LOCUS       Exported                9441 bp DNA     circular SYN 16-OCT-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9441)
  AUTHORS   Gerace Lab
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9441
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_structure  2028..2151
                     /label=cPPT
                     /note="/vntifkey=88"
     promoter        2209..3380
                     /label=EF-1a
                     /note="/vntifkey=29"
     primer_bind     3328..3348
                     /label=EF1a-F
     regulatory      3387..3395
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3396..3437
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3438..3455
                     /label=6xGLY
                     /note="6xGLY"
     primer_bind     3447..3481
                     /label=Rnf185 N-V5 LIC F
     misc_feature    3447..3458
                     /label=N-term LIC
     misc_feature    3459..4037
                     /label=Rnf185
     primer_bind     complement(4018..4053)
                     /label=Rnf185 N-V5 LIC R
     misc_feature    4042..4053
                     /label=N-term LIC
     misc_feature    4081..4739
                     /label=IRES
                     /note="/vntifkey=21"
     primer_bind     complement(4260..4279)
                     /label=IRES-R Addgene
     primer_bind     complement(4332..4349)
                     /label=IRES-R Jakki
     CDS             4740..5339
                     /codon_start=1
                     /label=Puro
                     /note="/vntifkey=4"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    5363..5954
                     /label=WPRE
                     /note="/vntifkey=21"
     CDS             complement(8079..8939)
                     /codon_start=1
                     /label=AmpR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgaggcc ctttcgtctt cactcgaggt gcccgtcagt
     2221 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2281 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2341 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2401 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2461 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     2521 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
     2581 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
     2641 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     2701 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     2761 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     2821 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     2881 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     2941 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     3001 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     3061 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     3121 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     3181 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     3241 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     3301 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     3361 ttccatttca ggtgtcgtga ggatccgccg ccatgggtaa gcctatccct aaccctctcc
     3421 tcggtctcga ttctacgggg ggcggagggg gtgggcccat ggcaagtaaa gggccttcgg
     3481 cctctgcatc cactgagaat tccaatgcag gggggcccag tggcagcagc aatggcactg
     3541 gtgagagtgg agggcaggac agcacctttg agtgcaacat atgcctggac acagccaagg
     3601 atgctgtcat cagcctgtgt ggccacctct tctgttggcc gtgtttacat cagtggttgg
     3661 agactaggcc taacagacaa gtgtgtccag tctgcaaagc tggcatcagc cgagacaaag
     3721 tcatccccct ctatggcaga ggcagcactg gacagcagga tcccagagag aagactcctc
     3781 ctcgtcctca aggccagagg ccagagccag aaaatagagg gggatttcaa ggatttggat
     3841 ttggagatgg tggctttcag atgtcttttg gaattggagc atttccattt gggatatttg
     3901 ccacagcatt taacataaat gatggacggc ctcctccagc tgtccctggg acaccccagt
     3961 atgtggatga gcagttcctg tcacgcctct tcctgtttgt cgccctggtg atcatgttct
     4021 ggctcctgat cgcctaataa tgggccgcga cgatgtttaa acttcgaaac gcgtgaattc
     4081 ctcgagggcg gccgctctag agtcgacggg ccgcggtaac aattgttaac taacttaagc
     4141 tagcaacggt ttccctctag cgggatcaat tccgcccccc ccccctaacg ttactggccg
     4201 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     4261 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     4321 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     4381 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     4441 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     4501 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     4561 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     4621 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     4681 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gataatacca
     4741 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     4801 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     4861 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     4921 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     4981 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5041 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     5101 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     5161 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     5221 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5281 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     5341 ccgggcgggg cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac
     5401 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5461 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5521 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5581 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5641 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5701 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     5761 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5821 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5881 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5941 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca
     6001 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     6061 gaggtgggtt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     6121 tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa ttcactccca
     6181 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgatta
     6241 gcagaactac acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa
     6301 gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt
     6361 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt tagagtggag
     6421 gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa
     6481 gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc cagggaggcg
     6541 tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag cagctgcttt
     6601 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6661 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6721 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6781 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6841 aatgaatatc agagagtgag aggccttgac attgctagcg ttttaccgtc gacctctagc
     6901 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     6961 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7021 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7081 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7141 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7201 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7261 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7321 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7381 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7441 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7501 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7561 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7621 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7681 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7741 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7801 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7861 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7921 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7981 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8041 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8101 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8161 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8221 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8281 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8341 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8401 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8461 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8521 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8581 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8641 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8701 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8761 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8821 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8881 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8941 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     9001 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     9061 gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta
     9121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     9181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca
     9241 agctaaccaa aatcatccca aacttcccac cccataccct attaccactg ccaattacct
     9301 agtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt
     9361 gtatttgtta aatatgtact acaaacttag tagtttttaa agaaattgta tttgttaaat
     9421 atgtactaca aacttagtag t
//
