LOCUS       sequence-6761-ex        2899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2899)
  AUTHORS   Kapust RB, Waugh DS
  TITLE     Controlled intracellular processing of fusion proteins by TEV 
            protease.
  JOURNAL   Protein Expr Purif. 2000 Jul . 19(2):312-8.
  PUBMED    10873547
REFERENCE   2  (bases 1 to 2899)
  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: "Protein 
            Expr Purif. 2000 Jul . 19(2):312-8."
COMMENT     Sequence Label: pRK603
FEATURES             Location/Qualifiers
     source          1..2899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..61
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(82..100,101..106,107..123,124..129,130..155)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    82..100
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    107..125
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             165..176
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             186..893
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMDKPE
                     EPFQPVKEATQLMN"
     terminator      929..1015
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1051..1068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1185..1730)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1265..1284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1844..1938)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1969..2763)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2080..2099)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2690..2709
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
       61 ccctttcgtc ttcacctcga gtccctatca gtgatagaga ttgacatccc tatcagtgat
      121 agagatactg agcacatcag caggacgcac tgaccgaatt cattaaagag gagaaaggta
      181 ccatgggaga aagcttgttt aaggggccgc gtgattacaa cccgatatcg agcaccattt
      241 gtcatttgac gaatgaatct gatgggcaca caacatcgtt gtatggtatt ggatttgggc
      301 ccttcatcat tacaaacaag cacttgttta gaagaaataa tggaacactg ttggtccaat
      361 cactacatgg tgtattcaag gtcaagaaca ccacgacttt gcaacaacac ctcatcgatg
      421 ggcgggacat gatcattatt cgaatgccta aggatttccc accatttcct caaaagctga
      481 aatttcgcga gccacaacgg gaagagcgca tttgtcttgt gacaaccaac ttccaaacta
      541 agagcatgtc tagcatggtg tcagacacta gttgcacatt cccttcatct gatggcatat
      601 tctggaagca ttggattcaa accaaggatg ggcagtgtgg cagtccatta gtatcaactc
      661 gagatgggtt cattgttggt atacactcag catcgaattt caccaacaca aacaattatt
      721 tcacaagcgt gccgaaaaac ttcatggaat tgttgacaaa tcaggaggcg cagcagtggg
      781 ttagtggttg gcgattaaat gctgactcag tattgtgggg gggccataaa gttttcatgg
      841 acaaacctga agagcctttt cagccagtta aggaagcgac tcaactcatg aattaataag
      901 gatcccatgg tacgcgtgct agaggcatca aataaaacga aaggctcagt cgaaagactg
      961 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     1021 gccctagacc taggggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
     1081 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     1141 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     1201 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
     1261 ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc
     1321 tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac
     1381 actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca
     1441 gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc
     1501 aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat
     1561 gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt
     1621 tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg
     1681 ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc
     1741 ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac
     1801 ctgaagtcag ccccatacga tataagttgt tactagtgct tggattctca ccaataaaaa
     1861 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     1921 gatctatcaa caggagtcca agcgagctct cgaaccccag agtcccgctc agaagaactc
     1981 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     2041 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     2101 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     2161 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     2221 gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     2281 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     2341 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     2401 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     2461 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     2521 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     2581 ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     2641 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     2701 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     2761 catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc gccatcagat
     2821 ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct taccagaggg
     2881 cgccccagct ggcaattcc
//
