LOCUS       sequence-415566-        7900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses NES-TvmvC-43LK-iLID-TvmvN-TVMVseq(P1'S)-mCherry component 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7900)
  AUTHORS   Cui M, Lee S, Ban SH, Ryu JR, Shen M, Yang SH, Kim JY, Choi SK, Han 
            J, Kim Y, Han K, Lee D, Sun W, Kwon HB, Lee D
  TITLE     A single-component, light-assisted uncaging switch for 
            endoproteolytic release.
  JOURNAL   Nat Chem Biol. 2024 Mar;20(3):353-364. doi: 
            10.1038/s41589-023-01480-6. Epub 2023 Nov 16.
  PUBMED    37973890
REFERENCE   2  (bases 1 to 7900)
  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/s41589-023-01480-6"; journalName: "Nat Chem Biol"; date: 
            "2024-03"; volume: "20"; issue: "3"; pages: "353-364"
COMMENT     Sequence Label: pCMV-NES-TvmvC-43LK-iLID-TvmvN-TVMVseq(P1'S)-m...
FEATURES             Location/Qualifiers
     source          1..7900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1888..1914
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1924..1950
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2722..2742
                     /codon_start=1
                     /product="tobacco vein mottling virus (TVMV) NIa protease 
                     recognition and cleavage site
                     "
                     /label=TVMV site
                     /translation="ETVRFQS"
     CDS             2743..3453
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2888..2906)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3156..3175
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3491..3508)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3497..3721
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3767..4195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3854..3873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4064..4085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4204..4224)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4209..4538
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4389..4524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4451..4470
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4605..5399
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4659..4678)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5269..5288
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5573..5694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5610..5629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5664..5683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5743..5759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5743..5759)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5756..5778)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5767..5783
                     /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(5791..5797,5798..5815,5816..5821))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5836..5857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5974..5991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6145..6733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6225..6244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6904..7695,7696..7764))
                     /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     7527..7546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7765..7869)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatga acctggtgga cctgcagaag aagctggagg agctggagct ggacgagcag
      961 cagggaggca agcttaaaag cgtctccagc ctggtgtctg aatcctctca cattgtgcat
     1021 aaagaagaca cttctttctg gcagcactgg atcaccacta aagatggcca gtgtggcagc
     1081 ccacttgttt ccatcattga tggcaacatt ctgggcatcc acagcctgac tcataccacc
     1141 aacggtagca actacttcgt ggaatttccg gaaaaattcg tggcgactta tcttgacgcc
     1201 gcggacggtt ggtgcaaaaa ctggaaattc aacgcggata aaatcagctg gggttccttt
     1261 accctggttg aagatgcgcc ggaagatgac ttcatggcca aaaaaactgt tgccgccatc
     1321 atggacaccg catgtggtgg cggtggctct ggaggtggtg ggtccggagg aggaggtggc
     1381 ggttccggcg gaggtgggtc tggtggaggt ggatctggag gaggtggttc aggtggagga
     1441 ggttctggcg gccgcgaatt tcttgctaca acccttgaaa gaatcgagaa aaatttcgtg
     1501 attactgatc cgagactgcc tgacaaccca attatttttg cgagcgattc cttcctgcaa
     1561 ctgacagaat attctcggga agagatcctg gggcgcaatt gccgttttct gcaaggaccc
     1621 gagacagacc gtgccactgt tcggaaaatc agagatgcta ttgacaacca gactgaagtg
     1681 accgttcagc tgatcaatta taccaagagc ggcaagaagt tctggaacgt gttccacctg
     1741 caaccgatgc gcgattataa gggcgacgtc cagtacttca ttggcgtgca gctggatggc
     1801 accgaacgtc ttcacggcgc cgctgagcgt gaggcggtct gcctcataaa gaaaacggcg
     1861 ttccaaatcg caggatccat gcttcaactt cctcctcttg aacgtcttac tcttatgttg
     1921 cagctaccac cattagagag actaacactg tctaaggctt tgctgaaggg cgtgcgcgat
     1981 tttaatccga tctctgcttg cgtatgcctg ctggaaaact cctcggatgg tcacagtgaa
     2041 cgtctgtttg gcattggttt tggcccgtat atcattgcca accagcacct gtttcgtcgt
     2101 aacaatggcg aactgaccat caaaaccatg cacggtgaat ttaaagtcaa aaactctacc
     2161 cagctgcaaa tgaaacctgt tgaaggccgt gacattatcg ttatcaaaat ggctaaagac
     2221 ttcccgccgt tcccgcagaa actgaaattc cgtcagccga ccatcaaaga tcgtgtgtgc
     2281 atggtgtcca ccaactttca gcaggagttt ctggcaacca cactggaacg gatcgagaaa
     2341 aatttcgtga ttactgatcc gagactgcct gacaacccaa tcatttttgc gagcgattcc
     2401 ttcctgcaac tgacagaata ttctcgggaa gagatcctgg ggcgcaattg ccgttttctg
     2461 caaggacccg agacagaccg tgccactgtt cggaaaatca gagatgctat tgacaaccag
     2521 actgaagtga ccgttcagct gatcaattat accaagagcg gcaagaagtt ctggaacgtg
     2581 ttccacctgc aaccgatgcg cgattataag ggcgacgtcc agtacttcat tggcgtgcag
     2641 ctggatggca ccgaacgtct tcacggcgcc gctgagcgtg aggcggtctg cctgatcaaa
     2701 aagacagcct ttcagattgc tgaaaccgtg cgtttccaga gcatggtgag caagggcgag
     2761 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     2821 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     2881 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     2941 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     3001 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     3061 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     3121 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     3181 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     3241 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     3301 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     3361 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     3421 tccaccggtg gcatggacga gctgtacaag taactcgagt ctagagggcc cgtttaaacc
     3481 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3541 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3601 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3661 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3721 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3781 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3841 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3901 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3961 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4021 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4081 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4141 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4201 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4261 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4321 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4381 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4441 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4501 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4561 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4621 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4681 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4741 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4801 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4861 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4921 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4981 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5041 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5101 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5161 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5221 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5281 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5341 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5401 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5461 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacaccgg
     5521 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5581 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5641 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5701 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5761 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5821 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5881 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5941 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6001 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6061 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6121 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6181 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6241 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6301 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6361 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6421 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6481 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6541 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6601 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6661 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6721 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6841 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6901 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6961 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7021 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7081 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7141 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7201 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7261 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7321 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7381 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7441 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7501 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7621 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7681 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7741 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7801 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7861 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
