LOCUS       Exported                7976 bp ds-DNA     circular SYN 05-OCT-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7976)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 10, 2021 from SnapGene 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7976
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=pF-PmrA-VSVG"
                     /note="color: #ffffff"
     source          601..800
                     /organism="synthetic DNA construct"
                     /label=Francisella tularensis subsp. holarctica
                     /note="color: #000000"
     promoter        299..533
                     /note="GroE"
                     /note="color: #ffffff"
     primer_bind     361..388
                     /label=KROL336 pF_seq_NEW2
                     /note="color: black; sequence: 
                     ttaaagttaaaatcgagagcttgtttga"
     primer_bind     424..450
                     /label=KROL335 pF_seq_NEW1
                     /note="color: black; sequence: ttgactcaatatctagacttgcaagag"
     misc_feature    436..458
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="AROL2"
                     /note="color: #a6acb3"
     primer_bind     585..625
                     /label=KROL39 OErpsU2F
                     /note="color: black; sequence: 
                     atatGAATTCaggaggagggtatgccaagcgttagaattaa"
     CDS             606..800
                     /codon_start=1
                     /label=30S ribosomal protein S21
                     /note="a small basic protein that is one of the last in the
                     subunit assembly; omission does not prevent assembly but 
                     the subunit is inactive; binds central domain of 16S rRNA"
                     /note="color: #ff0000"
                     /translation="MPSVRIKEREPFDVALRRFKRSCEKAGIVSELRRREYFEKPTWAR
                     KRKKTAAVKRAHKSNIIVKR"
     gene            606..800
                     /label=rpsU
                     /note="color: #00e600; direction: RIGHT"
     primer_bind     complement(772..808)
                     /label=KROL40 OE_rpsU2R
                     /note="color: black; sequence: 
                     atatGCGGCCGCccttttaactattatattgctcttatgag"
     primer_bind     801..837
                     /label=KROL333  TAP_tag_forward_notI
                     /note="color: black; sequence: 
                     atatgcGGCCGCCAAGAGAAGATGGAAAAAGAATTTCATAG"
     CDS             810..1352
                     /label=TAP
                     /label=rpoC-TAP
                     /note="color: #993366; direction: RIGHT"
     CDS             810..887
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /note="color: #cc99b2"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             915..935
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /note="color: #cc99b2
                     Cleavage site after base 932"
                     /translation="ENLYFQG"
     primer_bind     complement(944..965)
                     /label=KROL218 rpoC-TAP-ckR
                     /note="color: black; sequence: GTGTTGCGCAAGAGCCGCGGTT"
     CDS             975..1148
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /note="color: #cc99b2"
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1152..1322
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /note="color: #cc99b2"
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     primer_bind     complement(1333..1362)
                     /label=KROL334  TAP_tag_reverse_bamHI
                     /note="color: black; sequence: 
                     ATATGGATCCTCAGGTTGACTTCCCCGCGGAAT"
     misc_feature    complement(1481..1503)
                     /label=PCR_primer
                     /label=nonstandard type:  PCR_primer
                     /note="AROL3"
                     /note="color: #a6acb3; direction: LEFT"
     gene            1646..2521
                     /label=ORF3
                     /note="ORF3"
                     /note="color: #a6acb3; direction: RIGHT"
     gene            complement(2744..3163)
                     /label=ORF2
                     /note="ORF2"
                     /note="color: #a6acb3; direction: LEFT"
     gene            complement(3169..4185)
                     /label=RepA
                     /note="RepA"
                     /note="color: #a6acb3; direction: LEFT"
     promoter        5226..5275
                     /note="pCR2.1 NEOKAN promoter"
                     /note="color: #ffffff"
     gene            5364..6155
                     /label=kanamycin
                     /note="kanamycin"
