LOCUS       sequence-222160-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRNA synthesis of NLS-TIR1-HA for injection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5891)
  AUTHORS   Daniel K, Icha J, Horenburg C, Muller D, Norden C, Mansfeld J
  TITLE     Conditional control of fluorescent protein degradation by an 
            auxin-dependent nanobody.
  JOURNAL   Nat Commun. 2018 Aug 17;9(1):3297. doi: 10.1038/s41467-018-05855-5.
  PUBMED    30120238
REFERENCE   2  (bases 1 to 5891)
  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"; journalName: "Nat 
            Commun."; date: "2018-08-17"; volume: "9(1):3297. doi"; pages: " 
            10.1038/s41467-018-05855-5"
COMMENT     Sequence Label: pCS2+_NLS-Tir1-HA
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      100..109
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             121..141
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             148..1869
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="TYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFVG
                     NCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEEL
                     RMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEVE
                     DRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTLA
                     KILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYPL
                     CAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELRV
                     FPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRLC
                     ILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSIA
                     FAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKGC
                     QVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     CDS             1873..1899
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1940..2074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1964..1983)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2018..2037
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2187..2207)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2187..2205)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2226..2242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2226..2242)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2239..2261)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2250..2266
                     /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(2274..2280,2281..2298,2299..2304))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2319..2340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2457..2474)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2628..3216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2708..2727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3387..4178,4179..4247))
                     /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     4010..4029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4248..4352)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4378..4833)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4515..4536)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4727..4746
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4907..5891
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttacccgccg ccaccatggg ccccgccgcc
      121 cccaagaaga agagaaaggt gctggacacg tacttccctg aagaagttgt ggaacatatc
      181 ttttcttttc ttcctgccca aagagaccgg aatacagtct ctttggtatg taaggtttgg
      241 tatgagattg aaaggttgtc aaggagagga gtgtttgtgg gaaactgcta tgcggtaagg
      301 gctggaagag tcgctgccag attccccaat gtgagggcac ttacagttaa gggtaaacca
      361 cattttgctg atttcaattt ggttcctccg gactggggag gttatgctgg accatggatt
      421 gaggctgctg ccagaggttg tcatgggttg gaggagttga ggatgaagag gatggttgtg
      481 tctgatgagt cccttgaact tctggcccgg tccttcccca gattcagagc tttggtcttg
      541 atttcttgtg agggtttctc tactgacgga ctcgctgcag tcgcttccca ctgtaagttg
      601 ttgagggagt tggacctcca agaaaatgag gttgaggaca gaggaccaag atggttgtca
      661 tgtttccctg actcatgtac ttcccttgtt tccttgaatt tcgcgtgtat caaaggagag
      721 gtcaatgccg ggagtcttga gagattggtc tcaagaagtc ccaacttgcg atcactgagg
      781 ctcaacagat ctgtgagtgt ggataccttg gcaaagattc tcttgagaac ccccaatttg
      841 gaggaccttg gaactgggaa tctcacggat gacttccaaa ctgagtctta ctttaagttg
      901 actagtgctt tggaaaagtg taagatgttg agatcactgt ccggattctg ggatgcttca
      961 cctgtttgtc tttccttcat ctaccctctt tgtgcccagc tgactggact caacttgtcc
     1021 tatgctccaa cactggatgc gagtgacttg accaagatga tttcaagatg cgtgaagctt
     1081 caaagacttt gggtcctcga ttgtatctcc gacaaaggac ttcaagtggt ggcatcatct
     1141 tgtaaggatc tccaagagtt gagagtgttt ccaagtgact tctatgtggc aggttacagt
     1201 gctgtaactg aggaagggtt ggttgctgtc tcccttggat gtcccaagtt gaattcactc
     1261 ttgtactttt gtcaccaaat gactaacgct gcccttgtga cagtggccaa aaattgccct
     1321 aacttcacac gcttcagact ttgtatcctt gaacctggga aacctgacgt ggtcaccagt
     1381 cagcccttgg acgaaggatt tggggcgatt gtgagagaat gtaagggact ccaaagactc
     1441 tctatcagtg gactcctgac tgacaaggtg tttatgtaca ttggtaagta cgctaagcag
     1501 ctggaaatgc tgtctattgc ctttgccggt gattctgaca aggggatgat gcatgttatg
     1561 aatggatgta agaacttgag aaagcttgag ataagagaca gtccattcgg agatgcagcc
     1621 ctccttggta acttcgcaag atacgaaact atgaggtctc tttggatgtc cagctgtaat
     1681 gttacactca agggatgtca agtgctggcc tcaaaaatgc caatgctgaa tgtggaggtg
     1741 atcaacgaga gagatggatc caatgaaatg gaagagaacc acggagatct tcccaaggtg
     1801 gaaaagttgt atgtctacag aaccactgct ggggcaaggg atgacgctcc aaacttcgtg
     1861 aaaatactgg gctacccata cgatgttcct gactatgcgt gactcgagcc tctagaacta
     1921 tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt ttggacaaac
     1981 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2041 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2101 gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca
     2161 ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     2221 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     2281 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     2341 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2401 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2461 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2521 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2581 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2641 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2701 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2761 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2821 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2881 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2941 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3001 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3061 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3121 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3181 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3241 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3301 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3361 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3421 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3481 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3541 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3601 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3661 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3721 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3781 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3841 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3901 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3961 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4021 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4081 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4141 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4201 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4261 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4321 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     4381 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4441 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4501 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4561 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4621 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4681 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4741 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4801 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     4861 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc atagccaatt
     4921 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
     4981 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
     5041 caatatggcg gacttggcac catgccaatt caatatggcg gacttggcac tgtgccaact
     5101 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
     5161 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
     5221 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
     5281 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
     5341 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
     5401 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
     5461 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
     5521 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
     5581 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
     5641 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
     5701 aggacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
     5761 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
     5821 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
     5881 gtttagggaa c
//
