LOCUS       Exported                7077 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  express human caspase 8 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    Fv-hCaspase 8 (C/A)-2A-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7077)
  AUTHORS   Oberst A, Pop C, Tremblay AG, Blais V, Denault JB, Salvesen GS, 
            Green DR
  TITLE     Inducible dimerization and inducible cleavage reveal a requirement 
            for both processes in caspase-8 activation.
  JOURNAL   J Biol Chem. 2010 May 28;285(22):16632-42. doi: 
            10.1074/jbc.M109.095083. Epub 2010 Mar 22.
  PUBMED    20308068
REFERENCE   2  (bases 1 to 7077)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7077
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(661..1260)
                     /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(744..764)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1241..1260
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1270..1599)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1284..1419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1338..1357)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1584..1604
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(1624..2343)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(1627..2343)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1669..1690)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2018..2037
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2277..2298
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2353..2406)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(3271..3591)
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(3598..3639)
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     CDS             complement(3653..4069)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3669..3685)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3701..3723)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4270..4469
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             4534..5003
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        5034..5138
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5139..5999
                     /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     complement(5357..5376)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6159..6747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6648..6667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6901..6918
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             7048..440
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 actcagagga gaccctccca aggaacagcg agaccacaag tcggatgcaa ctgcaagagg
       61 gtttattgga tacacgggta cccgggcgac tcagtcaatc ggaggactgg cgccccgagt
      121 gaggggttgt gggctctttt attgagctcg gggagcagaa gcgcgcgaac agaagcgaga
      181 agcgaactga ttggttagtt caaataaggc acagggtcat ttcaggtcct tggggcaccc
      241 tggaaacatc tgatggttct ctagaaactg ctgagggctg gaccgcatct ggggaccatc
      301 tgttcttggc cctgagccgg ggcaggaact gcttaccaca gatatcctgt ttggcccata
      361 ttcagctgtt ccatctgttc ttggccctga gccggggcag gaactgctta ccacagatat
      421 cctgtttggc ccatattcag ctgttccatc tgttcctgac cttgatctga acttctctat
      481 tctcagttat gtatttttcc atgccttgca aaatggcgtt acttaagcta gcttgccaaa
      541 cctacaggtg gggtctttca ttcccccctt tttctggaga ctaaataaaa tcttttattt
      601 tatcgatgca tggggtcgtg cgctcctttc ggtcgggcgc tgcgggtcgt ggggcgggcg
      661 tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc
      721 ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc
      781 caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct
      841 gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg
      901 ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga
      961 accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct
     1021 ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat
     1081 gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg
     1141 gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg
     1201 tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat
     1261 ggtaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     1321 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     1381 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     1441 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     1501 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     1561 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacaggg tcgacgttta
     1621 aacttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     1681 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag
     1741 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     1801 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     1861 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     1921 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     1981 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     2041 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     2101 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     2161 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     2221 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     2281 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     2341 cataggcgcg ccagggccgg gattctcctc cacgtcaccg catgttagaa gacttcctct
