LOCUS       sequence-371289-        5078 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 5078)
  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 5078)
  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-URA
FEATURES             Location/Qualifiers
     source          1..5078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(27..482)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(164..185)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     376..395
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     609..631
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     623..640
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     624..640
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     650..669
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        650..668
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     693..709
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     694..710
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             759..848
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             879..1424
                     /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        1482..1825
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2644..2841
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2914..2934)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2914..2932)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2953..2969)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2953..2969)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2966..2988)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2977..2993
                     /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(3001..3007,3008..3025,3026..3031))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3046..3067
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3184..3201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3355..3943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3435..3454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4114..4905,4906..4974))
                     /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     4737..4756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4975..5078,1))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
       61 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
      121 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
      181 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
      241 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
      301 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
      361 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
      421 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
      481 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
      541 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      601 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
      661 actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga
      721 tatcgggtcg acggatcccc gggttaatta acatctttta cccatacgat gttcctgact
      781 atgcgggcta tccctatgac gtcccggact atgcgggatc ctatccatat gacgttccag
      841 attacgctgc tcagtgcggt tcaggtggat ctggttctat ggacaaagac tgcgaaatga
      901 agcgcaccac cctggatagc cctctgggca agctggaact gtctgggtgc gaacagggcc
      961 tgcaccgtat catcttcctg ggcaaaggaa catctgccgc cgacgccgtg gaagtgcctg
     1021 ccccagccgc cgtgctgggc ggaccagagc cactgatgca ggccaccgcc tggctcaacg
     1081 cctactttca ccagcctgag gccatcgagg agttccctgt gccagccctg caccacccag
     1141 tgttccagca ggagagcttt acccgccagg tgctgtggaa actgctgaaa gtggtgaagt
     1201 tcggagaggt catcagctac agccacctgg ccgccctggc cggcaatccc gccgccaccg
     1261 ccgccgtgaa aaccgccctg agcggaaatc ccgtgcccat tctgatcccc tgccaccggg
     1321 tggtgcaggg cgacctggac gtggggggct acgagggcgg gctcgccgtg aaagagtggc
     1381 tgctggccca cgagggccac agactgggca agcctgggct gggttaatat ataactgtct
     1441 agaaataaag agtatcatct ttcaaaggtc acccggccag cgacatggag gcccagaata
     1501 ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca
     1561 tttagcccat acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg
     1621 cgcgaagcaa aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca
     1681 cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt
     1741 tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc
     1801 acatcacatc cgaacataaa caaccatgac agtcaacact aagacctata gtgagagagc
     1861 agaaactcat gcctcaccag tagcacaacg attatttcga ttaatggaac tgaagaaaac
     1921 caatttatgt gcatcaattg atgttgatac cactaaggaa ttccttgaat taattgataa
     1981 attgggtcct tatgtatgct taatcaagac tcatattgat ataatcaatg atttttccta
     2041 tgaatccact attgaaccat tattagaact ttcacgtaaa catcaattta tgatttttga
     2101 agatagaaaa tttgctgata ttggtaatac cgtgaagaaa caatatattg gtggagttta
     2161 taaaattagt agttgggcag atattactaa tgctcatggt gtcactggga atggagtagt
     2221 tgaaggatta aaacagggag ctaaagaaac caccaccaac caagagccaa gagggttatt
     2281 gatgttagct gaattatcat cagtgggatc attagcatat ggagaatatt ctcaaaaaac
     2341 tgttgaaatt gctaaatccg ataaggaatt tgttattgga tttattgccc aacgtgatat
     2401 gggtggacaa gaagaaggat ttgattggct tattatgaca cctggagttg gattagatga
     2461 taaaggtgat ggattaggac aacaatatag aactgttgat gaagttgtta gcactggaac
     2521 tgatattatc attgttggta gaggattgtt tggtaaagga agagatccag atattgaagg
     2581 taaaaggtat agagatgctg gttggaatgc ttatttgaaa aagactggcc aattataatc
     2641 agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat
     2701 attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac
     2761 atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg
     2821 tgaatgctgg tcgctatact gctgtcgatt cgatactaac gcgccactag ttctagagcg
     2881 gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     2941 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3001 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3061 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3121 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3181 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3241 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3301 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3361 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3421 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3481 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3541 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3601 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3661 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3721 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3781 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3841 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3901 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3961 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4021 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4081 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4141 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4201 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4261 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4321 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4381 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4441 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4501 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4561 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4621 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4681 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4741 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4801 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4861 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4921 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4981 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5041 tgaatgtatt tagaaaaata aacaaatagg ggttccgc
//
