LOCUS       sequence-371336-        5424 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 5424)
  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 5424)
  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-URA
FEATURES             Location/Qualifiers
     source          1..5424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(40..468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(150..171)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     362..381
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     595..617
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     609..626
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     610..626
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     636..655
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        636..654
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     679..695
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     680..696
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             745..834
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             865..1755
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     promoter        1813..2156
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2975..3172
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3246..3266)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3246..3264)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3285..3301)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3285..3301)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3298..3320)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3309..3325
                     /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(3333..3339,3340..3357,3358..3363))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3378..3399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3516..3533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3687..4275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3767..3786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4446..5237,5238..5306))
                     /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     5069..5088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5307..5411)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
       61 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
      121 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
      181 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
      241 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
      301 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
      361 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
      421 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
      481 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
      541 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      601 cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
      661 tgggtaccgg gccccccctc gaggtcgacg gtatcgataa gcttgatatc gggtcgacgg
      721 atccccgggt taattaacat cttttaccca tacgatgttc ctgactatgc gggctatccc
      781 tatgacgtcc cggactatgc aggatcctat ccatatgacg ttccagatta cgctgctcag
      841 tgcggttcag gtggatctgg ttctatggca gaaatcggta ctggctttcc attcgacccg
      901 cattatgtgg aagtcctggg cgagagaatg cattacgttg acgtgggtcc gagagatgga
      961 actccggtcc tttttctgca cgggaatcct acaagctctt atgtttggcg caatatcatc
     1021 cctcatgtag ctccgacgca tcgctgtatt gcgccggacc tgattggtat gggaaaatct
     1081 gataaaccag acctgggtta ctttttcgat gatcatgtgc gtttcatgga tgccttcatt
     1141 gaggcattag ggcttgaaga agtcgtcttg gtgattcatg attggggctc agctctggga
     1201 tttcactggg ctaaaagaaa tcctgaacgc gtaaaaggca tcgcgtttat ggagttcatt
     1261 cgtccaattc cgacttggga tgaatggcct gagttcgcga gagaaacatt tcaagcattt
     1321 cgcacgaccg atgtaggccg gaagttaatc atcgatcaga atgtctttat cgaagggaca
     1381 ttgccgatgg gagtcgttcg tccgttaaca gaagtcgaaa tggatcacta tagagaacct
     1441 tttcttaatc ctgtggacag agagccgctg tggcggtttc cgaacgaact gccgattgca
     1501 ggcgagcctg ctaacattgt agcgctggtt gaagagtata tggattggct tcatcagtct
     1561 ccagttccga agttattgtt ttggggtacg cctggcgtgc ttattccacc ggccgaagcg
     1621 gcacgtttgg caaaaagcct gccaaattgc aaagccgttg acattggccc tggacttaac
     1681 ttgcttcaag aggataaccc ggacttaatc gggagcgaaa ttgcccggtg gctttctacc
     1741 ttagaaatca gcggctaata tataactgtc tagaaataaa gagtatcatc tttcaaaggt
     1801 cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc
     1861 tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca
     1921 tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg
     1981 cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg
     2041 cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt
     2101 ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatga
     2161 cagtcaacac taagacctat agtgagagag cagaaactca tgcctcacca gtagcacaac
     2221 gattatttcg attaatggaa ctgaagaaaa ccaatttatg tgcatcaatt gatgttgata
     2281 ccactaagga attccttgaa ttaattgata aattgggtcc ttatgtatgc ttaatcaaga
     2341 ctcatattga tataatcaat gatttttcct atgaatccac tattgaacca ttattagaac
     2401 tttcacgtaa acatcaattt atgatttttg aagatagaaa atttgctgat attggtaata
     2461 ccgtgaagaa acaatatatt ggtggagttt ataaaattag tagttgggca gatattacta
     2521 atgctcatgg tgtcactggg aatggagtag ttgaaggatt aaaacaggga gctaaagaaa
     2581 ccaccaccaa ccaagagcca agagggttat tgatgttagc tgaattatca tcagtgggat
     2641 cattagcata tggagaatat tctcaaaaaa ctgttgaaat tgctaaatcc gataaggaat
     2701 ttgttattgg atttattgcc caacgtgata tgggtggaca agaagaagga tttgattggc
     2761 ttattatgac acctggagtt ggattagatg ataaaggtga tggattagga caacaatata
     2821 gaactgttga tgaagttgtt agcactggaa ctgatattat cattgttggt agaggattgt
     2881 ttggtaaagg aagagatcca gatattgaag gtaaaaggta tagagatgct ggttggaatg
     2941 cttatttgaa aaagactggc caattataat cagtactgac aataaaaaga ttcttgtttt
     3001 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt
     3061 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc
     3121 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat
     3181 tcgatactaa cgccgccact agttctagag cggccgccac cgcggtggag ctccagcttt
     3241 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     3301 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3361 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3421 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3481 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3541 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3601 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3661 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3721 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3781 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3841 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3901 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3961 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4021 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4081 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4141 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4201 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4261 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4321 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4381 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4441 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4501 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4561 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4621 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4681 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4741 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4801 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4861 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4921 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4981 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5041 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5101 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5161 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5221 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5281 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5341 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5401 ggggttccgc gcacatttcc ccga
//
