LOCUS       sequence-371310-        5076 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 5076)
  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 5076)
  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-fCLIP-Kan
FEATURES             Location/Qualifiers
     source          1..5076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(115..183,184..975)
                     /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(333..352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1146..1734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1635..1654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1888..1905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2022..2043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2058..2063,2064..2081,2082..2088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2096..2112
                     /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     2101..2123
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2120..2136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2120..2136
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2155..2175
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2157..2175
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     gene            complement(2249..3605)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(2249..2446)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2452..3261)
                     /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     3175..3194
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3262..3605)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             complement(3663..4208)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylcytosine 
                     derivatives"
                     /label=CLIP-tag(TM)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             complement(4239..4328)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(4377..4393)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4378..4394)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4418..4437)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4419..4437)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4447..4464)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4447..4463)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4456..4478)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4605..5060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4692..4711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4902..4923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
       61 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      121 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      181 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      241 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      301 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      361 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
      421 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
      481 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
      541 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
      601 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
      661 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
      721 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
      781 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
      841 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
      901 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
      961 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1021 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1081 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1141 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1201 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1261 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1321 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1381 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1441 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1501 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1561 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1621 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1681 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1741 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     1801 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     1861 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     1921 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     1981 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2041 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2101 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2161 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctctagaact
     2221 agtggcggcg ttagtatcga atcgacagca gtatagcgac cagcattcac atacgattga
     2281 cgcatgatat tactttctgc gcacttaact tcgcatctgg gcagatgatg tcgaggcgaa
     2341 aaaaaatata aatcacgcta acatttgatt aaaatagaac aactacaata taaaaaaact
     2401 atacaaatga caagttcttg aaaacaagaa tctttttatt gtcagtactg attagaaaaa
     2461 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     2521 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     2581 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     2641 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     2701 tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     2761 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     2821 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     2881 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     2941 tacctggaat gctgttttgc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     3001 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     3061 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     3121 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     3181 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgaaac
     3241 gtgagtcttt tccttaccca tggttgttta tgttcggatg tgatgtgaga actgtatcct
     3301 agcaagattt taaaaggaag tatatgaaag aagaacctca gtggcaaatc ctaacctttt
     3361 atatttctct acaggggcgc ggcgtgggga caattcaacg cgtctgtgag gggagcgttt
     3421 ccctgctcgc aggtctgcag cgaggagccg taatttttgc ttcgcgccgt gcggccatca
     3481 aaatgtatgg atgcaaatga ttatacatgg ggatgtatgg gctaaatgta cgggcgacag
     3541 tcacatcatg cccctgagct gcgcacgtca agactgtcaa ggagggtatt ctgggcctcc
     3601 atgtcgctgg ccgggtgacc tttgaaagat gatactcttt atttctagac agttatatat
     3661 taacccagcc caggcttgcc cagtctgtgg ccctcgtggg ccagcagcca ctctttcacg
     3721 gcgagcccgc ccaggtaggg ccccacgtcg ctgtcgccct gcaccacccg gtggcagggg
     3781 atcagaatgg gcacgggatt tccgtccagg gcggtgttca cggcggcggt ggcggcggga
     3841 ttgcccacca gggcggccag gtggctctcg ctgatgacct ctccgaactt caccactttc
     3901 agcagtttcc acagcacctg gcgggtaaag ctctcctgct ggaacactgg gtggtgcagg
     3961 gctggcacag ggaactcctc gatggcctca ggctggtgaa agtaggcgtt gagccaggcg
     4021 gtggcctgga tcagtggctc tggtccgccc agcacggcgg ctggggcagg cacttccacg
     4081 gcgtcggcgg cagatgttcc tttgcccagg aagatgatac ggtgcaggcc ctgttcgcac
     4141 ccagacagtt ccagcttgcc cagagggcta tccagggtgg tgcgcttcat ttcgcagtct
     4201 ttgtccatag aaccagatcc acctgaaccg cactgagcag cgtaatctgg aacgtcatat
     4261 ggataggatc ctgcatagtc cgggacgtca tagggatagc ccgcatagtc aggaacatcg
     4321 tatgggtaaa agatgttaat taacccgggg atccgtcgac ccgatatcaa gcttatcgat
     4381 accgtcgacc tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc
     4441 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4501 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4561 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     4621 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4681 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4741 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4801 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4861 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4921 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4981 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5041 aatattaacg cttacaattt aggtggcact tttcgg
//
