LOCUS       V004057                 9018 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004057
VERSION     V004057
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9018)
  AUTHORS   Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Jian-Yu V, Chao
            SS, Hsu A, Pham V, Naghavian L, Dozier LE, Patrick G, Coleman T.
  TITLE     Biosynthesis of Orthogonal Molecules Using Ferredoxin and
            Ferredoxin-NADP+ Reductase Systems Enables Genetically Encoded PhyB
            Optogenetics
  JOURNAL   ACS Synth Biol (2018) In press
   PUBMED   29301067
REFERENCE   2  (bases 1 to 9018)
  AUTHORS   Kyriakakis P.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-NOV-2017) Bioengineering, UCSD, 9500 Gilman drive,
            PFBH room 251 MC0412, La Jolla, CA 92093, United States of America
REFERENCE   3  (bases 1 to 9018)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9018)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
            Biol (2018) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-NOV-2017) Bioengineering, UCSD, 9500 Gilman drive, PFBH room 251
            MC0412, La Jolla, CA 92093, United States of America"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1264..1971
                     /label="mOrange"
                     /note="orange monomeric derivative of DsRed fluorescent
                     protein (Shanet et al., 2004)"
     misc_feature    1972..2037
                     /label="P2A peptide"
                     /note="P2A peptide"
     CDS             1981..2037
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /label="P2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    2038..2124
                     /label="transit peptide MTS"
                     /note="transit peptide MTS"
     CDS             2038..2112
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label="COX8 presequence"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2140..2157
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             2170..2883
                     /label="superfolder GFP"
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     promoter        2903..3232
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3242..3838
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             4134..4367
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4524..4748
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      4791..4800
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        4844..4948
                     /label="AmpR promoter"
     CDS             4949..5806
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5980..6568
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     LTR             6727..7360
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7407..7532
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8025..8258
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8443..8487
                     /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             8636..8677
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    8774..8891
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        9018
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattgatcc ttcgaactag
     1201 aattcgaagc ttgagctcga gatctgagtc cggtagcgct agttcgaagg atccgccgcc
     1261 accatggtga gcaagggcga ggagaataac atggccatca tcaaggagtt catgcgcttc
     1321 aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1381 ggccgcccct acgagggctt tcagaccgct aagctgaagg tgaccaaggg tggccccctg
     1441 cccttcgcct gggacatcct gtcccctcag ttcacctacg gctccaaggc ctacgtgaag
     1501 caccccgccg acatccccga ctacttcaag ctgtccttcc ccgagggctt caagtgggag
     1561 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1621 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1681 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1741 ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg ccactacacc
     1801 tccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacatc
     1861 gtcggcatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1921 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa aggaagcgga
     1981 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctatg
     2041 agtgttctga ccccattgct gttgagagga ctgactggtt ccgccaggag gttgcccgtg
     2101 ccaagagcca aaatacatag cttagctagc ttaattaaac atcatcacca ccaccattct
     2161 agactcgaga tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa
     2221 ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca
     2281 actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc ggtaccttgg
     2341 ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat
     2401 atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg
     2461 atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat
     2521 accctggtaa accgcattga gctgaaaggc attgacttta aagaagacgg caatatcctg
     2581 ggccataagc tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa
     2641 aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag
     2701 ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac
     2761 aatcactatc tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat
     2821 atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac
     2881 aaatgagtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2941 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3001 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3061 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3121 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3181 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac
     3241 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     3301 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     3361 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     3421 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     3481 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     3541 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     3601 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     3661 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     3721 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     3781 cgacgtcgag tgcccgaagg accgcgcgac ctggtgcatg acccgcaagc ccggtgcctg
     3841 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat
     3901 aaaataaaag attttattta gtctccagaa aaagggggcg gccgctcgag acctagaaaa
     3961 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     4021 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     4081 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     4141 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     4201 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     4261 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     4321 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     4381 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc
     4441 ttgacattat aatagattta gcaggaattg aactaggagt ggagcacaca ggcaaagttc
     4501 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4561 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4621 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4681 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4741 ctctatggct tctgaggcgg aaagaaccag ctgggggcgc gcccctcgag gccgccatgg
     4801 tcatagctgt ttgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4861 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4921 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4981 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5041 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5101 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5161 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5221 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5281 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5341 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5401 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5461 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5521 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5581 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5641 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5701 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5761 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5821 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5881 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5941 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6001 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6061 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6121 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6361 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6601 ggccttttgc cggaccgcca ctgccaatta cctgtggttt catttactct aaacctgtga
     6661 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
     6721 agtagttgga agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct
     6781 accacacaca aggctacttc cctgattagc agaactacac accagggcca ggggtcagat
     6841 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag
     6901 aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg
     6961 acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg
     7021 cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac
     7081 tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc
     7141 tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag
     7201 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     7261 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     7321 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
     7381 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
     7441 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
     7501 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     7561 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     7621 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     7681 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     7741 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     7801 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     7861 cacagcaacc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     7921 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     7981 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     8041 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     8101 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     8161 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     8221 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     8281 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     8341 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     8401 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     8461 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     8521 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     8581 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     8641 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     8701 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcca caaatggcag
     8761 tattcatcca caattttaaa agaaaggggg ggattggggg gtacagtgca ggggaaagaa
     8821 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     8881 ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggtcg ataagctttg
     8941 caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc
     9001 ttgaaaggag tgggaatt
//
