LOCUS       Exported               10634 bp DNA     circular SYN 12-FEB-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 10634)
  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..10634
                     /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..3473
                     /label=Lmnb1 N-V5 LIC F
     misc_feature    3447..3458
                     /label=N-term LIC
     misc_feature    3459..5225
                     /label=Lmnb1
     primer_bind     4140..4159
                     /label=Lmnb1 Sequencing Primer F
     primer_bind     complement(5208..5246)
                     /label=Lmnb1 N-V5 LIC R
     misc_feature    5235..5246
                     /label=N-term LIC
     misc_feature    5274..5932
                     /label=IRES
                     /note="/vntifkey=21"
     primer_bind     complement(5453..5472)
                     /label=IRES-R Addgene
     primer_bind     complement(5525..5542)
                     /label=IRES-R Jakki
     CDS             5933..6532
                     /codon_start=1
                     /label=Puro
                     /note="/vntifkey=4"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    6556..7147
                     /label=WPRE
                     /note="/vntifkey=21"
     CDS             complement(9272..10132)
                     /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 ggcgaccgcg acccccgtgc
     3481 agcagcagcg ggcgggcagc cgcgccagcg cccccgccac gccgctcagc cccacgcgcc
     3541 tgtcgcgcct gcaggagaaa gaggagctgc gggagctcaa cgaccgcctg gctgtgtaca
     3601 tcgataaggt gcgcagcctg gagacggaga acagcgcgct gcagctgcag gtgaccgagc
     3661 gcgaggaggt gcgcggccgc gagctcaccg gcctcaaggc tctctacgag accgagctgg
     3721 ccgacgcacg ccgcgctctg gacgacacgg cccgcgagcg cgccaagctc cagatcgagc
     3781 tgggcaagtt caaggccgag cacgaccagc tgctgctcaa ttatgccaag aaggaatctg
     3841 atctcagtgg agcccagatc aagcttcgag agtatgaggc ggcactaaac tctaaggatg
     3901 cggcgctggc tactgcccta ggggacaaaa agagtttaga gggagacttg gaggatctga
     3961 aagatcagat tgcccagcta gaagcatcct tatctgccgc caaaaagcag ttagcagatg
     4021 aaactttact taaagtggat ttggagaatc gctgtcagag ccttactgag gacttggagt
     4081 ttcgtaaaaa tatgtatgaa gaggagatca atgagacaag gaggaagcat gagacccgct
     4141 tggtggaagt ggactctggg cgtcagattg agtatgagta caagctggct caagccctgc
     4201 atgagatgcg ggagcagcac gacgcgcagg tgaggctgta caaggaagag ctggagcaga
     4261 cctaccacgc caagcttgag aatgccagac tctcctcaga gatgaacact tccactgtca
     4321 acagtgcccg ggaagagctg atggagagcc ggatgaggat cgagagcctc tcctcacagc
     4381 tctctaacct gcagaaagag tctagagcgt gtttggaaag gatccaggaa ttggaggaca
     4441 tgcttgctaa ggagagagac aactcgcgcc gcatgctgtc tgacagagag agagagatgg
     4501 cggagatcag ggaccagatg cagcagcagc tgagtgatta tgagcagctg ctggacgtga
     4561 agctggccct ggacatggag atcagcgcct acaggaagct cctggaaggc gaagaagagc
     4621 ggttaaagct ctctccaagc ccttcttccc gggtgaccgt gtccagagcg tcctccagtc
     4681 gcagtgtgcg caccaccaga ggaaagcgga agagagttga tgtggaggag tcggaggcga
     4741 gcagcagtgt tagcatttcc cactctgcct cagccacggg gaacgtgtgc attgaagaga
     4801 tagatgttga tgggaagttt attcgcttga agaacacttc tgagcaggat caaccaatgg
     4861 gaggctggga gatgatcaga aaaattggag acacatcagt cagttacaaa tatacctcaa
     4921 gatatgtgct gaaggctggc cagactgtca cagtgtgggc tgcaaatgct ggcgtcacag
     4981 ccagccctcc aactgacctc atctggaaga accagaactc ttggggtact ggtgaagatg
     5041 tgaaggttat attgaagaat tctcagggag aggaggttgc tcagagaagc acagtcttca
     5101 agaccaccat acccgaggag gaggaggagg aggaggagga gcccatcgga gtggctgtgg
     5161 aggaggagcg tttccaccag cagggagccc caagagcatc caataaaagc tgtgccatta
     5221 tgtgaacttg taatgggccg cgacgatgtt taaacttcga aacgcgtgaa ttcctcgagg
     5281 gcggccgctc tagagtcgac gggccgcggt aacaattgtt aactaactta agctagcaac
     5341 ggtttccctc tagcgggatc aattccgccc ccccccccta acgttactgg ccgaagccgc
     5401 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     5461 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     5521 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     5581 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     5641 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     5701 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     5761 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     5821 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     5881 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgataata ccatgaccga
     5941 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     6001 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     6061 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     6121 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     6181 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     6241 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     6301 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     6361 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     6421 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     6481 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacccgggcg
     6541 gggcgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6601 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6661 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     6721 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     6781 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     6841 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     6901 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     6961 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     7021 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     7081 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     7141 ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca
     7201 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     7261 gtttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     7321 cttagccact ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga
     7381 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
     7441 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
     7501 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
     7561 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
     7621 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
     7681 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
     7741 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
     7801 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7861 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7921 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7981 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     8041 atcagagagt gagaggcctt gacattgcta gcgttttacc gtcgacctct agctagagct
     8101 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     8161 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     8221 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     8281 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     8341 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     8401 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     8461 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     8521 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     8581 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     8641 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     8701 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     8761 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     8821 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     8881 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     8941 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     9001 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     9061 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     9121 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     9181 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     9241 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9301 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     9361 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     9421 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     9481 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     9541 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     9601 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     9661 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     9721 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     9781 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9841 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9901 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9961 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    10021 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    10081 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    10141 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    10201 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    10261 ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa
    10321 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
    10381 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
    10441 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctagtggtt
    10501 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
    10561 ttaaatatgt actacaaact tagtagtttt taaagaaatt gtatttgtta aatatgtact
    10621 acaaacttag tagt
//