                     /note="color: #a6acb3; direction: RIGHT"
     gene            6176..7033
                     /label=ampicillin
                     /note="ampicillin"
                     /note="color: #a6acb3; direction: RIGHT"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccttgta tggattagtc gagctaaaaa gctcatattt tttatattca aactatatac
      301 ccttcaagct ttgaaaaata aacttaatta ttatatatgt tatttagcta gtttttttaa
      361 ttaaagttaa aatcgagagc ttgtttgaca aaaaaacaaa aaaatttctt gaaaattttt
      421 tttttgactc aatatctaga cttgcaagag cttggaactt tgagattgtt ctaagatgca
      481 tacaaattca aaatgcttaa acaaaaataa tttaacaaag gagtaagatt gttatgaaca
      541 ttcgtccatt acaagataga gtattagttc gtcgtgcagg taccatatga attcaggagg
      601 agggtatgcc aagcgttaga attaaagaaa gagaaccttt tgatgtcgct cttagaagat
      661 ttaaaagatc ttgcgaaaag gctggaatag tatctgaact acgtcgtaga gaatactttg
      721 aaaagccaac ttgggcacgc aaaagaaaaa aaactgccgc tgttaaaaga gctcataaga
      781 gcaatataat agttaaaagg gcggccgcca agagaagatg gaaaaagaat ttcatagccg
      841 tctcagcagc caaccgcttt aagaaaatct catcctccgg ggcacttgat tatgatattc
      901 caactactgc tagcgagaat ttgtattttc agggtgagct caaaaccgcg gctcttgcgc
      961 aacacgatga agccgtggac aacaaattca acaaagaaca acaaaacgcg ttctatgaga
     1021 tcttacattt acctaactta aacgaagaac aacgaaacgc cttcatccaa agtttaaaag
     1081 atgacccaag ccaaagcgct aaccttttag cagaagctaa aaagctaaat gatgctcagg
     1141 cgccgaaagt agacaacaaa ttcaacaaag aacaacaaaa cgcgttctat gagatcttac
     1201 atttacctaa cttaaacgaa gaacaacgaa acgccttcat ccaaagttta aaagatgacc
     1261 caagccaaag cgctaacctt ttagcagaag ctaaaaagct aaatggtgct caggcgccga
     1321 aagtagacgc gaattccgcg gggaagtcaa cctgaggatc cactagctcg tttcaaatta
     1381 ccgatgatat cggaccgttc caacttaccg accagttcgg caggtatgta tttgcgtgca
     1441 ttcctatcca aaaaaacatc aagccaaaag cttgaaaaaa cttacaacac agctcaacag
     1501 agctagattg taaaaccctg ctttgttaag cagaacgcaa aaattgaatg acttatagtc
     1561 atatcgcttc gaccctcgta gattagtagc cttgagctat taactggttg aaacacttac
     1621 caaataaaga ttaaaagcga taaaaatgaa agataaagca gctaaaaaca gagattttag
     1681 aaagactatt ttatcagtgt tacaacgcaa taaagatggc tcttttgcta cgcaagcaaa
     1741 tagaaagtct attctgttgc aggcaactaa agaccttaaa aaggtagggt ttagcaaggt
     1801 tacagccgaa aacttcggta ataagcattg ctatgcactt agagaccatt ggagagccca
     1861 aggattagct acagcaacga taaaaaatcg tttagcttgt ctaaggtggt taggcgagaa
     1921 aatgggcaaa gaactacccg ataatcgaaa attagagatt gagaacagga agtatagcga
     1981 taattcaatc aataaagccc aagaaatcga ttttaaggcg atttctgcct taactgatag
     2041 gcaagcccta gcaatacaat tacagcgcga atttgggctt cgtagagaag aaagtttgaa
     2101 gtttcagcct agttatgcaa tcaaagagca taaaatcgag cttaaaagct cttggacaaa
     2161 gggtggaaga ccacgagaaa tcccaatttt gaatgaaaaa cagagagaat tgttagaaaa
     2221 agtaaaagag gtagcaggta aaggctctct aattgagagc gaaaagtctt ataagcaagc
     2281 aatggaacat ttcacgactc gctgtcaaag agcagggatt aagaatgttc atggctttag
     2341 acatgcgtat gctcaagata gatataggca attaacaggg cgtgagtgtc ctaaaaatgg
     2401 tggattaaca tctaagcagc taacacctga gcaaaagcaa caagactatg aagctagaat
     2461 gactattagt gaagagttag gtcatggtag agaagatgta acagtcaact acttaggcag
     2521 ataaaaagca atatagctat agaagaaaag aaagctattt tacatagtag atcgactctt
     2581 cttagggatt ttatattttt tgataaatca tctattttgc tagttaaatc atcaaattta
     2641 tcatcttgtt gtttgactaa atctaagaat ctattctctt ttttaaaatc gttcatgcaa
     2701 accgcctata gctttcttct ttttctgaaa ttatttgtct tcacaccata attaaattcc
     2761 catttttata agtaaagtct tttaaaagct tgtcagtctc ttctctagaa atgtaccaaa
     2821 ttttacctat tttaggatac ttttcatgaa gctcttctat ttttccccag tcctttaata
     2881 gtctaccttt agagtctcgt aaatagttat ctttgtgaca ggggcctctt ttatcttttt
     2941 taatgtaact atatgttatt ccaacgtcac tattactatt atccaaatct tttttagcat
     3001 gccagtaaga actttcataa cttaactcta tctttcgacc tctttgatat acaacaataa
     3061 agctatagcc agtagtaaca acctgtttta cttttgttaa atctattaac ttcttattta
     3121 tttttttatg tttttttgaa aatttaaata tttctatatt cattcctaca cttcctcaaa
     3181 tccaaatggt agcttatgat tctcttctgg tttcttttct aattttttta tatttgcaat
     3241 aaaaactctt tttctatctt tgattttttt attgtcccaa ttcctccaag tatcatcaca
     3301 aaccctttca atatcatgta aatgatgatg tctaaatatt gatctgacat aatacagatc
     3361 taggtctagt tcatcactta acacaacttc tctaagtctt tcagatgctt cgattggtat
     3421 gtaatcctct ttatttttag tatctaaaag cttttgctta aattcttctt ctgtctctgc
     3481 taccttacta actgtaaact tgatatttgt aatcttacga ccatgttttc tgtgatgatc
     3541 cttgtcatca taggttacaa aaatatccga taattgatta atctcttcta gtgctggtaa
     3601 taggaactta tttttaaaat ttgaatatct gttgctgtaa cttttaggta aatcaaaatc
