcagga
     1141 gagctttacc cgccaggtgc tgtggaaact gctgaaagtg gtgaagttcg gagaggtcat
     1201 cagctacagc cacctggccg ccctggccgg caatcccgcc gccaccgccg ccgtgaaaac
     1261 cgccctgagc ggaaatcccg tgcccattct gatcccctgc caccgggtgg tgcagggcga
     1321 cctggacgtg gggggctacg agggcgggct cgccgtgaaa gagtggctgc tggcccacga
     1381 gggccacaga ctgggcaagc ctgggctggg ttaatatata actgtctaga aataaagagt
     1441 atcatctttc aaaggtcacc cggccagcga catggaggcc cagaataccc tccttgacag
     1501 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
     1561 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
     1621 ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
     1681 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
     1741 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
     1801 acataaacaa ccatgggtat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     1861 gatcccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     1921 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc cgggctcgat
     1981 cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg
     2041 cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc
     2101 gaactgcagg agtggggagg cacgatggcc gctttggtcg acccggacgg gacgctcctg
     2161 cgcctgatac agaacgaatt gcttgcaggc atctcatgat cagtactgac aataaaaaga
     2221 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     2281 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     2341 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     2401 tgctgtcgat tcgatactaa cgccgccact agttctagag cggccgccac cgcggtggag
     2461 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2521 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2581 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2641 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2701 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2761 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2821 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3661 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3721 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3781 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3841 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3901 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3961 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4021 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4081 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4141 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4201 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4261 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4321 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4381 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4441 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4501 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4561 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4621 taaacaaata ggggttccgc gcacatttcc cc
//
