LOCUS       sequence-247308-        8256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SMASh tag fused to YFP at C terminus in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8256)
  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 8256)
  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-YFP-SMASh
FEATURES             Location/Qualifiers
     source          1..8256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        312..515
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     466..486
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     512..536
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     641..657
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     641..657
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        668..686
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     668..685
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    687..711
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      730..739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(736..738,739..741,742..1452)
                     /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(781..802)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1042..1061)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1389..1410
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1519..1542
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(4667..4691)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4692..4711)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4693..4711)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4722..4739)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4722..4738)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4731..4753)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    5143..5277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5180..5199)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5234..5253
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5403..5857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5490..5509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5699..5720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(6144..6212,6213..7004)
                     /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(6362..6381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    7106..7139
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      7217..7805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7706..7725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7959..7976
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(8050..8154)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgcatgtcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
       61 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      121 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      181 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      241 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      301 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      361 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      421 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      481 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct
      541 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
      601 ggactctagc ctaggccgcg ggacggataa caatttcaca caggaaacag ctatgaccat
      661 taggcctatt taggtgacac tatagaacaa gtttgtacaa aaaagcaggc tggtaccggt
      721 ccggaattcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      781 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      841 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
      901 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
      961 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1021 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1081 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1141 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1201 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1261 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1321 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     1381 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1441 gagctgtaca agcccgggga tgagatggaa gagtgctctc agcacttacc cggcgccggc
     1501 agtagtggcg atatcatgga ttacaaggat gacgacgata agggctcttc cgggacaggc
     1561 tccggatccg gcactagtgc gcccatcacg gcgtacgccc agcagacgag aggcctccta
     1621 gggtgtataa tcaccagcct gactggccgg gacaaaaacc aagtggaggg tgaggtccag
     1681 atcgtgtcaa ctgctaccca aaccttcctg gcaacgtgca tcaatggggt atgctgggca
     1741 gtctaccacg gggccggaac gaggaccatc gcatcaccca agggtcctgt catccagatg
     1801 tataccaatg tggaccaaga ccttgtgggc tggcccgctc ctcaaggttc ccgctcattg
     1861 acaccctgta cctgcggctc ctcggacctt tacctggtca cgaggcacgc cgatgtcatt
     1921 cccgtgcgcc ggcgaggtga tagcaggggt agcctgcttt cgccccggcc catttcctac
     1981 ttgaaaggct cctctggggg tccgctgttg tgccccgcgg gacacgccgt gggcctattc
     2041 agggccgcgg tgtgcacccg tggagtggct aaagcggtgg actttatccc tgtggagaac
     2101 ctagagacaa ccatgagatc cccggtgttc acggacaact cctctccacc agcagtcacc
     2161 ctgacgcacc caatcaccaa aatcgatacc aaatacatca tgacatgcat gtcggccgac
     2221 ctggaggtcg tcacgagcac ctgggtgctc gttggcggcg tcctggctgc tctggccgcg
     2281 tattgcctgt caacaggctg cgtggtcata gtgggcagga tcgtcttgtc cgggaagccg
     2341 gcaattatac ctgacaggga ggttctctac tgaggcgcgc cttcagtagc cagtgacagg
     2401 acccagcctg aatagagggg ccagcccagg gtccccagcc cgcctgccac acccagcctg
     2461 tggcttttgc caactaggac ttgagctggg gctgactccc aaagggatgc cctgtccagc
     2521 cagacatcta ctcacccact gacctggcct gactcacaac tgccacacaa ccatgctaca
     2581 tggactgaac atcaagaagc ccctttccca tagggctccc acctgccgcc tacccctcat
     2641 ctgtctgccc tggtcctggc ccccagcccc attggcctca ccctctacac tctcagacca
     2701 tcacagaaca cgatctggct tcctcattct gctccagtgt ccagggctct ttgggtaatc
     2761 aagaaaccaa gtgtctgaaa ggcaacaaaa agtaggcacc aaaacccagg ggacatccta
