LOCUS       sequence-381551-        5421 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 5421)
  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 5421)
  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-HALO-Kanamycin
FEATURES             Location/Qualifiers
     source          1..5421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(14..469)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(151..172)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     363..382
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     596..618
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     610..627
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     611..627
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     637..656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        637..655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     680..696
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     681..697
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             746..835
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             866..1756
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     gene            1814..3170
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1814..2157
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2158..2967
                     /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(2225..2244)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2973..3170
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3244..3264)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3244..3262)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3283..3299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3283..3299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3296..3318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3307..3323
                     /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(3331..3337,3338..3355,3356..3361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3376..3397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3514..3531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3685..4273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3765..3784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4444..5235,5236..5304))
                     /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     5067..5086
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5305..5409)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
       61 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
      121 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
      181 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
      241 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
      301 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
      361 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
      421 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
      481 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      541 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
      601 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
      661 ttgggtaccg ggccccccct cgaggtcgac ggtatcgata agcttgatat cgggtcgacg
      721 gatccccggg ttaattaaca tcttttaccc atacgatgtt cctgactatg cgggctatcc
      781 ctatgacgtc ccggactatg caggatccta tccatatgac gttccagatt acgctgctca
      841 gtgcggttca ggtggatctg gttctatggc agaaatcggt actggctttc cattcgaccc
      901 gcattatgtg gaagtcctgg gcgagagaat gcattacgtt gacgtgggtc cgagagatgg
      961 aactccggtc ctttttctgc acgggaatcc tacaagctct tatgtttggc gcaatatcat
     1021 ccctcatgta gctccgacgc atcgctgtat tgcgccggac ctgattggta tgggaaaatc
     1081 tgataaacca gacctgggtt actttttcga tgatcatgtg cgtttcatgg atgccttcat
     1141 tgaggcatta gggcttgaag aagtcgtctt ggtgattcat gattggggct cagctctggg
     1201 atttcactgg gctaaaagaa atcctgaacg cgtaaaaggc atcgcgttta tggagttcat
     1261 tcgtccaatt ccgacttggg atgaatggcc tgagttcgcg agagaaacat ttcaagcatt
     1321 tcgcacgacc gatgtaggcc ggaagttaat catcgatcag aatgtcttta tcgaagggac
     1381 attgccgatg ggagtcgttc gtccgttaac agaagtcgaa atggatcact atagagaacc
     1441 ttttcttaat cctgtggaca gagagccgct gtggcggttt ccgaacgaac tgccgattgc
     1501 aggcgagcct gctaacattg tagcgctggt tgaagagtat atggattggc ttcatcagtc
     1561 tccagttccg aagttattgt tttggggtac gcctggcgtg cttattccac cggccgaagc
     1621 ggcacgtttg gcaaaaagcc tgccaaattg caaagccgtt gacattggcc ctggacttaa
     1681 cttgcttcaa gaggataacc cggacttaat cgggagcgaa attgcccggt ggctttctac
     1741 cttagaaatc agcggctaat atataactgt ctagaaataa agagtatcat ctttcaaagg
     1801 tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag
     1861 ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc
     1921 atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct
     1981 gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc
     2041 gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact
     2101 tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccatg
     2161 ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga tgctgattta
     2221 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
     2281 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     2341 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     2401 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc
     2461 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     2521 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     2581 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     2641 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     2701 aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     2761 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     2821 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     2881 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     2941 catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt cttgttttca
     3001 agaacttgtc atttgtatag tttttttata ttgtagttgc tctattttaa tcaaatgtta
     3061 gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc
     3121 agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc
     3181 gatactaacg ccgccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg
     3241 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     3301 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3361 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3421 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3481 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3601 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3661 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3721 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3781 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3841 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3901 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3961 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4021 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4081 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4141 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4201 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4261 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4321 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4381 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4441 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4501 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4561 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4621 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4681 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4741 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4801 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4861 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4921 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4981 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5041 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5101 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5161 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5221 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5281 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5341 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5401 ggttccgcgc acatttcccc g
//