     3661 attaatcata tcatcgacat acaatacgca atcaactata ttagcatacc ctgcttgttc
     3721 gcctaatttg cttttgagaa gtaagtataa tctgcttgaa tacttacttt taaatgaaaa
     3781 tagtaactgt ctttctgctt tagtaaagta ctcttgtagt tgtatcatgt gtggcattaa
     3841 tgaccaatga aactcgcaaa ttaaagcact gcttttaggg tctgcttcaa tatatgcaaa
     3901 ccagttagct atcttcgttt gttctttatt cagccatact ggcttagaca ttattgagtg
     3961 cattaattgc ttcaatctca ctctgttatg cttaacccct gtagcttttt caagatcaga
     4021 taggcttatc ttatacctgt gaaactcttt atcttctctt ttaaccattg aggcaactaa
     4081 gaatattaag ttttgttctt cttttgtaag gctatacttt cctgcaacaa gagtattaga
     4141 catagctatt tctttgccag catttacatt tttaacttct ttcatagaac tagagtcatt
     4201 atctcgatat acaaattcta taaaacttct attagtaaaa caactacttc ataaaaaaaa
     4261 gtagttttaa cgatacaaaa agtagtttta aattcaaaaa gtgatacaaa aagtagtttt
     4321 aaattcaaaa agtgatacaa aaagtagttt taaatttttt aaaaaagtgc ttcaaagcct
     4381 tatgtagcaa tacttacaga ggattaaaaa aaaatctgac aatatataaa gagaatatat
     4441 aaagagaata tcttagggga ttttaaaaaa atcccacaga ctcaaagact tttttgactt
     4501 tttaaatcct agaaactata ctttaagtac ttatttaagt acatggattt agattatgca
     4561 aaccgttaat tattcaactt ttagaaatga actatctgat tcaatggata gagtaacaaa
     4621 aaatcatagt cctatgattg taactagagg ttcaaaaaaa gaagcagttg ttatgatgtc
     4681 gttagaggat tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4741 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4801 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4861 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4921 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4981 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat tcagggcgca
     5041 agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc
     5101 ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc
     5161 aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa
     5221 gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa
     5281 actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag
     5341 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5401 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5461 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5521 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5581 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5641 tattgggcga agtgccgggg caggatctcc tgtcatccca ccttgctcct gccgagaaag
     5701 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5761 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5821 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5881 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5941 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6001 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6061 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6121 gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat tgaaaaagga agagtatgag
     6181 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6241 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6301 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6361 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6421 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6481 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6541 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6601 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6661 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6721 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6781 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6841 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6901 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6961 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7021 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7081 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7141 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7201 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7261 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7321 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7381 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7441 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7501 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7561 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7621 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7681 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7741 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7801 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7861 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7921 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
