LOCUS       sequence-214262-        8337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Venus YFP with a C-terminal SMASh(AI) tag fusion in 
            mammalian cells. [AI = asunaprevir inhibited]..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8337)
  AUTHORS   Jacobs CL, Badiee RK, Lin MZ
  TITLE     StaPLs: versatile genetically encoded modules for engineering 
            drug-inducible proteins.
  JOURNAL   Nat Methods. 2018 Jul;15(7):523-526. doi: 10.1038/s41592-018-0041-z.
            Epub 2018 Jul 2.
  PUBMED    29967496
REFERENCE   2  (bases 1 to 8337)
  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"; doi: 
            "10.1038/s41592-018-0041-z"; journalName: "Nat Methods"; date: 
            "2018-07"; volume: "15"; issue: "7"; pages: "523-526"
COMMENT     Sequence Label: pCS6-YFP-SMASh(AI)
FEATURES             Location/Qualifiers
     source          1..8337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      86..540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(173..192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     382..403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(827..895,896..1687)
                     /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(1045..1064)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    1789..1822
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      1900..2488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2389..2408
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2642..2659
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(2733..2837)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        2947..3250
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3251..3454
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3405..3425
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3451..3475
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     3580..3596
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3580..3596
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3607..3625
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3607..3624
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    3626..3650
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3669..3678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3675..3677,3678..3680,3681..4391)
                     /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(3720..3741)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3981..4000)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4328..4349
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4515..4538
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5367..5393
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(7687..7711)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7712..7731)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7713..7731)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7742..7759)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7742..7758)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7751..7773)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    8163..8297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8200..8219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8254..8273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ggtttgcgta ttggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
       61 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
      121 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      181 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      241 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      301 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
      361 ttggagtcca cgttcttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
      421 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
      481 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
      541 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      601 ttcaaatatg tatccgctca tgccaggtct tggactggtg agaacggctt gctcggcagc
      661 ttcgatgtgt gctggaggga gaataaaggt ctaagatgtg cgatagaggg aagtcgcatt
      721 gaattatgtg ctgtgtaggg atcgctggta tcaaatatgt gtgcccaccc ctggcatgag
      781 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      841 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      901 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      961 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1021 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1081 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1141 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1201 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1261 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1321 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1381 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1441 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1501 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1561 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1621 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1681 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1741 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgccat aacttcgtat
     1801 aatgtatgct atacgaagtt atggcatgac caaaatccct taacgtgagt tttcgttcca
     1861 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1921 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1981 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2041 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2101 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2161 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2221 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2281 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2341 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2401 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2461 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2521 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2581 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2641 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     2701 gcgttggccg attcattaat gcagagcttg caattcgcgc tttttcaata ttattgaagc
     2761 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2821 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     2881 attatcatga cattaaccta taaaaatagg cgtattacga ggccctttca cgcattagat
