LOCUS       sequence-316941-        6212 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SMASh tag fused to YFP at N terminus in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6212)
  AUTHORS   Chung HK, Jacobs CL, Huo Y, Yang J, Krumm SA, Plemper RK, Tsien RY, 
            Lin MZ
  TITLE     Tunable and reversible drug control of protein production via a 
            self-excising degron.
  JOURNAL   Nat Chem Biol. 2015 Jul 27. doi: 10.1038/nchembio.1869.
  PUBMED    26214256
REFERENCE   2  (bases 1 to 6212)
  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: "Nat Chem 
            Biol. 2015 Jul 27. doi: 10.1038/nchembio.1869."
COMMENT     Sequence Label: pCS6-SMASh-YFP
FEATURES             Location/Qualifiers
     source          1..6212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     634..650
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     634..650
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        661..679
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     661..678
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    680..704
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             942..965
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(1860..1862,1863..1865,1866..2579)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1905..1926)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2166..2185)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2513..2534
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2616..2640)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2641..2660)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2642..2660)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2671..2688)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2671..2687)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2680..2702)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    3092..3226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3129..3148)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3183..3202
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3352..3806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3439..3458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3648..3669
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(4093..4161,4162..4953)
                     /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     complement(4311..4330)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    5055..5088
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      5166..5754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5655..5674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5908..5925
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(5999..6103)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agcctaggcc gcgggacgga taacaatttc acacaggaaa cagctatgac cattaggcct
      661 atttaggtga cactatagaa caagtttgta caaaaaagca ggctggtacc ggtccggaat
      721 tcggtgcaga gctcaagcga gttctcccgc agccgcagtc tctgggcctc tctagcttca
      781 gcggcgacga gcctgccaca ctcgctaagc tcctccggca ccgcacacct gccactgccg
      841 ctgcagccgc cggctctgct cccttccggc ttctgcctca gaggagttct tagcctgttc
      901 ggagccgcag caccgacgac cagatggagc ccgggcccat ggattacaag gatgacgacg
      961 ataagggctc ttccgggaca ggctccggat ccggcactag tgcgcccatc acggcgtacg
     1021 cccagcagac gagaggcctc ctagggtgta taatcaccag cctgactggc cgggacaaaa
     1081 accaagtgga gggtgaggtc cagatcgtgt caactgctac ccaaaccttc ctggcaacgt
     1141 gcatcaatgg ggtatgctgg gcagtctacc acggggccgg aacgaggacc atcgcatcac
     1201 ccaagggtcc tgtcatccag atgtatacca atgtggacca agaccttgtg ggctggcccg
     1261 ctcctcaagg ttcccgctca ttgacaccct gtacctgcgg ctcctcggac ctttacctgg
     1321 tcacgaggca cgccgatgtc attcccgtgc gccggcgagg tgatagcagg ggtagcctgc
     1381 tttcgccccg gcccatttcc tacttgaaag gctcctcggg gggtccgctg ttgtgccccg
     1441 cgggacacgc cgtgggccta ttcagggccg cggtgtgcac ccgtggagtg gctaaagcgg
     1501 tggactttat ccctgtggag aacctagaga caaccatgag atccccggtg ttcacggaca
     1561 actcctctcc accagcagtc accctgacgc acccaatcac caaaatcgat accaaataca
     1621 tcatgacatg catgtcggcc gacctggagg tcgtcacgag cacctgggtg ctcgttggcg
     1681 gcgtcctggc tgctctggcc gcgtattgcc tgtcaacagg ctgcgtggtc atagtgggca
     1741 ggatcgtctt gtccgggaag ccggcaggca gtagcggaag cagtattata cctgacaggg
     1801 aggttctcta ccaggagttc gaagatgtcg tgccatgctc aatgggctcg ggcgcgccta
     1861 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1921 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1981 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2041 tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     2101 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2161 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2221 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2281 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     2341 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     2401 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2461 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2521 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgat
     2581 ctagagtatc cctcgagggg cccaagctta cgcgtaccca gctttcttgt acaaagtggt
     2641 ccctatagtg agtcgtatta taagctaggc actggccgtc gttttacaac gtcgtgactg
     2701 ggaaaactgc tagcttggga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt
     2761 ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata
     2821 atgtgttaaa ctagctgcat atgcttgctg cttgagagtt ttgcttactg agtatgattt
     2881 atgaaaatat tatacacagg agctagtgat tctaattgtt tgtgtatttt agattcacag
     2941 tcccaaggct catttcaggc ccctcagtcc tcacagtctg ttcatgatca taatcagcca
     3001 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3061 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcgatc ctgcattaat
     3241 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg ctggcgtaat agcgaagagg
     3301 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     3361 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3421 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3481 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3541 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3601 gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt
     3661 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3721 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3781 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     3841 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgcc aggtcttgga
     3901 ctggtgagaa cggcttgctc ggcagcttcg atgtgtgctg gagggagaat aaaggtctaa
     3961 gatgtgcgat agagggaagt cgcattgaat tatgtgctgt gtagggatcg ctggtatcaa
     4021 atatgtgtgc ccacccctgg catgagacaa taaccctgat aaatgcttca ataatattga
     4081 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4141 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4201 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4261 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4321 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4381 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4441 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4501 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4561 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4621 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4681 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4741 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4801 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4861 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4921 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4981 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5041 gataatctca tgccataact tcgtataatg tatgctatac gaagttatgg catgaccaaa
     5101 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5161 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5221 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5281 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5341 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5401 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5461 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5521 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5581 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5641 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5701 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5761 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5821 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5881 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5941 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag agcttgcaat
     6001 tcgcgctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6061 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6121 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6181 ttacgaggcc ctttcacgca ttagatgcat gt
//
