LOCUS       Exported               10923 bp DNA     circular SYN 25-JUN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 10923)
  AUTHORS   Liza Brusman
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10923
                     /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..3475
                     /label=Tmem214 N-term LIC V5 F
     misc_feature    3447..3458
                     /label=N-term LIC
     misc_feature    3459..5521
                     /label=Tmem214
     primer_bind     complement(5496..5535)
                     /label=Tmem214 N-term V5 LIC R
     misc_feature    5524..5535
                     /label=N-term LIC
     misc_feature    5563..6221
                     /label=IRES
                     /note="/vntifkey=21"
     primer_bind     complement(5742..5761)
                     /label=IRES-R Addgene
     primer_bind     complement(5814..5831)
                     /label=IRES-R Jakki
     CDS             6222..6821
                     /codon_start=1
                     /label=Puro
                     /note="/vntifkey=4"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    6845..7436
                     /label=WPRE
                     /note="/vntifkey=21"
     CDS             complement(9561..10421)
                     /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 ggcggccaga tcagcgggta
     3481 gcggtggctg ggaggtggtg aagaggggcc ggcgacctgg agcaagcagc ggtgggcgag
     3541 gcggcggcgg tggcagtgac cgccgggcgc tcggggaggc aaatggagtg ttgaaatacg
     3601 acctgagctc accaatccag actacaagca ctctttatga gcgtggcttt gagaaaatca
     3661 tgaaacggca gaataaagag caagttccac cccctgctgc agagtctaag aaaccgataa
     3721 acaagaagca accaaagaag gtgacagctg ttcctagcca aaaccagaaa cagggcccat
     3781 tccgacgcct ggaggacgca ctgaaagcgc tggatgtggc agctctgcag aaggaactgg
     3841 acaagagcca gagcgtgttc actgggaacc cctccgtgtg gctgaaggac ctggccagct
     3901 atctcaacta caagctccag actccacgga tggagcccac tctgagccag tatccacatg
     3961 attatcccta cagccttgtg agccgagagc tacgtgggat catccggggg ctcctgacca
     4021 aagctgcagg gtctgtggag ctcttttttg accactgtct attcaccatg ctgcaagagc
     4081 tggataaaac accaggggag tcgctacatg ggtaccgcat ctgtattcaa gccgttctgc
     4141 aagacaagcc taagattgtc acctcaaacc tggacaagtt cctagagctt ctgcggtccc
     4201 accagagccg accagcaaag tgcctgacca tcatgtgggc cctggggcaa gcgggtttca
     4261 ccaacctcac tgagggactg aaagtgtggc tggggatcat gctgcctgtg ctgggcatca
     4321 aggctctgtc tccctttgcc atcgcatacc tggaccggct gctcctgatg catcccaacc
     4381 tcaccaaagg ctttggcatg attggcccca aggacttctt cccccttctg gactttgcct
     4441 atatgcccaa caattccctg agccccagcc tgcaggagca gctgtgccag ctcttccccc
     4501 gattgaaggt gctggcattt ggggccaagc cggaatcttc cctgcacacc tacttcccgt
     4561 cattcctgtc cagagccacc cctagctgtc ctgctgccat gaagaaagag ctcctggcca
     4621 gcctgaccca gtgcctgact gtggaccccc ttagcactag tgtctggaga cagctgtacc
     4681 ccaagcatct gtcacagtcc agcctgctgc tggagcacct actcaagtcc tgggagcaca
     4741 tccccaagaa ggcacggaaa tctttacaag aaaccatcca gtccttgaag gttaccaacc
     4801 aggagcttct gaagaagggc agtggtggca gtgagcatgt tctcacctgt gacacagcct
     4861 gcaagggctt gttgcaacgg gcacggggtc ctcggccacc ctgggcccgg ctattcctgc
     4921 tgcttctggt ctttgcagta ggattcctat gtcatgactt gcggtcaaac agctctttgc
     4981 aggcctccct cactggccgg ctgcttcgat cctccggtct cttgcctgtt ggccagcaag
     5041 tgtgtgcgag actctcctcc tacagcctgc agagctacaa ctggctgcag gagaccttgc
     5101 cagcctgtgg ctcccacctg cttgctgtgg tacagcccag tctgcagctg gcctggacac
     5161 acatctatgc catattcagc ttcctttctg cccactgtgc ctcctacctt gcctgcttca
     5221 gtgatagcct ggctggcttc tttcagaggg tccagctccc tgaggccctg cagcagctct
     5281 tccacgcctt gaaggaactg ctgctgctct tctgccatag cgtgctgctg cccacatggc
     5341 acttgctgct tgcagccctg gcccaagtcc aggagcattg ccatgaggcc tgcagagggg
     5401 acgtgacctg ggactgcatt aagacacagt tgagcagagc tgcccaatgg acctggctct
     5461 gcctgcagga tgtcacagtg gctttcttgg actgggcact caccatgata tcccagcaat
     5521 aatgggccgc gacgatgttt aaacttcgaa acgcgtgaat tcctcgaggg cggccgctct
     5581 agagtcgacg ggccgcggta acaattgtta actaacttaa gctagcaacg gtttccctct
     5641 agcgggatca attccgcccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     5701 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     5761 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     5821 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     5881 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     5941 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     6001 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     6061 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     6121 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     6181 ccacggggac gtggttttcc tttgaaaaac acgataatac catgaccgag tacaagccca
     6241 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     6301 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     6361 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     6421 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     6481 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     6541 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     6601 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     6661 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     6721 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     6781 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acccgggcgg ggcgcgtctg
     6841 gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     6901 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     6961 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     7021 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     7081 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     7141 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     7201 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     7261 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     7321 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     7381 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat
     7441 taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct
     7501 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg tttttccagt
     7561 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     7621 tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct
     7681 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
     7741 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
     7801 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
     7861 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
     7921 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
     7981 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
     8041 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
     8101 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     8161 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     8221 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     8281 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     8341 agaggccttg acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca
     8401 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     8461 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     8521 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     8581 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     8641 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     8701 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     8761 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     8821 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     8881 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     8941 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     9001 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     9061 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     9121 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     9181 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     9241 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     9301 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     9361 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     9421 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     9481 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     9541 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     9601 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     9661 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     9721 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     9781 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9841 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9901 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9961 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    10021 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    10081 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    10141 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    10201 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    10261 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    10321 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    10381 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    10441 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    10501 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
    10561 ggatcgggag atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
    10621 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    10681 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc
    10741 caaacttccc accccatacc ctattaccac tgccaattac ctagtggttt catttactct
    10801 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
    10861 ctacaaactt agtagttttt aaagaaattg tatttgttaa atatgtacta caaacttagt
    10921 agt
//
