LOCUS       sequence-371340-        5385 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 5385)
  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 5385)
  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-DHFR-3xHA
FEATURES             Location/Qualifiers
     source          1..5385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..31)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15..31)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(68..88)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(68..86)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(107..123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(107..123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(120..142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    131..147
                     /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(155..161,162..179,180..185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    200..221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(338..355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(509..1097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(589..608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1268..2059,2060..2128))
                     /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     1891..1910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2129..2233)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2259..2714)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2396..2417)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2608..2627
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2841..2863
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2855..2872
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2856..2872
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2882..2901
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2882..2900
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2977..3320
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4139..4336
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        4361..4802
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    4361..4478
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4773..4796
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             4803..5279
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             5280..5369
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
ORIGIN
        1 gtggatctgg ttctgatcca ctagttctag agcggccgcc accgcggtgg agctccagct
       61 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
      121 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
      181 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      241 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      301 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      361 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      421 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      481 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      541 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      601 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      661 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      721 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      781 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      841 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      901 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
      961 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1021 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1081 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1141 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1201 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1261 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1321 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1381 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1441 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1501 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1561 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1621 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1681 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     1741 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1801 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     1861 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     1921 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     1981 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2041 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2101 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2161 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2221 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     2281 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2341 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2401 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2461 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     2521 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2581 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2641 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2701 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     2761 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     2821 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc
     2881 gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg tcgacggtat
     2941 agataagctt gatatcgaat tggtcacccg gccagcgaca tggaggccca gaataccctc
     3001 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     3061 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     3121 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     3181 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     3241 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     3301 acatccgaac ataaacaacc atgacagtca acactaagac ctatagtgag agagcagaaa
     3361 ctcatgcctc accagtagca caacgattat ttcgattaat ggaactgaag aaaaccaatt
     3421 tatgtgcatc aattgatgtt gataccacta aggaattcct tgaattaatt gataaattgg
     3481 gtccttatgt atgcttaatc aagactcata ttgatataat caatgatttt tcctatgaat
     3541 ccactattga accattatta gaactttcac gtaaacatca atttatgatt tttgaagata
     3601 gaaaatttgc tgatattggt aataccgtga agaaacaata tattggtgga gtttataaaa
     3661 ttagtagttg ggcagatatt actaatgctc atggtgtcac tgggaatgga gtagttgaag
     3721 gattaaaaca gggagctaaa gaaaccacca ccaaccaaga gccaagaggg ttattgatgt
     3781 tagctgaatt atcatcagtg ggatcattag catatggaga atattctcaa aaaactgttg
     3841 aaattgctaa atccgataag gaatttgtta ttggatttat tgcccaacgt gatatgggtg
     3901 gacaagaaga aggatttgat tggcttatta tgacacctgg agttggatta gatgataaag
     3961 gtgatggatt aggacaacaa tatagaactg ttgatgaagt tgttagcact ggaactgata
     4021 ttatcattgt tggtagagga ttgtttggta aaggaagaga tccagatatt gaaggtaaaa
     4081 ggtatagaga tgctggttgg aatgcttatt tgaaaaagac tggccaatta taatcagtac
     4141 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     4201 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     4261 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     4321 gctggtcgct atactgctgt cgattcgata ctaacgccgc cggattagaa gccgccgagc
     4381 gggcgacagc cctccgacgg aagactctcc tccgtgcgtc ctcgtcttca ccggtcgcgt
     4441 tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa caataaagat tctacaatac
     4501 tagcttttat ggttatgaag aggaaaaatt ggcagtaacc tggccccaca aaccttcaaa
     4561 ttaacgaatc aaattaacaa ccataggatg ataatgcgat tagtttttta gccttatttc
     4621 tggggtaatt aatcagcgaa gcgatgattt ttgatctatt aacagatata taaatggaaa
     4681 agctgcataa ccactttaac taatactttc aacattttca gtttgtatta cttcttattc
     4741 aaatgtcata aaagtatcaa caaaaaattg ttaatatacc tctatacttt aacgtcaagg
     4801 agatgatcag tctgattgcg gcgttagcgg tagatcgcgt tatcggcatg gaaaacgcca
     4861 tgccgtggaa cctgcctgcc gatctcgcct ggtttaaacg caacacctta aataaacccg
     4921 tgattatggg ccgccatacc tgggaatcaa tcggtcgtcc gttgccagga cgcaaaaata
     4981 ttatcctcag cagtcaaccg ggtacggacg atcgcgtaac gtgggtgaag tcggtggatg
     5041 aagccatcgc ggcgtgtggt gacgtaccag aaatcatggt gattggcggc ggtcgcgttt
     5101 atgaacagtt cttgccaaaa gcgcaaaaac tgtatctgac gcatatcgac gcagaagtgg
     5161 aaggcgacac ccatttcccg gattacgagc cggatgactg ggaatcggta ttcagcgaat
     5221 tccacgatgc tgatgcgcag aactctcaca gctattgctt tgagattctg gagcggcggt
     5281 acccatacga tgttcctgac tatgcgggct atccctatga cgtcccggac tatgcgggat
     5341 cctatccata tgacgttcca gattacgctg ctcagtgcgg ttcag
//
