LOCUS       sequence-371309-        5079 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 5079)
  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 5079)
  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-URA
FEATURES             Location/Qualifiers
     source          1..5079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(35..463)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(145..166)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     357..376
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     590..612
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     604..621
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     605..621
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     631..650
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        631..649
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     674..690
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     675..691
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             740..829
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             860..1405
                     /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"
     promoter        1463..1806
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2625..2822
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2896..2916)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2896..2914)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2935..2951)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2935..2951)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2948..2970)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2959..2975
                     /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(2983..2989,2990..3007,3008..3013))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3028..3049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3166..3183)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3337..3925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3417..3436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4096..4887,4888..4956))
                     /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     4719..4738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4957..5061)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
       61 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
      121 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
      181 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
      241 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
      301 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
      361 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
      421 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
      481 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
      541 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
      601 cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt
      661 accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgggtc gacggatccc
      721 cgggttaatt aacatctttt acccatacga tgttcctgac tatgcgggct atccctatga
      781 cgtcccggac tatgcaggat cctatccata tgacgttcca gattacgctg ctcagtgcgg
      841 ttcaggtgga tctggttcta tggacaaaga ctgcgaaatg aagcgcacca ccctggatag
      901 ccctctgggc aagctggaac tgtctgggtg cgaacagggc ctgcaccgta tcatcttcct
      961 gggcaaagga acatctgccg ccgacgccgt ggaagtgcct gccccagccg ccgtgctggg
     1021 cggaccagag ccactgatcc aggccaccgc ctggctcaac gcctactttc accagcctga
     1081 ggccatcgag gagttccctg tgccagccct gcaccaccca gtgttccagc aggagagctt
     1141 tacccgccag gtgctgtgga aactgctgaa agtggtgaag ttcggagagg tcatcagcga
     1201 gagccacctg gccgccctgg tgggcaatcc cgccgccacc gccgccgtga acaccgccct
     1261 ggacggaaat cccgtgccca ttctgatccc ctgccaccgg gtggtgcagg gcgacagcga
     1321 cgtggggccc tacctgggcg ggctcgccgt gaaagagtgg ctgctggccc acgagggcca
     1381 cagactgggc aagcctgggc tgggttaata tataactgtc tagaaataaa gagtatcatc
     1441 tttcaaaggt cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga
     1501 cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca
     1561 tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg
     1621 ctcctcgctg tagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc
     1681 ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt
     1741 tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa
     1801 acaaccatga cagtcaacac taagacctat agtgagagag cagaaactca tgcctcacca
     1861 gtagcacaac gattatttcg attaatggaa ctgaagaaaa ccaatttatg tgcatcaatt
     1921 gatgttgata ccactaagga attccttgaa ttaattgata aattgggtcc ttatgtatgc
     1981 ttaatcaaga ctcatattga tataatcaat gatttttcct atgaatccac tattgaacca
     2041 ttattagaac tttcacgtaa acatcaattt atgatttttg aagatagaaa atttgctgat
     2101 attggtaata ccgtgaagaa acaatatatt ggtggagttt ataaaattag tagttgggca
     2161 gatattacta atgctcatgg tgtcactggg aatggagtag ttgaaggatt aaaacaggga
     2221 gctaaagaaa ccaccaccaa ccaagagcca agagggttat tgatgttagc tgaattatca
     2281 tcagtgggat cattagcata tggagaatat tctcaaaaaa ctgttgaaat tgctaaatcc
     2341 gataaggaat ttgttattgg atttattgcc caacgtgata tgggtggaca agaagaagga
     2401 tttgattggc ttattatgac acctggagtt ggattagatg ataaaggtga tggattagga
     2461 caacaatata gaactgttga tgaagttgtt agcactggaa ctgatattat cattgttggt
     2521 agaggattgt ttggtaaagg aagagatcca gatattgaag gtaaaaggta tagagatgct
     2581 ggttggaatg cttatttgaa aaagactggc caattataat cagtactgac aataaaaaga
     2641 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     2701 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     2761 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     2821 tgctgtcgat tcgatactaa cgccgccact agttctagag cggccgccac cgcggtggag
     2881 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2941 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3001 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3061 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3121 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3181 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3241 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3301 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3361 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3421 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3481 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3541 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3601 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3661 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3721 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3781 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3841 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3901 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3961 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4021 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4081 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4141 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4201 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4261 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4321 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4381 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4441 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4501 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4561 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4621 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4681 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4741 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4801 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4861 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4921 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4981 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5041 taaacaaata ggggttccgc gcacatttcc ccgaaaagt
//