     2941 gcatgtcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     3001 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     3061 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     3121 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     3181 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     3241 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     3301 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     3361 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     3421 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     3481 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
     3541 gactctagcc taggccgcgg gacggataac aatttcacac aggaaacagc tatgaccatt
     3601 aggcctattt aggtgacact atagaacaag tttgtacaaa aaagcaggct ggtaccggtc
     3661 cggaattcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     3721 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     3781 gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg
     3841 tgccctggcc caccctcgtg accaccctgg gctacggcct gcagtgcttc gcccgctacc
     3901 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     3961 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     4021 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     4081 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg
     4141 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     4201 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     4261 tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc
     4321 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     4381 agctgtacaa gcccgggccc atggccagca tgactggtgg acagcaaatg gggtcgacgc
     4441 tctaccagga gttcgatgag atggaagagt gctctcagca cttacccggc gccggcagta
     4501 gtggcgatat catggattac aaggatgacg acgataaggg ctcttccggg acaggctccg
     4561 gatccggcac tagtgcgccc atcacggcgt acgcccagca gacgagaggc ctcctagggt
     4621 gtataatcac cagcctgact ggccgggaca aaaaccaagt ggagggtgag gtccagatca
     4681 tgtcaactgc tacccaaacc ttcctggcaa cgtgcatcaa tggggtatgc tgggcagtct
     4741 accacggggc cggaacgagg accatcgcat cacccaaggg tcctgtcatc cagatgtata
     4801 ccaatgtgga ccaagacctt gtgggctggc ccgctcctca aggttcccgc tcattgacac
     4861 cctgtacctg cggctcctcg gacctttacc tggtcacgag gcacgccgat gtcattcccg
     4921 tgcgccggcg aggtgatggc aggggtagcc tgctttcgcc ccggcccatt tcctacttga
     4981 aaggctcctc ggggggtccg ctgttgtgcc ccgcgggaca cgccgtgggc ctattcaggg
     5041 ccgcggtgtg cacccgtgga gtggctaaag cggtggactt tatccctgtg gagaacctag
     5101 agacaaccat gagatccccg gtgttcacgg acaactcctc tccaccagca gtcaccctga
     5161 cgcacccaat caccaaaatc gataccaaat acatcatgac atgcatgtcg gccgacctgg
     5221 aggtcgtcac gagcacctgg gtgctcgttg gcggcgtcct ggctgctctg gccgcgtatt
     5281 gcctgtcaac aggctgcgtg gtcatagtgg gcaggatcgt cttgtccggg aagccggcaa
     5341 ttatacctga cagggaggtt ctctactatc cctacgatgt gcccgattac gctggcgcgc
     5401 cttcagtagc cagtgacagg acccagcctg aatagagggg ccagcccagg gtccccagcc
     5461 cgcctgccac acccagcctg tggcttttgc caactaggac ttgagctggg gctgactccc
     5521 aaagggatgc cctgtccagc cagacatcta ctcacccact gacctggcct gactcacaac
     5581 tgccacacaa ccatgctaca tggactgaac atcaagaagc ccctttccca tagggctccc
     5641 acctgccgcc tacccctcat ctgtctgccc tggtcctggc ccccagcccc attggcctca
     5701 ccctctacac tctcagacca tcacagaaca cgatctggct tcctcattct gctccagtgt
     5761 ccagggctct ttgggtaatc aagaaaccaa gtgtctgaaa ggcaacaaaa agtaggcacc
     5821 aaaacccagg ggacatccta gggcagatgc taaagcagag atgctgaggg aacctcaact
     5881 tccggggatg cagcccactc ctgcagacac agcagatcca gctggtaccc tagctgccgc
     5941 ccagggcaac cggccagtct tgggcagcat agctcccctc ccaggggtga gctgaagcca
     6001 caaatgcagc tgaagcagca gacctgacat cctggcacct cctggccccc aggtgagccc
     6061 tgcaccgtgt atcttagagt gggggggggg gcgtgcagaa ccaagtccac tttccaggag
     6121 cccttcaagg cttccctagc aaattagttg caagggtgga gtgaagtggc atcagagtgg
     6181 gtgtcattgt agtgggcact ggcacttcac ggtccaagga cggggctgag gtgggggcgg
     6241 ttctggggga atttgctaag ccagctcacc gaggaccctg tgatttctca ccccagcagt
     6301 gattcatacc agtgaagaag agcagagctc agctccgtga ctcagccatg gcaggcggag
     6361 ggtccccgag gggctgagtc ctcacaccca gccgaggcag cggctggagc ctacagagcc
     6421 aggagaatga caccaggtaa gaaagtcttg cctcctatcc tgaccaagcc tcagatccca
     6481 gccactctcc agatttctgg gttatccggg aagtcctgac aggccccagg ggtaccagct
     6541 gggagcaggg taacctggtg tccctcctgg atctagtggt gagcagttct ctcaccatct
     6601 tgctgcttcc tgtgtccttc aggtctcaag gctgctgaga agtctttgct gccatgtctg
     6661 gaaagggcat caccacacca ccagcaccag caccatcctc tcttctcctg aagctgccta
     6721 catgggttcc aagacacttt caaggcagag aaaacaagat tacagagagg aggtgcctgg
     6781 cagggggcag caccccagct cagcccaaga gctgaaggtg aagacaagcc agcatgaaac
     6841 cacgggtctg ccatgatgcc cgccccgctg gccgctcact agctgcctgc cattagcctc
     6901 tgcagcttga gcagggtctg tacccttggt cctcttggca cagagcccag ctcgctgcat
     6961 ggcctttggc tccccgacca gacccttgca ggagccttaa ggcttgggcc cctgcccagc
     7021 ctgatctttc ctgctgtgcc ctgcctgcca ggtcaagccc agtcccagga gaccccaggc
     7081 cttggctcct aggctgttcc atgaacctcc ctgacctgcc tggtgattgc ccagctgaaa
     7141 cctcagccaa gccccagctc caattacctt gtgctggtag ctgcttgtgt ctgcagtctg
     7201 agtaggcctt gttgcggttc ctccatctgc ctggtctatt ggtgttctga gaccaattcc
     7261 actgatgttc tgacagatcc tccaccctgt gcccctgcca gcccccacaa gtttattttt
     7321 gcacaaaagc catgacccat actcatttgg ctggcatagg gtgtgtaggt aggccctggg
     7381 gactagggag accctggaga tctcaagaga gtgtggctat cccctatttt caccaagcct
     7441 tgaatatcca gccaggctgt ctgcccatac catcttacct caaagacaga tatatatcta
     7501 tatatgattt tgttaataaa actatgaaac ttattggagt atgtgccatt tctcgagggt
     7561 gtgggatggg ctgaagcagg acctcagagg acactgctca aggaggtaca ggaacaccca
     7621 ccgcctatgt accccagaaa aacttctaag tctagagtat ccctcgaggg gcccaagctt
     7681 acgcgtaccc agctttcttg tacaaagtgg tccctatagt gagtcgtatt ataagctagg
     7741 cactggccgt cgttttacaa cgtcgtgact gggaaaactg ctagcttggg atctttgtga
     7801 aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct
     7861 ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actagctgca tatgcttgct
     7921 gcttgagagt tttgcttact gagtatgatt tatgaaaata ttatacacag gagctagtga
     7981 ttctaattgt ttgtgtattt tagattcaca gtcccaaggc tcatttcagg cccctcagtc
     8041 ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     8101 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     8161 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     8221 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     8281 cttatcatgt ctggatcgat cctgcattaa tgaatcggcc aacgcgcggg gagaggc
//
