LOCUS       V016574                 8785 bp    DNA     circular SYN 18-MAY-2019
DEFINITION  Exported.
ACCESSION   V016574
VERSION     V016574
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8785)
  AUTHORS   Chung HK, Zou X, Bajar BT, Brand VR, Huo Y, Alcudia JF, Ferrell JE
            Jr., Lin MZ.
  TITLE     A compact synthetic pathway rewires cancer signaling to therapeutic
            effector release
  JOURNAL   Science 364 (6439) (2019)
   PUBMED   31048459
REFERENCE   2  (bases 1 to 8785)
  AUTHORS   Chung HK, Lin MZ.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-APR-2019) Bioengineering, Stanford University, 269
            Campus Drive, CCRS2100, Stanford, CA 94305, United States
REFERENCE   3  (bases 1 to 8785)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Science 364
            (6439) (2019)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (16-APR-2019) Bioengineering, Stanford University, 269 Campus Drive,
            CCRS2100, Stanford, CA 94305, United States"
FEATURES             Location/Qualifiers
     source          1..8785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    12..564
                     /label="nonstandard type: promoter; CMV_imm early promoter
                     2"
     misc_feature    67..354
                     /label="CAG enhancer"
                     /note="CAG enhancer"
     misc_feature    521..541
                     /label="nonstandard type: primer_bind; CMV fwd primer"
     LTR             584..764
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    811..936
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1435..1668
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1853..1897
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             2046..2087
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2195..2312
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2371..2684
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das
                     et al., 1988)"
     primer_bind     2701..2717
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(2759..2792)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        2908..3211
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3212..3415
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     3414..3465
                     /label="p1317"
                     /note="p1317"
     misc_feature    3447..4004
                     /label="Shc-PTB"
                     /note="Shc-PTB"
     misc_feature    3810..3830
                     /label="HIF1a"
                     /note="HIF1a"
     CDS             4008..4031
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    4068..4646
                     /label="nonstandard type: Protease; NS3Tprotease"
     misc_feature    4227..4229
                     /label="nonstandard type: misc; T54"
     primer_bind     4394..4426
                     /label="p1278"
                     /note="p1278"
     misc_feature    complement(4412..4442)
                     /direction=LEFT
                     /label="nonstandard type: primer_bind_reverse; p1502 MreI
                     rev"
     misc_feature    4647..4712
                     /label="P2A"
                     /note="P2A"
     misc_feature    complement(4653..4712)
                     /direction=LEFT
                     /label="nonstandard type: primer_bind_reverse; p1401 P2A
                     reverse"
     CDS             4713..5363
                     /label="mKO2"
                     /note="monomeric Kusabira-Orange 2 fluorescent protein"
     misc_feature    5376..5417
                     /label="nonstandard type: epitope; V5"
     misc_feature    5418..5450
                     /label="nonstandard type: Substrate; substrate 4"
     misc_feature    complement(5451..5489)
                     /direction=LEFT
                     /label="nonstandard type: primer_bind_reverse; p1514 GGS
                     linker"
     misc_feature    5469..5852
                     /label="nonstandard type: gBLOCK; HKC gBLOCK-5 VAV1"
     misc_feature    5490..5822
                     /label="nonstandard type: SH2; VAV1"
     primer_bind     5823..5858
                     /label="p1511 GGs linker before CAAX"
                     /note="p1511 GGs linker before CAAX"
     misc_feature    5880..5939
                     /label="CAAX"
                     /note="CAAX"
     misc_feature    5943..5948
                     /label="EcoRV"
                     /note="EcoRV"
     misc_feature    5961..6549
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(6552..6568)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    6734..6755
                     /label="U3PPT"
                     /note="U3PPT"
     misc_feature    6734..6749
                     /label="cPPT(1)"
                     /note="cPPT(1)"
     misc_feature    6791..6971
                     /label="truncHIV-1 3 LTR(1)"
                     /note="truncHIV-1 3 LTR(1)"
     rep_origin      complement(7033..7621)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7795..8652)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8653..8757)
                     /label="AmpR promoter"
ORIGIN
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga
      601 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      661 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      721 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
      781 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
      841 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
      901 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
      961 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     1021 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     1081 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     1141 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     1201 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     1261 accgcacagc aagcggccgg ccgcgctgat cttcagacct ggaggaggag atatgaggga
     1321 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1381 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1441 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1501 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1561 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1621 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1681 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1741 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1801 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1861 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1921 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1981 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     2041 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     2101 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc
     2161 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     2221 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2281 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     2341 cagtttggtt agtaccgggc ccgctctaga gatccgacgc cgccatctct aggcccgcgc
     2401 cggccccctc gcacagactt gtgggagaag ctcggctact cccctgcccc ggttaatttg
