LOCUS       Exported               10860 bp DNA     circular SYN 21-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 10860)
  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..10860
                     /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=Lmna N-V5 LIC F
     misc_feature    3447..3458
                     /label=N-term LIC
     misc_feature    3459..5456
                     /label=Lmna
     primer_bind     complement(5433..5472)
                     /label=Lmna N-V5 LIC R
     misc_feature    5461..5472
                     /label=N-term LIC
     misc_feature    5500..6158
                     /label=IRES
                     /note="/vntifkey=21"
     primer_bind     complement(5679..5698)
                     /label=IRES-R Addgene
     primer_bind     complement(5751..5768)
                     /label=IRES-R Jakki
     CDS             6159..6758
                     /codon_start=1
                     /label=Puro
                     /note="/vntifkey=4"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    6782..7373
                     /label=WPRE
                     /note="/vntifkey=21"
     CDS             complement(9498..10358)
                     /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 ggagaccccg tcacagcggc
     3481 gcgccacccg cagtggggcg caggccagct ctaccccact gtcgcccact cggatcaccc
     3541 ggctgcagga gaaggaggac ctgcaggagc tcaatgaccg cctggccgtg tacatcgatc
     3601 gcgtgcgttc cctggagacc gagaacgcgg ggctgcgcct tcgcatcact gagtctgaag
     3661 aggtggtcag ccgagaggtg tccggcatca aggcggccta cgaggccgag ctgggggatg
     3721 cccgcaagac ccttgattct gtggccaagg agcgcgcccg cctccagcta gagctgagca
     3781 aagtgcgtga ggagttcaag gagctgaagg ctcgcaacac caagaaggag ggggacttgt
     3841 tggctgcgca ggcccggctc aaggacctcg aggctcttct caactccaag gaagctgccc
     3901 tgagcactgc tctcagtgag aagcgcacat tggagggcga gctccatgac ctgcgggggc
     3961 aggtagccaa gcttgaggcg gccctgggag aggctaagaa gcagcttcag gatgagatgc
     4021 tgaggcgagt ggatgctgag aacaggctac agacgctgaa ggaggagctt gacttccaga
     4081 agaacattta cagcgaggaa ctgcgtgaga ccaagcgccg gcatgagacg cggcttgtgg
     4141 agatcgataa cgggaagcag cgagagtttg agagccggct ggcagatgcc ctgcaggagc
     4201 tgcgggctca gcatgaggac caggtggaac agtataagaa ggagctagaa aagacatact
     4261 ccgccaagct ggataatgcc aggcagtctg ctgagaggaa cagcaacctc gtgggggctg
     4321 cccatgagga actgcagcag tctcgaatcc gcattgacag cctctcggcc cagctcagcc
     4381 agctccaaaa gcagttggca gccaaggagg caaagctgcg tgacctggag gactcgctgg
     4441 cccgtgagcg cgataccagc cggcgcctgc tggctgagaa agagcgagag atggcggaga
     4501 tgcgggcgag gatgcagcag cagctggacg agtaccagga gctgctggac atcaagctgg
     4561 ccctggacat ggagatccat gcctatcgaa agctgctgga gggcgaggag gagaggctgc
     4621 gcctgtcccc cagccctacc tcgcagcgca gccgtggccg cgcctcctcc cactcatccc
     4681 agtctcaggg tggaggcagc gtcaccaaaa agcgcaagct ggagtcttcc gagagccgga
     4741 gcagcttctc gcagcatgct cgcactagcg ggcgtgtggc ggtagaggaa gtcgatgaag
     4801 agggaaagtt cgtgcggctg cgcaacaagt ccaacgagga ccagtccatg ggcaactggc
     4861 agatcaggcg tcagaatggt gacgatcctt tgatgaccta tcgcttccca ccgaagttca
     4921 ccctaaaggc tgggcaggtg gtgacgatct gggcttcagg agctggggcc acccatagcc
     4981 cccctactga cttggtgtgg aaggcgcaga acacctgggg ctgtgggagc agccttcgca
     5041 ccgctctcat caactccact ggagaagaag tggccatgcg caagctggtg cgctcactga
     5101 ccatggttga ggacaatgag gatgacgacg aggatggaga agagctcctc catcaccacc
     5161 gtggttccca ctgcagcggc tcgggggacc ccgctgagta caacctgcgc tcacgcaccg
     5221 tgctgtgcgg gacgtgtggg cagcctgctg acaaggctgc cggtggagcg ggagcccagg
     5281 tgggcggatc catctcctct ggctcttctg cctccagtgt cacagtcact cgaagcttcc
     5341 gcagtgtggg gggcagtggg ggtggcagct tcggggacaa cctagtcacc cgctcctacc
     5401 tcctgggcaa ctccagtccc cggagccaga gctcccagaa ctgcagcatc atgtaataat
     5461 gggccgcgac gatgtttaaa cttcgaaacg cgtgaattcc tcgagggcgg ccgctctaga
     5521 gtcgacgggc cgcggtaaca attgttaact aacttaagct agcaacggtt tccctctagc
     5581 gggatcaatt ccgccccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     5641 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     5701 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     5761 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     5821 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     5881 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     5941 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     6001 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     6061 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     6121 cggggacgtg gttttccttt gaaaaacacg ataataccat gaccgagtac aagcccacgg
     6181 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     6241 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     6301 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     6361 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     6421 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     6481 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     6541 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     6601 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     6661 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     6721 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc cgggcggggc gcgtctggaa
     6781 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     6841 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6901 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6961 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     7021 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     7081 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     7141 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     7201 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     7261 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     7321 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa
     7381 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     7441 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt ttccagtcac
     7501 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     7561 aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     7621 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
     7681 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     7741 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     7801 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     7861 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
     7921 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
     7981 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
     8041 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     8101 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     8161 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     8221 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     8281 ggccttgaca ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg
     8341 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     8401 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     8461 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     8521 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     8581 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     8641 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     8701 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     8761 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8821 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8881 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8941 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     9001 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     9061 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     9121 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     9181 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     9241 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
     9301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     9361 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     9421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     9481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     9541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     9601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     9661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     9721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     9781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     9841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    10021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    10081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    10141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    10201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    10261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    10321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    10381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    10441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
    10501 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
    10561 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
    10621 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
    10681 acttcccacc ccatacccta ttaccactgc caattaccta gtggtttcat ttactctaaa
    10741 cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta
    10801 caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
