LOCUS       sequence-415570-        8199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses 
            NTOM20-EGFP-pCMV-PDGFR-TevCs-43LK-iLID(139)-NES-NES-TevN-iLID-TEVseq
            --bArr2-TevCs component in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8199)
  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 8199)
  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-NTOM20-EGFP-TevCs-43LK-iLID(139)-NES-NES-TevN-iLID-TEVseq--bArr
            2-TevCs
FEATURES             Location/Qualifiers
     source          1..8199
                     /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"
     regulatory      1013..1022
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1148..1150,1151..1861)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1148..1150,1151..1861)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1190..1211)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1451..1470)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1798..1819
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2744..2770
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2780..2806
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3581..3601
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5206..5225)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5207..5225)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5230..5364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5254..5273)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5308..5327
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5477..5497)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5477..5495)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5516..5532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5516..5532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5529..5551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5540..5556
                     /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(5564..5570,5571..5588,5589..5594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5609..5630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5747..5764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5917..6505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5997..6016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6676..7467,7468..7536))
                     /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     7299..7318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7537..7641)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7667..8122)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8016..8035
                     /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 tggccaccat
     1021 ggtgggccgc aacagcgcga ttgcagctgg agtgtgcggc gcgctgttca ttggctattg
     1081 catttacttt gatcgcaaac gccgcagcga tccgaacttt aaaggcggtg gagggtcctc
     1141 tagaggggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1201 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1261 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1321 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1381 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1441 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1501 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1561 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1621 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1681 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1741 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1801 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1861 gggttccgga agcggagaat tcaagtccat gtccagcatg gtgagcgata ctagctgtac
     1921 cttcccatca tctgacggaa tcttctggaa gcactggatt cagactaagg acggccagtg
     1981 tggcagccca ctggtgagca cacgagacgg attcatcgtg gggattcaca gcgcctccaa
     2041 ctttacaaac accaataact atttcacctc agtgccaaag aactttatgg agctgctgac
     2101 caaccaggag gcccagcagt gggtgagcgg gtggcgcctg aacgccgatt ccgtgctgtg
     2161 gggcgggcac aaggtgttta tgaccgcatg cggtggcggt ggctctggag gtggtgggtc
     2221 cggaggagga ggtggcggtt ccggcggagg tgggtctggt ggaggtggat ctggaggtgg
     2281 tggatcagga ggaggaggtt caggcggccg cgagtttctg gcaaccacac tggaacggat
     2341 cgagaaaaat ttcgtgatta ctgatccgag actgcctgac aacccaatca tttttgcgag
     2401 cgattccttc ctgcaactga cagaatattc tcgggaagag atcctggggc gcaattgccg
     2461 ttttctgcaa ggacccgaga cagaccgtgc cactgttcgg aaaatcagag atgctattga
     2521 caaccagact gaagtgaccg ttcagctgat caattatacc aagagcggca agaagttctg
     2581 gaacgtgttc cacctgcaac cgatgcgcga ttataagggc gacgtccagt acttcattgg
     2641 cgtgcagctg gatggcaccg aacgtcttca tggcgccgct gagcgtgagg cggtctgcct
     2701 gatcaaaaag acagcctttc agattgctgg atccatgctt caacttcctc ctcttgaacg
     2761 tcttactctc atgttgcagc taccaccatt agagagacta acactgatgg gcgagagcct
     2821 gttcaaggga cctagggact acaaccctat cagtagcaca atttgtcacc tgaccaacga
     2881 gagtgatggc cacacaacaa gcctgtacgg catcgggttc ggacccttta tcatcaccaa
     2941 caagcacctg ttcaggcgga ataatggcac tctgctggtg cagagcctgc acggggtgtt
     3001 caaagtgaag aacacaacca ctctgcagca gcacctgatc gatgggcggg atatgatcat
