LOCUS       Exported                9891 bp DNA     circular SYN 03-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9891)
  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..9891
                     /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..3479
                     /label=Tmpo (Lap2B) N-V5 LIC F
     misc_feature    3447..3458
                     /label=N-term LIC
     misc_feature    3459..4487
                     /label=Tmpo (Lap2B)
     primer_bind     complement(4448..4503)
                     /label=Tmpo (Lab2B) N-V5 LIC R
     misc_feature    4492..4503
                     /label=N-term LIC
     misc_feature    4531..5189
                     /label=IRES
                     /note="/vntifkey=21"
     primer_bind     complement(4710..4729)
                     /label=IRES-R Addgene
     primer_bind     complement(4782..4799)
                     /label=IRES-R Jakki
     CDS             5190..5789
                     /codon_start=1
                     /label=Puro
                     /note="/vntifkey=4"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    5813..6404
                     /label=WPRE
                     /note="/vntifkey=21"
     CDS             complement(8529..9389)
                     /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 gccggagttc ctagaggacc
     3481 cttcggtcct gaccaaagac aagttgaaga gcgagttggt cgccaacaac gtgacgctcc
     3541 cggccggcga gcagcgcaag gacgtgtacg tgcagctcta cctgcagcac ctcacggcgc
     3601 gcaaccggcc gccgctcgcc gcgggagcca acagcaaagg gccgcccgac ttctcgagcg
     3661 acgaggagcg cgagcccacg ccggtgctcg gctccggggc ctccgtgggt cgcggccgcg
     3721 gcgccgtcgg caggaaagcc acaaagaaaa ctgataagcc caggctagaa gataaagatg
     3781 atctggatgt gacagagctc tctaatgaag aacttctgga tcagcttgta agatatggag
     3841 tgaatcctgg tcccattgtg ggaacaacca ggaagctata tgagaagaag ctgttgaagc
     3901 tgagggagca gggaactgaa tcgagatcct ctactcctct tccaacagtc tcttcctctg
     3961 cagaaaacac aaggcagaat ggaagtaacg actctgacag atacagcgac aatgatgaag
     4021 actctaaaat agagctgaag cttgagaaga gggagccgct aaagggcaga gccaagacgc
     4081 cagtcacact gaagcagaga aggactgagc acaatcaggt gggagaaaaa acagaggagc
     4141 gaagagtaga cagggatatt cttaaggaaa tgttccccta cgaagcctcc actccgacgg
     4201 gaatcagtgc tagctgccgc agaccaatca aaggtgctgc aggccggccg ctcgagctca
     4261 gtgacttcag gatggaagag tcgttctcat ctaagtacgt cccgaagtat gctcccttgg
     4321 cagatgtcaa gtcagaaaag acaaagaaga gacgctccgt tcccatgtgg ataaaaatgt
     4381 tgctgtttgc ccttgtggcc gtgtttttgt ttttggtcta tcaagctatg gaaaccaacc
     4441 aaggaaatcc cttcactaat tttcttcaag atactaaaat atccaactaa tgggccgcga
     4501 cgatgtttaa acttcgaaac gcgtgaattc ctcgagggcg gccgctctag agtcgacggg
     4561 ccgcggtaac aattgttaac taacttaagc tagcaacggt ttccctctag cgggatcaat
     4621 tccgcccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     4681 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     4741 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     4801 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     4861 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     4921 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     4981 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     5041 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     5101 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     5161 ggttttcctt tgaaaaacac gataatacca tgaccgagta caagcccacg gtgcgcctcg
     5221 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     5281 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     5341 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     5401 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     5461 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     5521 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5581 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5641 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5701 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5761 ggtgcatgac ccgcaagccc ggtgcctgac ccgggcgggg cgcgtctgga acaatcaacc
     5821 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5881 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5941 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     6001 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     6061 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     6121 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     6181 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     6241 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6301 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6361 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt
     6421 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
     6481 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt tttccagtca cacctcaggt
     6541 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6601 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     6661 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag
     6721 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga
     6781 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga
     6841 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt
     6901 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg
     6961 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
     7021 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     7081 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     7141 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     7201 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     7261 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac
     7321 attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     7381 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7441 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7501 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7561 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7621 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7681 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7741 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7801 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7861 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7921 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7981 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     8041 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8101 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8161 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     8221 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8281 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     8341 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     8401 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     8461 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8521 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8581 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8641 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8701 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8761 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8821 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8881 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8941 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9001 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     9061 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     9121 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     9181 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     9241 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     9301 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     9361 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     9421 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     9481 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     9541 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     9601 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     9661 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac
     9721 cccataccct attaccactg ccaattacct agtggtttca tttactctaa acctgtgatt
     9781 cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag
     9841 tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag t
//
