LOCUS       sequence-318058-        3790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pUPD_NIaPro(TEV).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3790)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 3790)
  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"
COMMENT     Sequence Label: Tobacco Etch Virus NIa protease (GB0420)
FEATURES             Location/Qualifiers
     source          1..3790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             9..713
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMSKPE
                     EPFQPVKEATQLM"
     promoter        complement(840..858)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(841..858)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(876..892)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(876..892)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(889..911)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    900..916
                     /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(924..930,931..948,949..954))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    969..990
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1107..1124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1278..1866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1358..1377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2037..2828,2829..2897))
                     /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     2660..2679
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2898..3002)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3080..3535)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3217..3238)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3429..3448
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3661..3683
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3675..3692
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3676..3692
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3699..3718
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3699..3717
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3748..3757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3752..3761
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcgaatggg agaaagcttg tttaagggac cacgtgatta caacccgata tcgagcacca
       61 tttgtcactt gacgaatgaa tctgatgggc acacaacatc gttgtatggt attggatttg
      121 gtcccttcat cattacaaac aagcacttgt ttagaagaaa taatggaaca ctgttggtcc
      181 aatcactaca tggtgtattc aaggtcaaga acaccacgac tttgcaacaa cacctcattg
      241 atgggaggga catgataatt attcgcatgc ctaaggattt cccaccattt cctcaaaagc
      301 tgaaatttag agagccacaa agggaagagc gcatatgtct tgtgacaacc aacttccaaa
      361 ctaagagcat gtctagcatg gtgtcagaca ctagttgcac attcccttca tctgatggca
      421 tattctggaa gcattggatt caaaccaagg atgggcagtg tggcagtcca ttagtatcaa
      481 ctagagatgg gttcattgtt ggtatacact cagcatcgaa tttcaccaac acaaacaatt
      541 atttcacaag cgtgccgaaa aacttcatgg aattgttgac aaatcaggag gcgcagcagt
      601 gggttagtgg ttggcgatta aatgctgact cagtattgtg ggggggccat aaagttttca
      661 tgagcaaacc tgaagagcct tttcagccag ttaaggaagc gactcaactc atgagtgaat
      721 tggtgtactc gcaatgagct tcgagaccac tcatcgccca atcactagtg aattcgcggc
      781 cgcctgcagg tcgaccatat gggagagctc ccaacgcgtt ggatgcatag cttgagtatt
      841 ctatagtgtc acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      901 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
      961 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1021 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1081 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1141 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1201 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1261 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1321 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1381 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1441 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1501 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1561 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1621 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1681 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     1741 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1801 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1861 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1921 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1981 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2041 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2101 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2161 gctgcaatga taccgcgcga cccacgctca ccggctccag atttatcagc aataaaccag
     2221 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2281 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2341 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2401 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2461 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2521 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2581 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2641 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2701 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2761 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2821 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2881 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2941 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3001 cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag
     3061 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3121 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3181 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3241 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgctggccca
     3301 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3361 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3421 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3481 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     3541 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     3601 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     3661 cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg
     3721 aattgggccc gacgtcgcat gctcccggcc gccatggcgg ccgcgggaat tcgattgggc
     3781 gatgagtggt
//
