LOCUS       Exported                9393 bp DNA     circular SYN 24-JUL-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9393)
  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..9393
                     /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=Vkorc1l1 N-V5 LIC F
     misc_feature    3447..3458
                     /label=N-term LIC
     primer_bind     3457..3473
                     /label=Vkorc1l1 C-V5 LIC F
     CDS             3462..3992
                     /codon_start=1
                     /label=Vkorc1l1
                     /translation="MAAPVLLRVSVPRWERVARYAVCAAGILLSIYAYHVEREKERDPE
                     HRALCDLGPWVKCSAALASRWGRGFGLLGSIFGKDGVLNQPNSVFGLIFYILQLLLGMT
                     ASAVAALVLMTSSIVSVVGSLYLAYILYFVLKEFCIICVTTYVLNFLLLIINYKRLVYL
                     NEAWKRQLQPKED"
     primer_bind     complement(3969..4005)
                     /label=Vkorc1l1 N-V5 LIC R
     primer_bind     complement(3969..3989)
                     /label=Vkorc1l1 C-V5 LIC R
     misc_feature    3994..4005
                     /label=N-term LIC
     misc_feature    4033..4691
                     /label=IRES
                     /note="/vntifkey=21"
     primer_bind     complement(4212..4231)
                     /label=IRES-R Addgene
     primer_bind     complement(4284..4301)
                     /label=IRES-R Jakki
     CDS             4692..5291
                     /codon_start=1
                     /label=Puro
                     /note="/vntifkey=4"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    5315..5906
                     /label=WPRE
                     /note="/vntifkey=21"
     CDS             complement(8031..8891)
                     /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 gatggcggcg cccgtcctgc
     3481 tgagagtgtc ggtgccgcgt tgggaacggg tggcccggta tgcagtgtgc gccgccggga
     3541 tcctgctctc catctacgcc taccacgtgg agcgggagaa ggagagggac ccggagcacc
     3601 gggccctctg cgacctgggg ccctgggtga agtgctccgc cgccctggcc tccagatggg
     3661 gtcgaggatt tggtcttttg ggttccattt ttggaaaaga tggtgtatta aaccagccaa
     3721 acagtgtctt tggacttata ttttatatac tacagttatt acttggcatg acagccagcg
     3781 cagttgcagc tctggtcctc atgacctcct ccattgtgtc tgtggtgggc tctttgtacc
     3841 tggcctacat tctgtacttt gtgctgaaag agttttgcat catctgcgtc accacatatg
     3901 tgctgaactt cctcctcctc atcatcaatt acaaacgact agtttatttg aatgaggcct
     3961 ggaagcgaca gctgcagcct aaggaagact aatgggccgc gacgatgttt aaacttcgaa
     4021 acgcgtgaat tcctcgaggg cggccgctct agagtcgacg ggccgcggta acaattgtta
     4081 actaacttaa gctagcaacg gtttccctct agcgggatca attccgcccc ccccccctaa
     4141 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     4201 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     4261 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     4321 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     4381 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     4441 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     4501 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     4561 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     4621 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     4681 acgataatac catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4741 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4801 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4861 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4921 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4981 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5041 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     5101 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     5161 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     5221 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     5281 ccggtgcctg acccgggcgg ggcgcgtctg gaacaatcaa cctctggatt acaaaatttg
     5341 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5401 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5461 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5521 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5581 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5641 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5701 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5761 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5821 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5881 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat
     5941 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     6001 caagaggagg aggaggtggg tttttccagt cacacctcag gtacctttaa gaccaatgac
     6061 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct
     6121 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     6181 cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg
     6241 atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga
     6301 gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     6361 gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc
     6421 ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt
     6481 tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata
     6541 agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6601 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6661 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6721 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6781 aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag cgttttaccg
     6841 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6901 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6961 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7021 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7081 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7141 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7201 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7261 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7441 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7681 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7801 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7861 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7921 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7981 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8041 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8101 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8161 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8221 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8281 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8341 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8401 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8461 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8521 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8581 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8641 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8701 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8761 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8821 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8881 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8941 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9001 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc
     9061 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     9121 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa
     9181 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac
     9241 tgccaattac ctagtggttt catttactct aaacctgtga ttcctctgaa ttattttcat
     9301 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttttt aaagaaattg
     9361 tatttgttaa atatgtacta caaacttagt agt
//