     3061 cattaggatg cccaaggact tccccccttt tcctcagaaa ctgaagttcc gagagcccca
     3121 gagagaggag agaatctgtc tggtgaccac aaactttcag accgagtttc tggcaaccac
     3181 actggaacgg atcgagaaaa atttcgtgat tactgatccg agactgcctg acaacccaat
     3241 catttttgcg agcgattcct tcctgcagct gacagaatat tctcgggaag agatcctggg
     3301 gcgcaattgc cgttttctgc agggacccga gacagaccgt gccactgttc ggaaaatcag
     3361 agatgctatt gacaaccaga ctgaagtgac cgttcagctg atcaattata ccaagagcgg
     3421 caagaagttc tggaacgtgt tccacctgca gccgatgcgc gattataagg gcgacgtcca
     3481 gtacttcatt ggcgtgcagc tggatggcac cgaacgtctt catggcgccg ctgagcgtga
     3541 ggcggtctgc ctgatcaaaa agacagcctt tcagattgct gagaacctgt acttccaggg
     3601 caccggtatg ggtgaaaaac ccgggaccag ggtcttcaag aagtcgagcc ctaactgcaa
     3661 gctcaccgtg tacttgggca agcgtgactt tgtggatcac ttggacaaag tggatcctgt
     3721 cgatggtgtg gtgcttgtgg atcctgacta cttgaaggac cggaaagtgt ttgtgaccct
     3781 cacctgtgcc ttccgctatg gccgagaaga cctggatgta ctgggcctgt ctttccgcaa
     3841 agatctgttc atcgccacct accaggcctt cccccccatg cccaacccac ctcggccccc
     3901 cacccgccta caggaccgac tgctgaagaa gttgggccag cacgcccacc cctttttttt
     3961 cacaataccc cagaatttgc cttgctccgt cacactgcag ccaggaccgg aggacacagg
     4021 gaaggcctgt ggagtagact ttgagattcg agccttctgt gccaaatcta tagaagaaaa
     4081 aagccacaaa aggaactccg tgcggcttat catcagaaag gtacagtttg ctcctgagac
     4141 acccggcccc cagccatcag ctgaaaccac acgccacttc ctcatgtctg accggaggtc
     4201 cctgcaccta gaggcttccc tggacaaaga gctgtactac catggggaac ccctcaatgt
     4261 caacgtccac gtcaccaaca attctgccaa aaccgtcaag aagatcagag tgtctgtgag
     4321 acagtatgcc gacatttgcc tcttcagcac cgcgcagtac aagtgtcctg tggctcagct
     4381 tgaacaagat gaccaggtgt ctcccagttc cacattctgc aaggtgtaca ccataacccc
     4441 gctgctcagt gacaaccgag agaagcgtgg ccttgccctt gatgggcaac tcaagcacga
     4501 agacaccaac ctggcttcca gcaccattgt gaaggaggga gccaacaagg aggtgctggg
     4561 aatcctagta tcctacaggg tcaaggtgaa gctggtggtg tctcgaggcg gggatgtctc
     4621 cgtggagcta cctttcgtcc taatgcaccc caagccccac gaccacatca cccttccccg
     4681 accccagtca gccccccggg aaatagacat ccctgtggat accaacctca ttgaattcga
     4741 taccaactat gccacagacg acgacatcgt gtttgaggac tttgcgaggc ttcggctgaa
     4801 ggggatgaag gatgacgact gtgatgacca gttctgcctg agctcactcg aaggtggcgg
     4861 tggctctgga ggtggtgggt ccggaggagg cggcatcgag atgtccagca tggtgagcga
     4921 tactagctgt accttcccat catctgacgg aatcttctgg aagcactgga ttcagactaa
     4981 ggacggccag tgtggcagcc cactggtgag cacacgagac ggattcatcg tggggattca
     5041 cagcgcctcc aactttacaa acaccaataa ctatttcacc tcagtgccaa agaactttat
     5101 ggagctgctg accaaccagg aggcccagca gtgggtgagc gggtggcgcc tgaacgccga
     5161 ttccgtgctg tggggcgggc acaaggtgtt tatgtaggct agcctcccta tagtgagtcg
     5221 tattacgtag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     5281 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     5341 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     5401 cagggggagg tgtgggaggt tttttaattc gcgtccgcgg cgccaatgca ttgggcccgg
     5461 tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     5521 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga
     5581 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5641 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5701 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5761 cgctgcgctc ggtcgttcgg ctgcgggagc ggtatcagct cactcaaagg cggtaatacg
     5821 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6661 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6721 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6781 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6841 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6901 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6961 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7021 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7081 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7141 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7201 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7261 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7321 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7381 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7441 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7501 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7561 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7621 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     7681 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     7741 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     7801 gttccggttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     7861 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     7921 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     7981 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     8041 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     8101 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     8161 cgatcggtgc gggcctcttc gctattacgc cagtcgacc
//
