LOCUS       Exported               10080 bp DNA     circular SYN 26-SEP-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 10080)
  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..10080
                     /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..3469
                     /label=Chpt1 N-V5 LIC F
     misc_feature    3447..3458
                     /label=N-term LIC
     primer_bind     3457..3469
                     /label=Chpt1 C-V5 LIC F
     regulatory      3457..3466
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3459..4677
                     /codon_start=1
                     /label=Chpt1
                     /translation="MAAGAGARPAPRWVKALGEPLSAAQLRRLEEHRYTAVGESLFEPP
                     LQLYWTWLLQWIPLWMAPNTITLIGLAINLVTTLVLIFYCPTVTEEAPYWTYLLCALGL
                     FIYQSLDAIDGKQARRTNSCSPLGELFDHGCDSLSTVFMAIGASIAVRLGTHPDWLFFC
                     SFVGMFMFYCAHWQTYVSGVLRFGRVDVTEIQVALVIVFMLSTFGGATMWDYTIPILEI
                     KLKIVPVLGVVGGLIFSCSNYFHVILHGGVGKNGSTIAGTSVLSPGLHIGLIIILAIMI
                     YKKSATNMFEKHPCLYTLMFGCVFAKVAQKLVIAHMTKSELYLQDTVFIGPGLLFLDQY
                     FNNFIDEYVVLWIAMVISSFDMMIYFTSLCLQISRHLHLNIFKTSCQQAPEQVQVLSSK
                     THQNNMD"
     primer_bind     complement(4641..4692)
                     /label=Chpt1 N-V5 LIC R
     primer_bind     complement(4641..4676)
                     /label=Chpt1 C-V5 LIC R
     misc_feature    4681..4692
                     /label=N-term LIC
     misc_feature    4720..5378
                     /label=IRES
                     /note="/vntifkey=21"
     primer_bind     complement(4899..4918)
                     /label=IRES-R Addgene
     primer_bind     complement(4971..4988)
                     /label=IRES-R Jakki
     CDS             5379..5978
                     /codon_start=1
                     /label=Puro
                     /note="/vntifkey=4"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    6002..6593
                     /label=WPRE
                     /note="/vntifkey=21"
     CDS             complement(8718..9578)
                     /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 ggcggcgggc gccggggcca
     3481 ggccggcgcc gcgctgggta aaggcgctag gtgagccgct cagtgccgcg cagctgcggc
     3541 ggctggagga gcaccgctac accgcggtgg gagagtcgct gttcgagccg ccgctgcagc
     3601 tttactggac ctggctgctc caatggatcc cgctctggat ggcccccaac accatcaccc
     3661 tcatcggcct cgccatcaac ctggtcacca cactagtgct catcttctac tgccctacag
     3721 tcacggagga ggcaccatac tggacatacc ttttatgtgc cctgggactc tttatctacc
     3781 agtcactgga tgccattgat gggaaacaag ccagaaggac aaactcttgc tctcccttag
     3841 gggaactatt tgatcatggt tgtgactctc tttccacagt atttatggcc atcggcgctt
     3901 ccattgctgt tcgcctagga acacatcctg actggttgtt tttctgctct ttcgttggga
     3961 tgttcatgtt ttactgtgct cattggcaga cttacgtctc aggagtgtta agatttggaa
     4021 gggtggatgt cactgagatc caggtagctt tagtgatcgt cttcatgttg tcaacatttg
     4081 gaggagcaac aatgtgggac tatacgatac ctattctaga aataaaactg aagattgttc
     4141 cggttcttgg agttgtaggt ggattaatat tttcctgttc aaattatttc catgtgatcc
     4201 tccatggtgg tgttggcaag aatgggtcta ctattgcagg caccagtgtc ttgtcacctg
     4261 gactccacat aggattaatt attatattgg caataatgat ctataagaag tctgcaacaa
     4321 atatgtttga aaaacatcct tgcctttata ctttaatgtt tggatgtgtc tttgctaaag
     4381 ttgcacaaaa attggtgata gctcacatga cgaaaagtga actatatctt caagacactg
     4441 tctttattgg gccaggtctt ttatttttag accaatactt taataatttt atagatgaat
     4501 atgttgttct gtggatagca atggtcattt cttcatttga tatgatgata tactttactt
     4561 ctttgtgcct gcaaatttca agacaccttc atctaaacat cttcaagact tcatgtcaac
     4621 aagcaccgga acaggttcaa gttctttctt caaagactca tcagaataac atggattaat
     4681 gggccgcgac gatgtttaaa cttcgaaacg cgtgaattcc tcgagggcgg ccgctctaga
     4741 gtcgacgggc cgcggtaaca attgttaact aacttaagct agcaacggtt tccctctagc
     4801 gggatcaatt ccgccccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     4861 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     4921 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     4981 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     5041 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     5101 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     5161 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     5221 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     5281 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     5341 cggggacgtg gttttccttt gaaaaacacg ataataccat gaccgagtac aagcccacgg
     5401 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     5461 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     5521 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     5581 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     5641 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     5701 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     5761 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     5821 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     5881 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     5941 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc cgggcggggc gcgtctggaa
     6001 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     6061 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6121 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6181 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6241 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6301 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6361 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     6421 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6481 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6541 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa
     6601 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     6661 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt ttccagtcac
     6721 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     6781 aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     6841 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
     6901 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     6961 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     7021 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     7081 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
     7141 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
     7201 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
     7261 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     7321 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     7381 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     7441 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     7501 ggccttgaca ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg
     7561 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     7621 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     7681 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     7741 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7801 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
     8521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8581 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8641 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8701 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8761 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8821 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8881 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8941 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     9001 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     9061 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9121 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9181 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9241 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9301 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9361 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9421 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9481 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9541 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9601 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9661 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     9721 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     9781 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     9841 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
     9901 acttcccacc ccatacccta ttaccactgc caattaccta gtggtttcat ttactctaaa
     9961 cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta
    10021 caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
