LOCUS       sequence-371291-        5295 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 5295)
  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 5295)
  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-Hygromycin
FEATURES             Location/Qualifiers
     source          1..5295
                     /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"
     gene            1463..3038
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        1463..1806
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1807..2835
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             1807..2835
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      2841..3038
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3112..3132)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3112..3130)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3151..3167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3151..3167)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3164..3186)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3175..3191
                     /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(3199..3205,3206..3223,3224..3229))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3244..3265
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3382..3399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3553..4141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3633..3652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4312..5103,5104..5172))
                     /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     4935..4954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5173..5277)
                     /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 cggacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc
     1681 ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt
     1741 tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa
     1801 acaaccatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     1861 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     1921 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     1981 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     2041 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     2101 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     2161 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     2221 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     2281 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     2341 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     2401 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     2461 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     2521 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     2581 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     2641 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     2701 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     2761 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     2821 agggcaaagg aataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     2881 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     2941 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     3001 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     3061 gccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg
     3121 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3181 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3241 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3301 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3361 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3421 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3481 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3541 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3601 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3661 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3721 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3781 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3841 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3901 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3961 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4021 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4081 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4141 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4201 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4261 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4321 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4381 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4441 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4501 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4561 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4621 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4681 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4741 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4801 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4861 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4921 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4981 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5041 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5101 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5161 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5221 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5281 atttccccga aaagt
//
