LOCUS       sequence-136499-        5574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes CelTag for the tagging and high affinity purification of 
            endogenous yeast proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5574)
  AUTHORS   Carrick BH, Hao L, Smaldino PJ, Engelke DR
  TITLE     A Novel Recombinant DNA System for High Efficiency Affinity 
            Purification of Proteins in Saccharomyces cerevisiae.
  JOURNAL   G3 (Bethesda). 2015 Dec 29. pii: g3.115.025106. doi: 
            10.1534/g3.115.025106.
  PUBMED    26715090
REFERENCE   2  (bases 1 to 5574)
  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). 2015 Dec 29. pii: g3.115.025106. doi: 
            10.1534/g3.115.025106."
COMMENT     Sequence Label: CelTag Plasmid
FEATURES             Location/Qualifiers
     source          1..5574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      33..42
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      37..46
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             94..591
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
     CDS             619..639
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      1126..1313
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1181..1205)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1388..1731
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2343..2540
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2593..2609)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2594..2610)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2699..2717)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2700..2717)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2735..2751)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2735..2751)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2748..2770)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2759..2775
                     /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(2783..2789,2790..2807,2808..2813))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2828..2849
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2966..2983)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3137..3725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3217..3236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3896..4687,4688..4756))
                     /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     4519..4538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4757..4861)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4939..5394)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5076..5097)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5288..5307
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5520..5542
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5534..5551
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5535..5551
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5558..5574,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5558..5574,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 taatcgatac atatgcccgg gttaattaac ggtgaacaaa agctaatctc cgaggaagac
      121 ttgaacggtg aacaaaaatt aatctcagaa gaagacttga acggactcga cggtgaacaa
      181 aagttgattt ctgaagaaga tttgaacggt gaacaaaagc taatctccga ggaagacttg
      241 aacggtgaac aaaaattaat ctcagaagaa gacttgaacg gactcgacgg tgaacaaaag
      301 ttgatttctg aagaagattt gaacggtgaa caaaagctaa tctccgagga agacttgaac
      361 ggtgaacaaa aattaatctc agaagaagac ttgaacggac tcgacggtga acaaaagttg
      421 atttctgaag aagatttgaa cggtgaacaa aagctaatct ccgaggaaga cttgaacggt
      481 gaacaaaaat taatctcaga agaagacttg aacggactcg acggtgaaca aaagttgatt
      541 tctgaagaag atttgaacgg tgaacaaaag ctaatctccg aggaagactt gaacggtgaa
      601 caaaaattaa tcaatcacga gaacctctac ttccaaagcg tatcaggcaa tttgaaggtt
      661 gaattctaca acagcaatcc ttcagatact actaactcaa tcaatcctca gttcaaggtt
      721 actaataccg gaagcagtgc aattgatttg tccaaactca cattgagata ttattataca
      781 gtagacggac agaaagatca gaccttctgg tgtgaccatg ctgcaataat cggcagtaac
      841 ggcagctaca acggaattac ttcaaatgta aaaggaacat ttgtaaaaat gagttcctca
      901 acaaataacg cagacaccta ccttgaaata agctttacag gcggaactct tgaaccgggt
      961 gcacatgttc agatacaagg tagatttgca aagaatgact ggagtaacta tacacagtca
     1021 aatgactact cattcaagtc tgcttcacag tttgttgaat gggatcaggt aacagcatac
     1081 ttgaacggtg ttcttgtatg gggtaaagaa tagacttcta aataagcgaa tttcttatga
     1141 tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt
     1201 gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg
     1261 ttgctttctc aggtatagta tgaggtcgct cttattgacc acacctctac cggcagatcc
     1321 gctagggata acagggtaat atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc
     1381 ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg
     1441 ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc
     1501 atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac
     1561 ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct
     1621 gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt
     1681 ttaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac catgggtacc
     1741 actcttgacg acacggctta ccggtaccgc accagtgtcc cgggggacgc cgaggccatc
     1801 gaggcactgg atgggtcctt caccaccgac accgtcttac tggatgggtc cttcaccacc
     1861 gacaccgtct tccgcgtcac cgccaccggg gacggcttca ccctgcggga ggtgccggtg
     1921 gacccgcccc tgaccaaggt gttccccgac gacgaatcgg acgacgaatc ggacgacggg
     1981 gaggacggcg acccggactc tcggacgttc gtcgcgtacg gggacgacgg cgacctggcg
     2041 ggcttcgtgg tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga ggacatcgag
     2101 gtcgccccgg agcaccgggg gcacggggtc gggcgcgcgt tgatggggct cgcgacggag
     2161 ttcgcccgcg agcggggtgc cgggcacctc tggctggagg tcaccaacgt caacgcaccg
     2221 gcgatccacg cgtaccggcg gatggggttc accctctgcg gcctggacac cgccctgtac
     2281 gacggcaccg cctcggacgg cgagcaggcg ctctacatga gcatgccctg cccctaatca
     2341 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     2401 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     2461 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     2521 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga
     2581 gctcgaattc atcgataccg tcgacctcga gcgtacgtaa tcactagtga attcgcggcc
     2641 gcctgcaggt cgaccatatg ggagagctcc caacgcgttg gatgcatagc ttgagtattc
     2701 tatagtgtca cctaaatagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2761 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2821 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2881 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2941 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3001 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3061 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3121 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3181 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3241 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3301 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3361 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3421 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3481 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3541 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3601 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3661 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3721 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3781 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3841 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3901 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3961 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4021 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4081 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4141 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4201 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4261 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4321 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4561 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4741 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4801 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4861 gcacatttcc ccgaaaagtg ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg
     4921 agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4981 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     5041 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     5101 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     5161 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     5221 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     5281 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     5341 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc
     5401 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     5461 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     5521 ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tata
//
