LOCUS       sequence-371328-        5010 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 5010)
  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 5010)
  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-eDHFR-URA
FEATURES             Location/Qualifiers
     source          1..5010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(23..451)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(133..154)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     345..364
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     578..600
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     592..609
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     593..609
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     619..638
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        619..637
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     662..678
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     663..679
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             728..817
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             848..1327
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     promoter        1382..1725
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2544..2741
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2815..2835)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2815..2833)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2854..2870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2854..2870)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2867..2889)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2878..2894
                     /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(2902..2908,2909..2926,2927..2932))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2947..2968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3085..3102)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3256..3844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3336..3355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4015..4806,4807..4875))
                     /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     4638..4657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4876..4980)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
       61 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
      121 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
      181 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
      241 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
      301 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
      361 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
      421 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
      481 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      541 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      601 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
      661 ctcgaggtcg acggtatcga taagcttgat atcgggtcga cggatccccg ggttaattaa
      721 catcttttac ccatacgatg ttcctgacta tgcgggctat ccctatgacg tcccggacta
      781 tgcaggatcc tatccatatg acgttccaga ttacgctgct cagtgcggtt caggtggatc
      841 tggttctatg atcagtctga ttgcggcgtt agcggtagat cgcgttatcg gcatggaaaa
      901 cgccatgccg tggaacctgc ctgccgatct cgcctggttt aaacgcaaca ccttaaataa
      961 acccgtgatt atgggccgcc atacctggga atcaatcggt cgtccgttgc caggacgcaa
     1021 aaatattatc ctcagcagtc aaccgggtac ggacgatcgc gtaacgtggg tgaagtcggt
     1081 ggatgaagcc atcgcggcgt gtggtgacgt accagaaatc atggtgattg gcggcggtcg
     1141 cgtttatgaa cagttcttgc caaaagcgca aaaactgtat ctgacgcata tcgacgcaga
     1201 agtggaaggc gacacccatt tcccggatta cgagccggat gactgggaat cggtattcag
     1261 cgaattccac gatgctgatg cgcagaactc tcacagctat tgctttgaga ttctggagcg
     1321 gcggtaatat ataactgtct agaaataaag agtatcatct ttcaaaggtc acccggccag
     1381 cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg
     1441 atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat
     1501 acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga
     1561 gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag
     1621 aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat
     1681 cttgttagga tacagttctc acatcacatc cgaacataaa caaccatgac agtcaacact
     1741 aagacctata gtgagagagc agaaactcat gcctcaccag tagcacaacg attatttcga
     1801 ttaatggaac tgaagaaaac caatttatgt gcatcaattg atgttgatac cactaaggaa
     1861 ttccttgaat taattgataa attgggtcct tatgtatgct taatcaagac tcatattgat
     1921 ataatcaatg atttttccta tgaatccact attgaaccat tattagaact ttcacgtaaa
     1981 catcaattta tgatttttga agatagaaaa tttgctgata ttggtaatac cgtgaagaaa
     2041 caatatattg gtggagttta taaaattagt agttgggcag atattactaa tgctcatggt
     2101 gtcactggga atggagtagt tgaaggatta aaacagggag ctaaagaaac caccaccaac
     2161 caagagccaa gagggttatt gatgttagct gaattatcat cagtgggatc attagcatat
     2221 ggagaatatt ctcaaaaaac tgttgaaatt gctaaatccg ataaggaatt tgttattgga
     2281 tttattgccc aacgtgatat gggtggacaa gaagaaggat ttgattggct tattatgaca
     2341 cctggagttg gattagatga taaaggtgat ggattaggac aacaatatag aactgttgat
     2401 gaagttgtta gcactggaac tgatattatc attgttggta gaggattgtt tggtaaagga
     2461 agagatccag atattgaagg taaaaggtat agagatgctg gttggaatgc ttatttgaaa
     2521 aagactggcc aattataatc agtactgaca ataaaaagat tcttgttttc aagaacttgt
     2581 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     2641 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     2701 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac
     2761 gccgccacta gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta
     2821 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2881 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2941 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3001 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3061 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4981 cacatttccc cgaaaagtgc cacctaaatt
//
