LOCUS       sequence-371288-        4839 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 4839)
  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 4839)
  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-fSNAP-NAT
FEATURES             Location/Qualifiers
     source          1..4839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(18..473)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(155..176)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     367..386
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     600..622
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     614..631
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     615..631
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     641..660
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        641..659
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     684..700
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     685..701
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             750..839
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             870..1415
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     gene            1473..2592
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        1473..1816
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1817..2389
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      2395..2592
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2666..2686)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2666..2684)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2705..2721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2705..2721)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2718..2740)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2729..2745
                     /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(2753..2759,2760..2777,2778..2783))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2798..2819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2936..2953)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3107..3695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3187..3206)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3866..4657,4658..4726))
                     /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     4489..4508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4727..4831)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
       61 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
      121 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
      181 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
      241 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
      301 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
      361 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
      421 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
      481 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      541 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      601 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
      661 cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgggtc
      721 gacggatccc cgggttaatt aacatctttt acccatacga tgttcctgac tatgcgggct
      781 atccctatga cgtcccggac tatgcaggat cctatccata tgacgttcca gattacgctg
      841 ctcagtgcgg ttcaggtgga tctggttcta tggacaaaga ctgcgaaatg aagcgcacca
      901 ccctggatag ccctctgggc aagctggaac tgtctgggtg cgaacagggc ctgcaccgta
      961 tcatcttcct gggcaaagga acatctgccg ccgacgccgt ggaagtgcct gccccagccg
     1021 ccgtgctggg cggaccagag ccactgatgc aggccaccgc ctggctcaac gcctactttc
     1081 accagcctga ggccatcgag gagttccctg tgccagccct gcaccaccca gtgttccagc
     1141 aggagagctt tacccgccag gtgctgtgga aactgctgaa agtggtgaag ttcggagagg
     1201 tcatcagcta cagccacctg gccgccctgg ccggcaatcc cgccgccacc gccgccgtga
     1261 aaaccgccct gagcggaaat cccgtgccca ttctgatccc ctgccaccgg gtggtgcagg
     1321 gcgacctgga cgtggggggc tacgagggcg ggctcgccgt gaaagagtgg ctgctggccc
     1381 acgagggcca cagactgggc aagcctgggc tgggttaata tataactgtc tagaaataaa
     1441 gagtatcatc tttcaaaggt cacccggcca gcgacatgga ggcccagaat accctccttg
     1501 acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca
     1561 tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca
     1621 aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc acagacgcgt
     1681 tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga
     1741 ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat
     1801 ccgaacataa acaaccatgg gtaccactct tgacgacacg gcttaccggt accgcaccag
     1861 tgtcccgggg gacgccgagg ccatcgaggc actggatggg tccttcacca ccgacaccgt
     1921 cttccgcgtc accgccaccg gggacggctt caccctgcgg gaggtgccgg tggacccgcc
     1981 cctgaccaag gtgttccccg acgacgaatc ggacgacgaa tcggacgacg gggaggacgg
     2041 cgacccggac tcccggacgt tcgtcgcgta cggggacgac ggcgacctgg cgggcttcgt
     2101 ggtcgtctcg tactccggct ggaaccgccg gctgaccgtc gaggacatcg aggtcgcccc
     2161 ggagcaccgg gggcacgggg tcgggcgcgc gttgatgggg ctcgcgacgg agttcgcccg
     2221 cgagcggggc gccgggcacc tctggctgga ggtcaccaac gtcaacgcac cggcgatcca
     2281 cgcgtaccgg cggatggggt tcaccctctg cggcctggac accgccctgt acgacggcac
     2341 cgcctcggac ggcgagcagg cgctctacat gagcatgccc tgcccctaat cagtactgac
     2401 aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt
     2461 gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc
     2521 ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg
     2581 gtcgctatac tgctgtcgat tcgatactaa cgccgccact agttctagag cggccgccac
     2641 cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     2701 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2761 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2821 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2881 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2941 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3001 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3061 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3121 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3181 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3241 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3301 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3361 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3421 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3481 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3541 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3601 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3661 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3721 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3781 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3841 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3901 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3961 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4021 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4081 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4141 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4201 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4261 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4321 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4381 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4441 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4501 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4561 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4621 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4681 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4741 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4801 tttagaaaaa taaacaaata ggggttccgc gcacatttc
//
