LOCUS       Exported                9353 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral expression of human TDP-43 D169G disease mutant in 
            mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    pLVX-Puro-TDP-43-D169G
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9353)
  AUTHORS   Roczniak-Ferguson A, Ferguson SM
  TITLE     Pleiotropic requirements for human TDP-43 in the regulation of cell 
            and organelle homeostasis.
  JOURNAL   Life Sci Alliance. 2019 Sep 16;2(5). pii: 2/5/e201900358. doi: 
            10.26508/lsa.201900358. Print 2019 Oct.
  PUBMED    31527135
REFERENCE   2  (bases 1 to 9353)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9353
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        2201..2504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2505..2708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2659..2679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2705..2729
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4141..4640
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4176..4199)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4573..4592
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4661..5260
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4661..4680)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5157..5177
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5274..5862
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5327..5347)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5745..5756)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6069..6702
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6831..6847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6831..6847)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6844..6866)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6855..6871
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6879..6909)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6924..6945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7062..7079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7233..7821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7313..7332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7992..8852)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8615..8634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8853..8957)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    9005..9139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9042..9061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9096..9115
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
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 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gccaccatgt ctgaatatat
     2881 tcgggtaacc gaagatgaga acgatgagcc cattgaaata ccatcggaag acgatgggac
     2941 ggtgctgctc tccacggtta cagcccagtt tccaggggcg tgtgggcttc gctacaggaa
     3001 tccagtgtct cagtgtatga gaggtgtccg gctggtagaa ggaattctgc atgccccaga
     3061 tgctggctgg ggaaatctgg tgtatgttgt caactatcca aaagataaca aaagaaaaat
     3121 ggatgagaca gatgcttcat cagcagtgaa agtgaaaaga gcagtccaga aaacatccga
     3181 tttaatagtg ttgggtctcc catggaaaac aaccgaacag gacctgaaag agtattttag
     3241 tacctttgga gaagttctta tggtgcaggt caagaaagat cttaagactg gtcattcaaa
     3301 ggggtttggc tttgttcgtt ttacggaata tgaaacacaa gtgaaagtaa tgtcacagcg
     3361 acatatgata ggtggacgat ggtgtgactg caaacttcct aattctaagc aaagccaaga
     3421 tgagcctttg agaagcagaa aagtgtttgt ggggcgctgt acagaggaca tgactgagga
     3481 tgagctgcgg gagttcttct ctcagtacgg ggatgtgatg gatgtcttca tccccaagcc
     3541 attcagggcc tttgcctttg ttacatttgc agatgatcag attgcgcagt ctctttgtgg
     3601 agaggacttg atcattaaag gaatcagcgt tcatatatcc aatgccgaac ctaagcacaa
     3661 tagcaataga cagttagaaa gaagtggaag atttggtggt aatccaggtg gctttgggaa
     3721 tcagggtgga tttggtaata gcagaggggg tggagctggt ttgggaaaca atcaaggtag
     3781 taatatgggt ggtgggatga actttggtgc gttcagcatt aatccagcca tgatggctgc
     3841 cgcccaggca gcactacaga gcagttgggg tatgatgggc atgttagcca gccagcagaa
     3901 ccagtcaggc ccatcgggta ataaccaaaa ccaaggcaac atgcagaggg agccaaacca
     3961 ggccttcggt tctggaaata actcttatag tggctctaat tctggtgcag caattggttg
     4021 gggatcagca tccaatgcag ggtcgggcag tggttttaat ggaggctttg gctcaagcat
     4081 ggattctaag tcttctggct ggggaatgta ggggatcccg cgactctaga taattctacc
     4141 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     4201 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     4261 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     4321 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     4381 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     4441 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     4501 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     4561 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     4621 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac
     4681 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4741 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4801 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4861 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4921 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4981 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5041 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5101 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5161 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5221 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac
     5281 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5341 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5401 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5461 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5521 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5581 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5641 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     5701 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5761 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     5821 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag
     5881 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     5941 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     6001 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6061 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     6121 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag
     6181 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga
     6241 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga
     6301 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt
     6361 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg
     6421 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
     6481 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     6541 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6601 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6661 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6721 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac
     6781 attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     6841 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6901 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6961 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7021 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7081 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7141 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7981 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8041 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8101 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8161 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8221 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8281 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8341 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8401 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8461 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8521 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8581 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8641 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8701 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8761 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8821 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8881 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8941 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     9001 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     9061 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     9121 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc
     9181 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga
     9241 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
     9301 agtagttttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
