LOCUS       sequence-311070-        5326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of KRAB-EL222 in plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5326)
  AUTHORS   Ochoa-Fernandez R, Abel NB, Wieland FG, Schlegel J, Koch LA, Miller 
            JB, Engesser R, Giuriani G, Brandl SM, Timmer J, Weber W, Ott T, 
            Simon R, Zurbriggen MD
  TITLE     Optogenetic control of gene expression in plants in the presence of 
            ambient white light.
  JOURNAL   Nat Methods. 2020 Jul;17(7):717-725. doi: 10.1038/s41592-020-0868-y.
            Epub 2020 Jun 29.
  PUBMED    32601426
REFERENCE   2  (bases 1 to 5326)
  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/s41592-020-0868-y"; journalName: "Nat Methods"; date: 
            "2020-07"; volume: "17"; issue: "7"; pages: "717-725"
COMMENT     Sequence Label: pROF018
FEATURES             Location/Qualifiers
     source          1..5326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..346
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     289..309
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             433..453
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             457..477
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             508..702
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     terminator      1628..1880
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(1931..1949)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1970..1986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1970..1986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1983..2005)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1994..2010
                     /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(2018..2024,2025..2042,2043..2048))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2063..2084
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2201..2218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2372..2960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2452..2471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3131..3922,3923..3991))
                     /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     3754..3773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3992..4096)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4122..4577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4259..4280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4471..4490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4704..4726
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4718..4735
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4719..4735
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4742..4761
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4742..4760
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tgagactttt caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat
       61 ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg
      121 cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc
      181 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
      241 ggattgatgt gatatcacca ctgacgtaag ggatgacgca caatcccact atccttcgca
      301 agacccttcc tctatataag gaagttcatt tcatttggag agaacacggg ggactctagc
      361 gctaccggtt ggctaggtaa gcttggtacc acctgaacga cgcatatgat ctaagctagc
      421 cccaccatgg atccaaaaaa gaagagaaag gtagatccaa aaaagaagag aaaggtaatg
      481 gatgctaagt cactaactgc ctggtcccgg acactggtga ccttcaagga tgtatttgtg
      541 gacttcacca gggaggagtg gaagctgctg gacactgctc agcagatcgt gtacagaaat
      601 gtgatgctgg agaactataa gaacctggtt tccttgggtt atcagcttac taagccagat
      661 gtgatcctcc ggttggagaa gggagaagag ccctggctgg tggagagaga aattcaccaa
      721 gagacccatc ctgattcaga gactgcattt gaaatcaaat catcagtttc cagcaggagc
      781 atttttaaag ataagcaatc ctgtgacatt aaaatggaag gaatggcaag gaatgatctc
      841 tgggaattcg gggcagacga cacacgcgtt gaggtgcaac cgccggcgca gtgggtcctc
      901 gacctgatcg aggccagccc gatcgcatcg gtcgtgtccg atccgcgtct cgccgacaat
      961 ccgctgatcg ccatcaacca ggccttcacc gacctgaccg gctattccga agaagaatgc
     1021 gtcggccgca attgccgatt cctggcaggt tccggcaccg agccgtggct gaccgacaag
     1081 atccgccaag gcgtgcgcga gcacaagccg gtgctggtcg agatcctgaa ctacaagaag
     1141 gacggcacgc cgttccgcaa tgccgtgctc gttgcaccga tctacgatga cgacgacgag
     1201 cttctctatt tcctcggcag ccaggtcgaa gtcgacgacg accagcccaa catgggcatg
     1261 gcgcgccgcg aacgcgccgc ggaaatgctc aagacgctgt cgccgcgcca gctcgaggtt
     1321 acgacgctgg tggcatcggg cttgcgcaac aaggaagtgg cggcccggct cggcctgtcg
     1381 gagaaaaccg tcaagatgca ccgcgggctg gtgatggaaa agctcaacct gaagaccagt
     1441 gccgatctgg tgcgcattgc cgtcgaagcc ggaatctaaa ctagataagt aatgatcata
     1501 atcagccata tcacatctgt agaggtttta cttgctttaa aaaacctccc acacctcccg
     1561 aattccccgg gctcgagaag cttggatcca ccggatctag ataactgatc tcgaggcgaa
     1621 tttccccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     1681 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     1741 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
     1801 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
     1861 gtcatctatg ttactagatc gggaattaat tcgataagct agagatccgt caaggtacca
     1921 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     1981 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2041 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2101 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2161 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2221 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2281 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2341 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2401 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2461 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2521 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2581 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2641 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2701 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2761 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2821 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2881 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2941 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3001 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3061 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3121 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3181 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3241 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3301 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3361 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3421 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3481 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3541 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3601 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3661 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3721 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3781 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3841 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3901 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3961 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4021 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4081 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4141 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4201 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4261 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4321 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4381 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4441 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4501 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4561 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4621 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     4681 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     4741 gtaatacgac tcactatagg gcgaattgga gctccaccgc ggagcggaga attaagggag
     4801 tcaggcctta attaagagct cgcatgcctg caggtcccca gattagcctt ttcaatttca
     4861 gaaagaatgc taacccacag atggttagag aggcttacgc agcaggtctc atcaagacga
     4921 tctacccgag caataatctc caggaaatca aataccttcc caagaaggtt aaagatgcag
     4981 tcaaaagatt caggactaac tgcatcaaga acacagagaa agatatattt ctcaagatca
     5041 gaagtactat tccagtatgg acgattcaag gcttgcttca caaaccaagg caagtaatag
     5101 agattggagt ctctaaaaag gtagttccca ctgaatcaaa ggccatggag tcaaagattc
     5161 aaatagagga cctaacagaa ctcgccgtaa agactggcga acagttcata cagagtctct
     5221 tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac acacttgtct
     5281 actccaaaaa tatcaaagat acagtctcag aagaccaaag ggcaat
//
