LOCUS       sequence-361255-        6162 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Single-component photo-inducible (LOV2-based) nucleolus-targeting 
            tool 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6162)
  AUTHORS   Tan P, Hong T, Cai X, Li W, Huang Y, He L, Zhou Y
  TITLE     Optical control of protein delivery and partitioning in the 
            nucleolus.
  JOURNAL   Nucleic Acids Res. 2022 Mar 23. pii: 6553118. doi: 
            10.1093/nar/gkac191.
  PUBMED    35325178
REFERENCE   2  (bases 1 to 6162)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2022 Mar 23. pii: 6553118. doi: 10.1093/nar/gkac191."
COMMENT     Sequence Label: pNUT1
FEATURES             Location/Qualifiers
     source          1..6162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     183..1010
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             183..307
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(345..950)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        1078..1381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1383..1585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1536..1556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1788..1810
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1872..1891
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1933..1952
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1933..1951
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1952..1976
                     /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        1992..2101
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             2122..2139
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2146..2850
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2288..2306)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2556..2575
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3346..3369
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(3459..3478)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3524..3579
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3578..3597)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      3667..3714
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3727..4432
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3815..4187
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3844..4432)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(4441..4474)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4642..5229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4721..4740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5242..6033,6034..6102))
                     /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     5865..5884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tttcctgcat cttttaatca aatcccaaga tgtgtataaa cgcgccggta tgtacaggaa
       61 gaggtttata ctaaactgtt acattgcaaa cgtggtttcg tgtgccaagt gtgaaaaccg
      121 atgtttaatc aaggctctga cgcatttcta caaccacgac tccaagtgtg tgggtgaagt
      181 catgcatctt ttaatcaaat cccaagatgt gtataaacca ccaaactgcc aaaaaatgaa
      241 aactgtcgac aagctctgtc cgtttgctgg caactgcaag ggtctcaatc ctatttgtaa
      301 ttattgaata ataaaacaat tataaatgtc aaatttgttt tttattaacg atacaaacca
      361 aacgcaacaa gaacatttgt agtattatct ataattgaaa acgcgtagtt ataatcgctg
      421 aggtaatatt taaaatcatt ttcaaatgat tcacagttaa tttgcgacaa tataatttta
      481 ttttcacata aactagacgc cttgtcgtct tcttcttcgt attccttctc tttttcattt
      541 ttctcttcat aaaaattaac atagttatta tcgtatccat atatgtatct atcgtataga
      601 gtaaattttt tgttgtcata aatatatatg tcttttttaa tggggtgtat agtaccgctg
      661 cgcatagttt ttctgtaatt tacaacagtg ctattttctg gtagttcttc ggagtgtgtt
      721 gctttaatta ttaaatttat ataatcaatg aatttgggat cgtcggtttt gtacaatatg
      781 ttgccggcat agtacgcagc ttcttctagt tcaattacac cattttttag cagcaccgga
      841 ttaacataac tttccaaaat gttgtacgaa ccgttaaaca aaaacagttc acctcccttt
      901 tctatactat tgtctgcgag cagttgtttg ttgttaaaaa taacagccat tgtaatgaga
      961 cgcacaaact aatatcacaa actggaaatg tctatcaata tatagttgct gatggccggc
     1021 ctattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     1081 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     1141 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     1201 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     1261 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1321 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1381 gctgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
     1441 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1501 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
     1561 gtgggaggtc tatataagca gacgtcgttt agtgaaccgt cagatcacta gatgctttat
     1621 tgcggtagtt tatcacagtt aaattgctaa cgccagtctc gaacttaacg tgcagaagtt
     1681 ggtcgtgagg cactgggcag gtaagtatcg ggccctttgt gcggggggag cggctcgggg
     1741 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     1801 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
     1861 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgga
     1921 tcggaccgaa attaatacga ctcactatag gggaattgtg agcggataac aattccccgg
     1981 agttaatccg ggacctttaa ttcaacccaa cacaatatat tatagttaaa taagaattat
     2041 tatcaaatca tttgtatatt aattaaaata ctatactgta aattacattt tatttacaat
     2101 caaaggagat ataccatggc acaccatcac caccatcacg gatctgtgag caagggcgag
     2161 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     2221 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     2281 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     2341 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     2401 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     2461 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     2521 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagactatg
     2581 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     2641 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     2701 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     2761 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     2821 tccaccggcg gcatggacga gctgtacaag ggatccttgg ctactacact tgaacgtatt
     2881 gagaagaact ttgtcattac tgacccaaga ttgccagata atcccattat attcgcgtcc
     2941 gatagtttct tgcagttgac agaatatagc cgtgaagaaa ttttgggaag aaactgcagg
     3001 tttctacaag gtcctgaaac tgatcgcgcg acagtgagaa aaattagaga tgccatagat
     3061 aaccaaacag aggtcactgt tcagctgatt aattatacaa agagtggtaa aaagttctgg
     3121 aacctctttc acttgcagcc tatgcgagat cagaagggag atgtccagta ctttattggg
     3181 gttcagttgg atggaactga gcatgtccga gatgctgccg agagagaggg agtcatgctg
     3241 attaagaaaa ctgcagaaaa tattgatgag gcggcaaaag aacttaagct tcgacgaaac
     3301 cgccggagac gctggagaga gcggcaacgg cagatctagc tcgagcacca ccatcaccat
     3361 caccatcact aagtgattaa cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     3421 tggccaatgc cctggctcac aaataccact gagatcgatc tttttccctc tgccaaaaat
     3481 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     3541 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     3601 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     3661 atgtaactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     3721 gcatgcggag gaaattctcc ttgaagtttc cctggtgttc aaagtaaagg agtttgcacc
     3781 agacgcacct ctgttcactg gtccggcgta ttaaaacacg atacattgtt attagtacat
     3841 ttattaagcg ctagattctg tgcgttgttg atttacagac aattgttgta cgtattttaa
     3901 taattcatta aatttataat ctttagggtg gtatgttaga gcgaaaatca aatgattttc
     3961 agcgtcttta tatctgaatt taaatattaa atcctcaata gatttgtaaa ataggtttcg
     4021 attagtttca aacaagggtt gtttttccga accgatggct ggactatcta atggattttc
     4081 gctcaacgcc acaaaacttg ccaaatcttg tagcagcaat ctagctttgt cgatattcgt
     4141 ttgtgttttg ttttgtaata aaggttcgac gtcgttcaaa atattatgcg cttttgtatt
     4201 tctttcatca ctgtcgttag tgtacaattg actcgacgta aacacgttaa atagagcttg
     4261 gacatattta acatcgggcg tgttagcttt attaggccga ttatcgtcgt cgtcccaacc
     4321 ctcgtcgtta gaagttgctt ccgaagacga ttttgccata gccacacgac gcctattaat
     4381 tgtgtcggct aacacgtccg cgatcaaatt tgtagttgag ctttttggaa ttgcgatcgc
     4441 ataacttcgt atagcataca ttatacgaag ttataagctc ggaacgctgc gctcggtcgt
     4501 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4561 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc ctgacgagca tcacaaaaat
     4681 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4741 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4801 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4861 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4921 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4981 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5041 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5101 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5161 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5221 ggatctcaag aagatccttt gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5281 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5341 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5401 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5461 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5521 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5581 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5641 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5701 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5761 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5821 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5881 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5941 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6001 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6061 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6121 ttattgaagc atttatcagg gttattgtct catgtccgcg cg
//
