LOCUS       sequence-415564-        6653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses 
            NLSx2-TVMVpC-43LK-iLID(139)-NES-NES-TVMVpN-iLID-TVMVseq-Src13-GB1-LR
            P6c component in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6653)
  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 6653)
  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-NLSx2-TVMVpC-43LK-iLID(139)-NES-NES-TVMVpN-iLID-TVMVseq-Src13-G
            B1-LRP6c
FEATURES             Location/Qualifiers
     source          1..6653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     CDS             1021..1041
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1042..1062
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2764..2802
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="GSSKSKPKDPSQR"
     CDS             2824..2985
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     primer_bind     complement(3660..3679)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3661..3679)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3684..3818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3708..3727)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3762..3781
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3931..3951)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3931..3949)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3970..3986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3970..3986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3983..4005)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3994..4010
                     /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(4018..4024,4025..4042,4043..4048))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4063..4084
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4201..4218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4371..4959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4451..4470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5130..5921,5922..5990))
                     /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     5753..5772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5991..6095)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6121..6576)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6470..6489
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgccaccatg
     1021 cctaaaaaga agcgtaaagt cccgaagaag aagaggaagg tgtctggagg actcgagaaa
     1081 agcgtctcca gcctggtgtc tgaatcctct cacattgtgc ataaagaaga cacttctttc
     1141 tggcagcact ggatcaccac taaagatggc cagtgtggca gcccacttgt ttccatcatt
     1201 gatggcaaca ttctgggcat ccacagcctg actcatacca ccaacggtag caactacttc
     1261 gtggaatttc cggaaaaatt cgtggcgact tatcttgacg ccgcggacgg ttggtgcaaa
     1321 aactggaaat tcaacgcgga taaaatcagc tggggttcct ttaccctggt tgaagatgcg
     1381 ccggaagatg acttcatggc caaaaaaact gttgccgcca tcatggacac cgcatgtggt
     1441 ggcggtggct ctggaggtgg tgggtccgga ggaggaggtg gcggttccgg cggaggtggg
     1501 tctggtggag gtggatctgg aggaggtggt tcaggtggag gaggttctgg cggccgcgaa
     1561 tttcttgcta caacccttga aagaatcgag aaaaatttcg tgattactga tccgagactg
     1621 cctgacaacc caatcatttt tgcgagcgat tccttcctgc aactgacaga atattctcgg
     1681 gaagagatcc tggggcgcaa ttgccgtttt ctgcaaggac ccgagacaga ccgtgccact
     1741 gttcggaaaa tcagagatgc tattgacaac cagactgaag tgaccgttca gctgatcaat
     1801 tataccaaga gcggcaagaa gttctggaac gtgttccacc tgcaaccgat gcgcgattat
     1861 aagggcgacg tccagtactt cattggcgtg cagctggatg gcaccgaacg tcttcacggc
     1921 gccgctgagc gtgaggcggt ctgcctgatc aaaaagacag cctttcagat tgcttctaag
     1981 gctttgctga agggcgtgcg cgattttaat ccgatctctg cttgcgtatg cctgctggaa
     2041 aactcctcgg atggtcacag tgaacgtctg tttggcattg gttttggccc gtatatcatt
     2101 gccaaccagc acctgtttcg tcgtaacaat ggcgaactga ccatcaaaac catgcacggt
     2161 gaatttaaag tcaaaaactc tacccagctg caaatgaaac ctgttgaagg ccgtgacatt
     2221 atcgttatca aaatggctaa agacttcccg ccgttcccgc agaaactgaa attccgtcag
     2281 ccgaccatca aagatcgtgt gtgcatggtg tccaccaact ttcagcagga gtttctggca
     2341 accacactgg aacggatcga gaaaaatttc gtgattactg atccgagact gcctgacaac
     2401 ccaatcattt ttgcgagcga ttccttcctg caactgacag aatattctcg ggaagagatc
     2461 ctggggcgca attgccgttt tctgcaagga cccgagacag accgtgccac tgttcggaaa
     2521 atcagagatg ctattgacaa ccagactgaa gtgaccgttc agctgatcaa ttataccaag
     2581 agcggcaaga agttctggaa cgtgttccac ctgcaaccga tgcgcgatta taagggcgac
     2641 gtccagtact tcattggcgt gcagctggat ggcaccgaac gtcttcacgg cgccgctgag
     2701 cgtgaggcgg tctgcctgat caaaaagaca gcctttcaga ttgctgaaac cgtgcgtttc
     2761 cagggaagca gcaagagcaa gcccaaggac cccagccaga ggggtggagg tcggaccgaa
     2821 gagtacaagc ttatcctgaa cggtaaaacc ctgaaaggtg aaaccaccac cgaagctgtt
     2881 gacgctgcta ccgcggaaaa agttttcaaa cagtacgcta acgacaacgg tgttgacggt
     2941 gaatggacct acgacgacgc taccaaaacc ttcacggtaa ccgaaggtga gctcaggatg
     3001 ttgtgtccac gtatgaaggg agatggggaa actatgacta atgactatgt agttcatgga
     3061 ccagcttctg tgcctcttgg ttatgtgcca cacccaagtt ctttgtcagg atctcttcca
     3121 ggaatgagca gaggtaaatc aatgatcagc tccctcagta tcatgggggg aagcagtgga
     3181 cccccctatg accgagccca tgttacagga gcatcatcaa gtagttcttc aagcaccaaa
     3241 ggcacttact tccctgcaat tttgaaccct ccaccatccc cagccacaga gcgatcacat
     3301 tacactatgg aatttggata ttcttcaaac agtccttcca ctcataggtc atacagctac
     3361 aggccatata gctaccggca ctttgcaccc cccaccacac cctgtagcac agatgtttgt
     3421 gacagtgact atgctcctag tcggagaatg acctcagtgg caacagccaa gggctatacc
     3481 agtgacttga actatgattc agaacctgtg cccccacctc ccacaccccg aagccaatac
     3541 ttgtcagcag aggagaacta tgaaagctgc ccaccttctc catacacaga gaggagctat
     3601 tctcatcacc tctacccacc gccaccctct ccctgtacag actcctccta agctagcctc
     3661 cctatagtga gtcgtattac gtagatccag acatgataag atacattgat gagtttggac
     3721 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     3781 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     3841 ttatgtttca ggttcagggg gaggtgtggg aggtttttta attcgcgtcc gcggcgccaa
     3901 tgcattgggc ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     3961 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4021 catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4081 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4141 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4201 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg gagcggtatc agctcactca
     4261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4381 ctccgccccc ctgacgagca tcacaaaaat cgacgatcaa gtcagaggtg gcgaaacccg
     4441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4801 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4921 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5161 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5221 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5281 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5341 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5401 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5461 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5521 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5581 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5641 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5701 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5761 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5821 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5881 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5941 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6001 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6061 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6121 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     6181 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     6241 tagggttgag tgttgttccg gtttggaaca agagtccact attaaagaac gtggactcca
     6301 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     6361 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     6421 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     6481 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     6541 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     6601 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg acc
//
