LOCUS       V001414                 9979 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001414
VERSION     V001414
KEYWORDS    pHR_EGFPligand
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9979)
  AUTHORS   Morsut L, Roybal KT, Xiong X, Gordley RM, Coyle SM, Thomson M, Lim
            WA
  TITLE     Engineering Customized Cell Sensing and Response Behaviors Using
            Synthetic Notch Receptors.
  JOURNAL   Cell. 2016 Feb 11;164(4):780-91. doi: 10.1016/j.cell.2016.01.012.
            Epub 2016 Jan 28.
   PUBMED   26830878
REFERENCE   2  (bases 1 to 9979)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9979)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi:
            "10.1016/j.cell.2016.01.012"; journalName: "Cell"; date:
            "2016-02-11- 11"; volume: "164"; issue: "4"; pages: "780-91"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          655..720
                     /label="small t intron"
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             850..870
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    1295..1429
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     LTR             1597..2230
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2277..2402
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    2899..3132
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             3317..3361
                     /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             3510..3551
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    3646..3763
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        3916..4323
                     /label="SFFV promoter"
                     /note="spleen focus-forming virus long terminal repeat
                     (LTR) promoter"
     regulatory      4391..4400
                     /label="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4460..5176
                     /label="EGFP"
                     /note="enhanced GFP"
     misc_feature    5439..6027
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(6030..6046)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(6031..6047)
                     /label="pBluescriptKS"
                     /note="For pBluescript vector"
     LTR             6137..6370
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(6395..6413)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6703..6722)
                     /label="pBRrevBam"
                     /note="pBR322 vectors, tet region, downstream of BamHI,
                     reverse primer"
     primer_bind     complement(6936..6955)
                     /label="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     7055..7077
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7115..7133)
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        7201..7305
                     /label="AmpR promoter"
     CDS             7306..8163
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      8337..8925
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     9079..9096
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     promoter        9171..9500
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 tggtactgcg gttgcgattc gtgaaatgta tccaaaagcg cactttgtca ccatcttcgc
       61 aaaaccggct ggtcgtccgc tggttgatga ctatgttgtt gatatcccgc aagatacctg
      121 gattgaacag ccgtgggata tgggcgtcgt attcgtcccg ccaatctccg gtcgctaatc
      181 ttttcaacgc ctggcactgc cgggcgttgt tctttttaac ttcaggcggg ttacaatagt
      241 ttccagtaag tattctggag gctgcatcca tgacacaggc aaacctgagc gaaaccctgt
      301 tcaaaccccg ctttaaacat cctgaaacct cgacgctagt ccgccgcttt aatcacggcg
      361 cacaaccgcc tgtgcagtcg gcccttgatg gtaaaaccat ccctcactgg tatcgcatga
      421 ttaaccgtct gatgtggatc tggcgcggca ttgacccacg cgaaatcctc gacgtccagg
      481 cacgtattgt gatgagcgat gccgaacgta ccgacgatga tttatacgat acggtgattg
      541 gctaccgtgg cggcaactgg atttatgagt gggccccgga tctttgtgaa ggaaccttac
      601 ttctgtggtg tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat
      661 ataaaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag
      721 attccaacct atggaactga tgaatgggag cagtggtgga atgcctttaa tgaggaaaac
      781 ctgttttgct cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat
      841 tctactcctc caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg
      901 ctaagttttt tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt tgctatttac
      961 accacaaagg aaaaagctgc actgctatac aagaaaatta tggaaaaata ttctgtaacc
     1021 tttataagta ggcataacag ttataatcat aacatactgt tttttcttac tccacacagg
     1081 catagagtgt ctgctattaa taactatgct caaaaattgt gtacctttag ctttttaatt
     1141 tgtaaagggg ttaataagga atatttgatg tatagtgcct tgactagaga tcataatcag
     1201 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     1261 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     1321 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1381 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac
     1441 tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta
     1501 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa
     1561 ttgtatttgt taaatatgta ctacaaactt agtagttgga agggctaatt cactcccaaa
     1621 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc
     1681 agaactacac accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc
     1741 tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt
     1801 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt
     1861 ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga
     1921 actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg
     1981 gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt
     2041 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     2101 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     2161 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     2221 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     2281 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     2341 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     2401 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     2461 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     2521 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     2581 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     2641 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     2701 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag
     2761 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     2821 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     2881 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     2941 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     3001 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     3061 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     3121 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     3181 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     3241 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     3301 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     3361 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     3421 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     3481 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     3541 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     3601 gaacggatct cgacggtatc gccaaatggc agtattcatc cacaatttta aaagaaaagg
