LOCUS       sequence-371329-        5007 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 5007)
  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 5007)
  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-Kan
FEATURES             Location/Qualifiers
     source          1..5007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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)."
     primer_bind     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     648..668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        650..668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     697..713
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     698..714
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             763..852
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             883..1362
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     gene            1417..2773
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1417..1760
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1761..2570
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1828..1847)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2576..2773
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2842..2861)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2843..2861)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2871..2888)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2871..2887)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2880..2902)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3029..3484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3116..3135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3326..3347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3510..3614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3615..3683,3684..4475)
                     /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     complement(3833..3852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4646..5007,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgctcccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      661 actaaaggga acaaaagctg ggtaccgggc cccccctcga ggtcgacggt atcgataagc
      721 ttgatatcgg gtcgacggat ccccgggtta attaacatct tttacccata cgatgttcct
      781 gactatgcgg gctatcccta tgacgtcccg gactatgcag gatcctatcc atatgacgtt
      841 ccagattacg ctgctcagtg cggttcaggt ggatctggtt ctatgatcag tctgattgcg
      901 gcgttagcgg tagatcgcgt tatcggcatg gaaaacgcca tgccgtggaa cctgcctgcc
      961 gatctcgcct ggtttaaacg caacacctta aataaacccg tgattatggg ccgccatacc
     1021 tgggaatcaa tcggtcgtcc gttgccagga cgcaaaaata ttatcctcag cagtcaaccg
     1081 ggtacggacg atcgcgtaac gtgggtgaag tcggtggatg aagccatcgc ggcgtgtggt
     1141 gacgtaccag aaatcatggt gattggcggc ggtcgcgttt atgaacagtt cttgccaaaa
     1201 gcgcaaaaac tgtatctgac gcatatcgac gcagaagtgg aaggcgacac ccatttcccg
     1261 gattacgagc cggatgactg ggaatcggta ttcagcgaat tccacgatgc tgatgcgcag
     1321 aactctcaca gctattgctt tgagattctg gagcggcggt aatatataac tgtctagaaa
     1381 taaagagtat catctttcaa aggtcacccg gccagcgaca tggaggccca gaataccctc
     1441 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     1501 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     1561 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     1621 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     1681 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     1741 acatccgaac ataaacaacc atgggtaagg aaaagactca cgtttcgagg ccgcgattaa
     1801 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     1861 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
     1921 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     1981 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     2041 tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa tatcctgatt
     2101 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     2161 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     2221 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     2281 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc
     2341 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     2401 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     2461 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     2521 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagtactga
     2581 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt
     2641 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg
     2701 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct
     2761 ggtcgctata ctgctgtcga ttcgatacta acgccgccac tagttctaga gcggccgcca
     2821 ccgcggtgga gctccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     2881 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2941 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3001 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     3061 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3121 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3181 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3241 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3301 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3361 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3421 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3481 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3541 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3601 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3661 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3721 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3781 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3841 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3901 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3961 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4021 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4081 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4141 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4261 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4321 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4441 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4501 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4561 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4621 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4681 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4741 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4981 cacagcccag cttggagcga acgacct
//