     2461 catataatat ttcctagtaa ctatagaggc ttaatgtgcg ataaaagaca gataatctgt
     2521 tctttttaat actagctaca ttttacatga taggcttgga tttctataag agatacaaat
     2581 actaaattat tattttaaaa aacagcacaa aaggaaactc accctaactg taaagtaatt
     2641 gtgtgttttg agactataaa tatcccttgg agaaaagcct tgttaacgcg cggtgaccct
     2701 cgaggtcgac ggtatcgata agctcgcttc acgagattcc agcaggtcga gggacctaat
     2761 aacttcgtat agcatacatt atacgaagtt atattaaggg ttccaagctt aagcggccgc
     2821 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     2881 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2941 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     3001 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     3061 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3121 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3181 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     3241 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     3301 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     3361 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     3421 agtgaaccgt cagatccgct agcaccatgg gcaagccact gcatcccaac gacaaagtca
     3481 tgggacccgg ggtttcctac ttggttcggt acatgggttg tgtggaggtc ctccagtcaa
     3541 tgcgtgccct ggacttcaac acccggactc aggtcaccag ggaggccatc agtctggtgt
     3601 gtgaggctgt gccgggtgct aagggggcga caaggaggag aaagccctgt agccgcccgc
     3661 tcagctctat cctggggagg agtaacctga aatttgctgg aatgccaatc actctcaccg
     3721 tctccaccag cagcctcaac ctcatggccg cagactgcaa acagatcatc gccaaccacc
     3781 acatgcaatc tatctcattt gcatccggca tgttagctcc ctatatccca gagtatgtcg
     3841 cctatgttgc caaagaccct gtgaatcaga gagcctgcca cattctggag tgtcccgaag
     3901 ggcttgccca ggatgtcatc agcaccattg gccaggcctt cgagttgcgc ttcaaacaat
     3961 acctcaggga catcgagcag gtgccccagc agcccaccct taagatggat tacaaggatg
     4021 acgacgataa gggctcttcc gggacaggct ccggatccgg cactagtgcg cccatcacgg
     4081 cgtacgccca gcagacgaga ggcctcctag ggtgtataat caccagcctg actggccggg
     4141 acaaaaacca agtggagggt gaggtccaga tcgtgtcaac tgctacccaa accttcctgg
     4201 caacgtgcat caatggggta tgctggactg tctaccacgg ggccggaacg aggaccatcg
     4261 catcacccaa gggtcctgtc atccagatgt ataccaatgt ggaccaagac cttgtgggct
     4321 ggcccgctcc tcaaggttcc cgctcattga caccctgtac ctgcggctcc tcggaccttt
     4381 acctggtcac gaggcacgcc gatgtcattc ccgtgcgccg gcgaggtgat agcaggggta
     4441 gcctgctttc gccccggccc atttcctact tgaaaggctc ctcggggggt ccgctgttgt
     4501 gccccgcggg acacgccgtg ggcctattca gggccgcggt gtgcacccgt ggagtggcta
     4561 aagcggtgga ctttatccct gtggagaacc tagagacaac catgagatcc ccggtgttct
     4621 cggacaattc ctccccacca gcggtgggct ctggagccac caacttcagc ctgctgaagc
     4681 aggcaggcga cgtggaagag aaccctggcc ctatggtgag cgtgatcaag cccgagatga
     4741 agatgaggta ctacatggac ggctccgtca atgggcatga gttcacaatc gagggtgagg
     4801 gcacaggcag accttacgag ggacatcagg agatgacact gcgcgtcaca atggccgagg
     4861 gcgggccaat gcctttcgcc ttcgacctgg tgtcccacgt gttctgttac ggccacagag
     4921 tttttaccaa gtacccagaa gagatcccag actatttcaa gcaggccttt cctgagggcc
     4981 tgtcctggga gaggtccctg gagttcgagg acggcggctc cgcctccgtg agcgcccaca
     5041 tcagcctgag gggcaacacc ttctaccaca agtccaagtt caccggcgtg aacttccccg
     5101 ccgacggccc catcatgcag aaccagagcg tggactggga gccctccacc gagaagatca
     5161 ccgccagcga cggcgtgctg aagggcgacg tgaccatgta cctgaagctg gagggcggcg
     5221 gcaaccacaa gtgccagatg aagaccacct acaaggccgc caaggagatc ctggagatgc
     5281 ccggcgacca ctacatcggc cacaggctgg tgaggaagac cgagggcaac atcaccgagc
     5341 aggtggagga cgccgtggcc caccttaagg gatccggtaa accaatcccc aatcctctgc
     5401 tgggtcttga tagcaccgat gagatggaag agtgctctca gcacctcccc ggcggttcgg
     5461 gaggttctgg tgggtcaggt ggcagtggcg gccctcctca ggacctgtct gttcatctct
     5521 ggtacgcagg ccccatggag cgggcagggg cagagagcat cctggccaac cgctcggacg
     5581 ggactttctt ggtgcggcag agggtgaagg atgcagcaga atttgccatc agcattaaat
     5641 ataacgtcga ggtcaagcac attaaaatca tgacagcaga aggactgtac cggatcacag
     5701 agaaaaaggc tttccggggg cttacggagc tggtggagtt ttaccagcag aactctctaa
     5761 aggattgctt caagtctctg gacaccacct tgcagttccc cttcaaggag cctgaaaaga
     5821 gaggatctag agggggatct ggtggatcag ggggtagcgg gggttctggc ggacaattca
     5881 agatgagcaa ggacggcaag aagaaaaaga agaagtccaa gactaagtgc gtgatcatgt
     5941 gagatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     6001 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     6061 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     6121 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     6181 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     6241 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     6301 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     6361 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     6421 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6481 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     6541 cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg agcaatcaca
     6601 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     6661 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
     6721 gacgcgtgat tttaaaagaa aaggggggac tggaagggct aattcactcc cactgctttt
     6781 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     6841 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     6901 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     6961 aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7021 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7081 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7141 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7201 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7261 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7321 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7381 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7441 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7501 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7561 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7621 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7681 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7741 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7801 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7861 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7921 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7981 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8041 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8101 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8161 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8221 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8281 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8341 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8401 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8461 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8521 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8581 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8641 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8701 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8761 atttccccga aaagtgccac ctgac
//
