LOCUS       sequence-251618-        4215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes nuclear localized PHR domain of CRY2 fused to activating 
            domain of p65 as Type 3 part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4215)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 4215)
  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: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK3_001
FEATURES             Location/Qualifiers
     source          1..4215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             786..806
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             822..842
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             948..2441
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     CDS             2499..2525
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2955..3326
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     terminator      complement(3346..3380)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3451..4110)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4025..4044
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtagtcgg tctcatatgg
      781 ccagccccaa gaagaagcgc aaggtggaag ctagcgcttc cccgaagaaa aagcggaaag
      841 tcgaggcctc cgcatctcca aaaaaaaaaa gcaaggttga agcatctgga tccggtaccg
      901 gaggaagtgg cagctctggc ggcagtggag ggtctggtgg cagcggaatg aagatggaca
      961 aaaagactat agtttggttt agaagggacc taaggattga ggataatcct gcattagcag
     1021 cagctgctca cgaaggatct gtttttcctg tcttcatttg gtgtcctgaa gaagaaggac
     1081 agttttatcc tggaagagct tcaagatggt ggatgaaaca atcacttgct cacttatctc
     1141 aatccttgaa ggctcttgga tctgacctca ctttaatcaa aacccacaac acgatttcag
     1201 cgatcttgga ttgtatccgc gttaccggtg ctacaaaagt cgtctttaac cacctctatg
     1261 atcctgtttc gttagttcgg gaccataccg taaaggagaa gctggtggaa cgtgggatct
     1321 ctgtgcaaag ctacaatgga gatctattgt atgaaccgtg ggagatatac tgcgaaaagg
     1381 gcaaaccttt tacgagtttc aattcttact ggaagaaatg cttagatatg tcgattgaat
     1441 ccgttatgct tcctcctcct tggcggttga tgccaataac tgcagcggct gaagcgattt
     1501 gggcgtgttc gattgaagaa ctagggctgg agaatgaggc cgagaaaccg agcaatgcgt
     1561 tgttaactag agcttggtca ccaggatgga gcaatgctga taagttacta aatgagttca
     1621 tcgagaagca gttgatagat tatgcaaaga acagcaagaa agttgttggg aattctactt
     1681 cactactttc tccgtatctc catttcgggg aaataagcgt cagacacgtt ttccagtgtg
     1741 cccggatgaa acaaattata tgggcaagag ataagaacag tgaaggagaa gaaagtgcag
     1801 atctttttct taggggaatc ggtttaagag agtattctcg gtatatatgt ttcaacttcc
     1861 cgtttactca cgagcaatcg ttgttgagtc atcttcggtt tttcccttgg gatgctgatg
     1921 ttgataagtt caaggcctgg agacaaggca ggaccggtta tccgttggtg gatgccggaa
     1981 tgagagagct ttgggctacc ggatggatgc ataacagaat aagagtgatt gtttcaagct
     2041 ttgctgtgaa gtttcttctc cttccatgga aatggggaat gaagtatttc tgggatacac
     2101 ttttggatgc tgatttggaa tgtgacatcc ttggctggca gtatatctct gggagtatcc
     2161 ccgatggcca cgagcttgat cgcttggaca atcccgcgtt acaaggcgcc aaatatgacc
     2221 cagaaggtga gtacataagg caatggcttc ccgagcttgc gagattgcca actgaatgga
     2281 tccatcatcc atgggacgct cctttaaccg tactcaaagc ttctggtgtg gaactcggaa
     2341 caaactatgc gaaacccatt gtagacatcg acacagctcg tgagctacta gctaaagcta
     2401 tttcaagaac ccgtgaagca cagatcatga tcggagcagc agggggttct ggggggagcg
     2461 gcgggggatc tggaagtggc agcggaggcc tcgagcagta cccctacgac gtgcccgatt
     2521 acgccgagtt ccagtacctg ccagatacag acgatcgtca ccggattgag gagaaacgta
     2581 aaaggacata tgaaaccttc aagagcatca tgaagaagag tcctttcagc ggacccaccg
     2641 acccccggcc tccacctcga cgcattgctg tgccttcccg cagctcagct tctgtcccca
     2701 agccagcacc ccagccctat ccctttacgt catccctgag caccatcaac tatgatgagt
     2761 ttcccaccat ggtgtttcct tctgggcaga tcagccaggc ctcggccttg gccccggccc
     2821 ctccccaagt cctgccccag gctccagccc ctgcccctgc tccagccatg gtatcagctc
     2881 tggcccaggc cccagcccct gtcccagtcc tagccccagg ccctcctcag gctgtggccc
     2941 cacctgcccc caagcccacc caggctgggg aaggaacgct gtcagaggcc ctgctgcagc
     3001 tgcagtttga tgatgaagac ctgggggcct tgcttggcaa cagcacagac ccagctgtgt
     3061 tcacagacct ggcatccgtc gacaactccg agtttcagca gctgctgaac cagggcatac
     3121 ctgtggcccc ccacacaact gagcccatgc tgatggagta ccctgaggct ataactcgcc
     3181 tagtgacagg ggcccagagg ccccccgacc cagctcctgc tccactgggg gccccggggc
     3241 tccccaatgg cctcctttca ggagatgaag acttctcctc cattgcggac atggacttct
     3301 cagccctgct gagtcagatc agctccggat cctgagacca gaccaataaa aaacgcccgg
     3361 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc
     3421 aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta
     3481 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     3541 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
     3601 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     3661 ccagggattg gctgaaacga aaaacatatt ctcaataaac cctttaggga aataggccag
     3721 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     3781 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     3841 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg
     3901 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     3961 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     4021 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     4081 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     4141 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     4201 tacgtgcccg atcaa
//
