LOCUS       sequence-371324-        4584 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 4584)
  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 4584)
  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-eDHFR-Phleomycin
FEATURES             Location/Qualifiers
     source          1..4584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             735..824
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             855..1334
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     promoter        1389..1732
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2125..2322
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2396..2416)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2396..2414)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2435..2451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2435..2451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2448..2470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2459..2475
                     /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(2483..2489,2490..2507,2508..2513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2528..2549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2666..2683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2837..3425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2917..2936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3596..4387,4388..4456))
                     /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     4219..4238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4457..4561)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gggtcgacgg atccccgggt
      721 taattaacat cttttaccca tacgatgttc ctgactatgc gggctatccc tatgacgtcc
      781 cggactatgc aggatcctat ccatatgacg ttccagatta cgctgctcag tgcggttcag
      841 gtggatctgg ttctatgatc agtctgattg cggcgttagc ggtagatcgc gttatcggca
      901 tggaaaacgc catgccgtgg aacctgcctg ccgatctcgc ctggtttaaa cgcaacacct
      961 taaataaacc cgtgattatg ggccgccata cctgggaatc aatcggtcgt ccgttgccgg
     1021 gacgcaaaaa tattatcctc agcagtcaac cgggtacgga cgatcgcgta acgtgggtga
     1081 agtcggtgga tgaagccatc gcggcgtgtg gtgacgtacc agaaatcatg gtgattggcg
     1141 gcggtcgcgt ttatgaacag ttcttgccaa aagcgcaaaa actgtatctg acgcatatcg
     1201 acgcagaagt ggaaggcgac acccatttcc cggattacga gccggatgac tgggaatcgg
     1261 tattcagcga attccacgat gctgatgcgc agaactctca cagctattgc tttgagattc
     1321 tggagcggcg gtaatatata actgtctaga aataaagagt atcatctttc aaaggtcacc
     1381 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
     1441 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
     1501 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga
     1561 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
     1621 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
     1681 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtat
     1741 gaccgaccaa gcgacgccca acctgccatc acgagatttc gatcccaccg ccgccttcta
     1801 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     1861 ggatctcatg ctggagttct tcgcccaccc cgggctcgat cccctcgcga gttggttcag
     1921 ctgctgcctg aggctggacg acctcgcgga gttctaccgg cagtgcaaat ccgtcggcat
     1981 ccaggaaacc agcagcggct atccgcgcat ccatgccccc gaactgcagg agtggggagg
     2041 cacgatggcc gctttggtcg acccggacgg gacgctcctg cgcctgatac agaacgaatt
     2101 gcttgcaggc atctcatgat cagtactgac aataaaaaga ttcttgtttt caagaacttg
     2161 tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt tagcgtgatt
     2221 tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc gcagaaagta
     2281 atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa
     2341 cgccgccact agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt
     2401 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2461 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2521 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2581 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2641 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2701 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2761 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2821 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2881 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2941 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3001 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3061 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3121 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3181 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3241 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3301 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3361 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3421 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3481 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3541 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3601 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3661 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3721 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3781 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3841 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3901 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3961 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4021 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4081 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4141 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4201 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4261 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4321 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4381 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4441 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4501 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4561 gcacatttcc ccgaaaagtg ccac
//