     3661 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     3721 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     3781 cagcagagat ccagtttgga tcgataagct tgatatcgaa ttcctgcagc cccgataaaa
     3841 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     3901 ggcaagctag ctgcagtaac gccattttgc aaggcatgga aaaataccaa accaagaata
     3961 gagaagttca gatcaagggc gggtacatga aaatagctaa cgttgggcca aacaggatat
     4021 ctgcggtgag cagtttcggc cccggcccgg ggccaagaac agatggtcac cgcagtttcg
     4081 gccccggccc gaggccaaga acagatggtc cccagatatg gcccaaccct cagcagtttc
     4141 ttaagaccca tcagatgttt ccaggctccc ccaaggacct gaaatgaccc tgcgccttat
     4201 ttgaattaac caatcagcct gcttctcgct tctgttcgcg cgcttctgct tcccgagctc
     4261 tataaaagag ctcacaaccc ctcactcggc gcgccagtcc tccgacagac tgagtcgccc
     4321 gggggggatc tggagctctc gagaattctc acgcgtcaag tggagcaagg caggtggaca
     4381 gtggatcgcc gccaccatgg agaccgacac cctcttgctc tgggtcctgc tgctgtgggt
     4441 tccgggtagt accggagaca tggtttctaa gggtgaagaa ttgttcaccg gagttgtgcc
     4501 catcctggtt gagctggacg gagacgttaa cggacacaag ttctcagtga gcggcgaagg
     4561 agagggcgac gctacgtacg gtaaactgac gttgaagttc atctgtacaa caggaaagct
     4621 gccagtgccc tggcctacac tcgtgaccac cctcacctac ggagtccagt gcttctcccg
     4681 gtaccccgac cacatgaaac agcacgattt cttcaagtcc gcgatgcctg agggctatgt
     4741 tcaggaacgc accatattct tcaaggacga cggtaactat aaaaccaggg ctgaagttaa
     4801 gttcgagggg gacacgttgg taaatagaat tgagttgaaa ggaatcgatt tcaaagaaga
     4861 cggcaatatt ttgggccaca agctggagta caactacaac agccacaacg tgtatattat
     4921 ggctgataag cagaagaacg gcatcaaggt gaacttcaaa atccgacaca atatcgagga
     4981 tggctctgtg caactggccg accattacca gcaaaacacg cctatcggcg acggtccggt
     5041 tttgctgcca gataaccact acctctccac gcaatcagcc ctctccaagg atcctaacga
     5101 aaaaagagac catatggttc tgcttgaatt cgtgacggct gctgggatca ccctgggaat
     5161 ggatgagctg tataagaacg ccgtgggcca ggatacgcag gaggtgatcg tcgtgcccca
     5221 cagcttgccc ttcaaggtcg tggttattag tgctatcctg gcgttggtgg tgctgactat
     5281 tatctccctt atcatactta tcatgctgtg gcaaaaaaaa ccccgataag atccttgact
     5341 tgcggccgca actcccacct gcaacatgcg tgactgactg aggccgcgac tctagagtcg
     5401 acctgcaggc atgcaagctt gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     5461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     5521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     5581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     5641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     5701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     5761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     5821 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     5881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     5941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     6001 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgaggg aattaattcg
     6061 agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6121 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     6181 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6241 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6301 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6361 atctctagca gcatctagaa ttaattccgt gtattctata gtgtcaccta aatcgtatgt
     6421 gtatgataca taaggttatg tattaattgt agccgcgttc taacgacaat atgtacaagc
     6481 ctaattgtgt agcatctggc ttactgaagc agaccctatc atctctctcg taaactgccg
     6541 tcagagtcgg tttggttgga cgaaccttct gagtttctgg taacgccgtc ccgcacccgg
     6601 aaatggtcag cgaaccaatc agcagggtca tcgctagcca gatcctctac gccggacgca
     6661 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     6721 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     6781 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     6841 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
     6901 cgcataaggg agagcgtcga atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6961 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     7021 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     7081 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     7141 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     7201 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     7261 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     7321 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7381 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     7441 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     7501 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7561 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     7621 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7681 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     7741 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7801 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     7861 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7921 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7981 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     8041 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     8101 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     8161 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     8221 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     8281 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     8341 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     8401 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     8461 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     8521 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     8581 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     8641 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     8701 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     8761 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     8821 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8881 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8941 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     9001 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     9061 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     9121 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctgtggaat gtgtgtcagt
     9181 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     9241 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     9301 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     9361 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     9421 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     9481 gaggcctagg cttttgcaaa aagcttggac acaagacagg cttgcgagat atgtttgaga
     9541 ataccacttt atcccgcgtc agggagaggc agtgcgtaaa aagacgcgga ctcatgtgaa
     9601 atactggttt ttagtgcgcc agatctctat aatctcgcgc aacctatttt cccctcgaac
     9661 actttttaag ccgtagataa acaggctggg acacttcaca tgagcgaaaa atacatcgtc
     9721 acctgggaca tgttgcagat ccatgcacgt aaactcgcaa gccgactgat gccttctgaa
     9781 caatggaaag gcattattgc cgtaagccgt ggcggtctgt accgggtgcg ttactggcgc
     9841 gtgaactggg tattcgtcat gtcgataccg tttgtatttc cagctacgat cacgacaacc
     9901 agcgcgagct taaagtgctg aaacgcgcag aaggcgatgg cgaaggcttc atcgttattg
     9961 atgacctggt ggataccgg
//