     2821 gggcagatgc taaagcagag atgctgaggg aacctcaact tccggggatg cagcccactc
     2881 ctgcagacac agcagatcca gctggtaccc tagctgccgc ccagggcaac cggccagtct
     2941 tgggcagcat agctcccctc ccaggggtga gctgaagcca caaatgcagc tgaagcagca
     3001 gacctgacat cctggcacct cctggccccc aggtgagccc tgcaccgtgt atcttagagt
     3061 gggggggggg gcgtgcagaa ccaagtccac tttccaggag cccttcaagg cttccctagc
     3121 aaattagttg caagggtgga gtgaagtggc atcagagtgg gtgtcattgt agtgggcact
     3181 ggcacttcac ggtccaagga cggggctgag gtgggggcgg ttctggggga atttgctaag
     3241 ccagctcacc gaggaccctg tgatttctca ccccagcagt gattcatacc agtgaagaag
     3301 agcagagctc agctccgtga ctcagccatg gcaggcggag ggtccccgag gggctgagtc
     3361 ctcacaccca gccgaggcag cggctggagc ctacagagcc aggagaatga caccaggtaa
     3421 gaaagtcttg cctcctatcc tgaccaagcc tcagatccca gccactctcc agatttctgg
     3481 gttatccggg aagtcctgac aggccccagg ggtaccagct gggagcaggg taacctggtg
     3541 tccctcctgg atctagtggt gagcagttct ctcaccatct tgctgcttcc tgtgtccttc
     3601 aggtctcaag gctgctgaga agtctttgct gccatgtctg gaaagggcat caccacacca
     3661 ccagcaccag caccatcctc tcttctcctg aagctgccta catgggttcc aagacacttt
     3721 caaggcagag aaaacaagat tacagagagg aggtgcctgg cagggggcag caccccagct
     3781 cagcccaaga gctgaaggtg aagacaagcc agcatgaaac cacgggtctg ccatgatgcc
     3841 cgccccgctg gccgctcact agctgcctgc cattagcctc tgcagcttga gcagggtctg
     3901 tacccttggt cctcttggca cagagcccag ctcgctgcat ggcctttggc tccccgacca
     3961 gacccttgca ggagccttaa ggcttgggcc cctgcccagc ctgatctttc ctgctgtgcc
     4021 ctgcctgcca ggtcaagccc agtcccagga gaccccaggc cttggctcct aggctgttcc
     4081 atgaacctcc ctgacctgcc tggtgattgc ccagctgaaa cctcagccaa gccccagctc
     4141 caattacctt gtgctggtag ctgcttgtgt ctgcagtctg agtaggcctt gttgcggttc
     4201 ctccatctgc ctggtctatt ggtgttctga gaccaattcc actgatgttc tgacagatcc
     4261 tccaccctgt gcccctgcca gcccccacaa gtttattttt gcacaaaagc catgacccat
     4321 actcatttgg ctggcatagg gtgtgtaggt aggccctggg gactagggag accctggaga
     4381 tctcaagaga gtgtggctat cccctatttt caccaagcct tgaatatcca gccaggctgt
     4441 ctgcccatac catcttacct caaagacaga tatatatcta tatatgattt tgttaataaa
     4501 actatgaaac ttattggagt atgtgccatt tctcgagggt gtgggatggg ctgaagcagg
     4561 acctcagagg acactgctca aggaggtaca ggaacaccca ccgcctatgt accccagaaa
     4621 aacttctaag tctagagtat ccctcgaggg gcccaagctt acgcgtaccc agctttcttg
     4681 tacaaagtgg tccctatagt gagtcgtatt ataagctagg cactggccgt cgttttacaa
     4741 cgtcgtgact gggaaaactg ctagcttggg atctttgtga aggaacctta cttctgtggt
     4801 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
     4861 ttaagtgtat aatgtgttaa actagctgca tatgcttgct gcttgagagt tttgcttact
     4921 gagtatgatt tatgaaaata ttatacacag gagctagtga ttctaattgt ttgtgtattt
     4981 tagattcaca gtcccaaggc tcatttcagg cccctcagtc ctcacagtct gttcatgatc
     5041 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     5101 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     5161 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5221 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcgat
     5281 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg gctggcgtaa
     5341 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5401 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5461 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5521 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5581 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5641 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tcttaatagt
     5701 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5761 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5821 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     5881 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatgc
     5941 caggtcttgg actggtgaga acggcttgct cggcagcttc gatgtgtgct ggagggagaa
     6001 taaaggtcta agatgtgcga tagagggaag tcgcattgaa ttatgtgctg tgtagggatc
     6061 gctggtatca aatatgtgtg cccacccctg gcatgagaca ataaccctga taaatgcttc
     6121 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6181 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6241 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6301 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6361 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6421 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6481 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6541 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6601 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6661 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6721 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6781 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7021 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7081 agatcctttt tgataatctc atgccataac ttcgtataat gtatgctata cgaagttatg
     7141 gcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7201 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7261 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7321 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7381 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7441 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7501 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7561 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7621 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7681 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7741 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7801 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7861 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7921 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7981 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     8041 gagcttgcaa ttcgcgcttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8101 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8161 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     8221 aaataggcgt attacgaggc cctttcacgc attaga
//