     2401 gccctctgcg atcgcatcag aagggaagac aagttttttt cttagtgtga aagtaggctg
     2461 aggcatctgt ttccccatgt ttttcttgtc atccttgttg cttacttcat agttcacttc
     2521 agtcaggatg gtgagaatat catcgcctcg agggcatcgc tctctcaggc tctggcaaag
     2581 tgactggatg taccaggttc cctctgcagg gtttcggtag gaaacacagt tattcacagt
     2641 ggccatcccc agcagaaagt cagcctcatc cgggatatat ctcgtttgag gtgatgataa
     2701 atccatttct aaatagggtt gctcctctga atcagtctca acaggtatac ctttctggta
     2761 gttatccccc tgagcagcct gaataaaaaa cactttgggt tttccagcaa gggaagggca
     2821 cttcaaacca gtgaactgag atgtcagctc atagatgggg gcctcctgtc catcagtgcc
     2881 atagatgatg cccttgtctc catgggagag gatacagcag atgaagcagt ccatgttact
     2941 gtggtccatg agttggtaga ttttcaaaat ctcatagatt tgctctactg tgcagtcatc
     3001 gtggggcttg atctcaaaat gaagctcttc aaaggtcgtg gtcaaagccc ctgcatccaa
     3061 gtgtgttcca ttcctgtccc taatgctgtg aagtttgggc actttctccc gtgcttttgc
     3121 aaaattgtga ttgttgatga tcagacagta tccccgaggt ttgcttttca tttggtaaac
     3181 tttgtccaaa gtctgtgatt cactatcctg ttctcttgga gagtccgaga ttgtcatatg
     3241 ggatcctcta gaacctccac ctccactagt ttccagtttt agaagctcca catcgaagac
     3301 gagagtggca tgtggtggga tgatgcctgg gtgcccagtg gcaccatagg cataatctgg
     3361 agatatagtc agtttggctc tctgacccac actcatctgg gcaacccctt cttcccagcc
     3421 tcggatcacc tcctgcttgc ctagcataaa cttaaagggc ttgtttctgt cccgggagga
     3481 atcaactttc tttccatctt caagcatccc ggtgtagtgc accacgcagg tctggccgcg
     3541 cttggggaag gtgcgcccgt ctcctgggga gatagtctcc acctgcactc ctctagagcg
     3601 ctggctgggg tccttaggct tgctcttgct actccccatg gtagatccgg ccggcgccta
     3661 gagaaggagt gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa
     3721 gggggagaga cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct
     3781 tgacccaggg aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt
     3841 ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc
     3901 ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga
     3961 aggtaaccca acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat
     4021 ctttccagtg acctaaggtc aaacttaagg gagtggtaac agtctggccc taattttcag
     4081 acaaatacag aaacacagtc agacagagac aacacagaac gatgctgcag cgctgcagca
     4141 gacaagacgc gcggcttcgg ttccaaaccg aaagcaaaaa ttcagacgga ggcgggaact
     4201 gttttaggtt ctcgtctcct accagaacca catatcctga cggggtcgga ttccacatcg
     4261 actcccttcc tcaggtcggg ccacaaaaac ggcccccaaa gtccctggga cgtctcccag
     4321 ggttgcggcc gggtgttcag aactcgtcag ttccaccacg ggtccgccag atacagagct
     4381 agttagctaa ctagtaccga cgcaggcgca taaaatcagt catagacact agacaatcgg
     4441 acagacacag ataagttgct ggccagcttg cctcccggtg gtgggtcggt ggtccctggg
     4501 caggggtctc ccgatcccgg acgagccccc aaatgaaaga cccccgctga cgggtagtca
     4561 atcactcaga ggagaccctc ccaaggaaca gcgagaccac aagtcggatg caactgcaag
     4621 agggtttatt ggatacacgg gtacccgggc gactcagtca atcggaggac tggcgccccg
     4681 agtgaggggt tgtgggctct tttattgagc tcggggagca gaagcgcgcg aacagaagcg
     4741 agaagcgaac tgattggtta gttcaaataa ggcacagggt catttcaggt ccttggggca
     4801 ccctggaaac atctgatggt tctctagaaa ctgctgaggg ctggaccgca tctggggacc
     4861 atctgttctt ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc
     4921 atattcagct gttccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga
     4981 tatcctgttt ggcccatatt caggctgcag gtggcacttt tcggggaaat gtgcgcggaa
     5041 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5101 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5161 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5221 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5281 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5341 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5401 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5461 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5521 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5581 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5641 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5701 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5761 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5821 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5881 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5941 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6001 tgtcagacca agtttactca tatatacttt agattgattt gcggccgcaa aaggatctag
     6061 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6121 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6181 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6241 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6301 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6361 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6421 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6481 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6541 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6601 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6661 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6721 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6781 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6841 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6901 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6961 cgttggccga ttcattaatg caactatggc catttaatgt aaatacttaa gaaaaaaaac
     7021 caaattaatt ttgatacatg ctgcatgtga aagacccccg ctgacgggta gtcaatc
//
