LOCUS       V004068                 8593 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004068
VERSION     V004068
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8593)
  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 8593)
  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 8593)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8593)
  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..8593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        655..673
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             693..2264
                     /gene="PIF3"
                     /label="Transcription factor PIF3"
                     /note="Transcription factor PIF3 from Arabidopsis thaliana.
                     Accession#: O80536"
     CDS             2328..2477
                     /label="VP64"
                     /note="tetrameric repeat of the minimal activation domain
                     of herpes simplex virus VP16 (Beerli et al., 1998)"
     CDS             2502..2522
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             2952..3308
                     /label="RelA (p65) AD"
                     /note="transcriptional activation domain of human RelA,
                     also known as p65 (O'Shea and Perkins, 2008)"
     CDS             3327..3896
                     /label="Rta AD"
                     /note="transcriptional activation domain from the human
                     herpesvirus 4 (Epstein-Barr virus) replication and
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992;
                     Chavez et al., 2015)"
     promoter        complement(3937..3955)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3981..4205
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4251..4679
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4693..5023
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5090..5881
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    6058..6191
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6228..6244)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6252..6268)
                     /label="lac operator"
                     /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(6276..6306)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6321..6342)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6630..7218)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7392..8249)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8250..8354)
                     /label="AmpR promoter"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg actatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagaccc aagcttggta ccatgcctct gtttgagctt ttcaggctca
      721 ccaaagctaa gcttgaatct gctcaagaca ggaacccttc tccacctgta gatgaagttg
      781 tggagctggt gtgggaaaat ggtcagatat caactcaaag tcagtcaagt agatcgagga
      841 acattcctcc accacaagca aactcttcta gagctagaga gattggaaat ggctcaaaga
      901 cgactatggt ggacgagatc cctatgtcag tgccatcact aatgacgggt ttgagtcaag
      961 acgatgactt tgttccatgg ttgaatcatc atccctccct tgatggatat tgctctgatt
     1021 tcttgcgtga tgtgtcgtct cctgttactg tcaacgagca agagagtgat atggcggtaa
     1081 accaaactgc tttcccgttg tttcagagaa gaaaggatgg caatgaatca gctcctgctg
     1141 cttcttcgtc gcagtataac ggtttccaat cgcattctct gtatggaagt gatagagcta
     1201 gagatcttcc tagccaacaa accaatccgg atcggtttac tcagacgcag gaaccactaa
     1261 ttactagtaa caagcctagt ttggtcaact tttcacattt cttacgccct gcaacttttg
     1321 cgaagactac taataataac cttcatgaca ctaaagaaaa gagtcctcaa agcccgccaa
     1381 atgtgtttca gaccagagtt cttggagcta aagactctga agataaggtt cttaacgagt
     1441 ctgttgcttc tgctacgcct aaagataacc aaaaggcttg cctaatatca gaggactcat
     1501 gtagaaaaga ccaagagagt gaaaaagcag ttgtatgttc ttctgttggc tcgggtaata
     1561 gtctcgatgg cccatccgaa agtccttcac tttctttaaa gagaaagcat tcgaatattc
     1621 aagacattga ctgtcatagt gaagatgtgg aagaagaatc aggagatgga agaaaggaag
     1681 caggtccatc tcgaacgggt ttgggttcaa agagaagccg ctctgcagaa gtgcataatc
     1741 tgtctgaaag gagacggcgt gataggatca acgagaagat gcgtgccctg caagaactca
     1801 ttccaaactg taacaaggtg gacaaagctt cgatgctaga tgaagccatc gagtatctca
     1861 agtcactcca acttcaagtg cagatcatgt caatggcgtc tggttactat ctgccaccgg
     1921 cggttatgtt cccaccgggt atggggcatt acccggcagc agctgctgca atggcaatgg
     1981 gtatgggaat gccttatgca atgggcttgc ctgatttgag ccgtggtggt tcatcggtta
     2041 accacggacc acagttccaa gtctcgggga tgcaacaaca accagtggcg atgggtattc
     2101 cacgtgtctc tggtggtggt atctttgccg gttcttcgac gattggcaat ggctcgacta
     2161 gagatttatc tggttctaaa gatcaaacaa cgacgaataa caacagtaac ttgaaaccaa
     2221 taaagagaaa acaggggtct tctgatcagt tttgtggatc gtcggacagc gccggcagcg
     2281 ccggcagcgc cggcagcgga tccgaggcta gtggatctgg gagagcagat gctctggatg
     2341 actttgatct ggacatgctg ggctccgacg cactcgatga cttcgatctg gatatgcttg
     2401 gcagcgacgc actggacgac ttcgatctgg acatgttggg atctgacgcg ctggacgact
     2461 tcgatctgga catgctgatc aattctcgct catccggttc accaaagaaa aagaggaagg
     2521 ttgggagtca gtatcttccc gataccgacg atcggcaccg gatagaggaa aagaggaagc
     2581 ggacatacga gaccttcaaa tctatcatga agaagtcacc cttctctgga cccaccgatc
     2641 ccagaccccc gccccggcgc attgccgtac cctcccgcag tagcgcctca gtccccaagc
     2701 ctgcacctca accttatcct tttaccagtt cactgtctac cattaattac gacgagtttc
     2761 cgacgatggt gtttccctct ggccagatat ctcaggcctc tgccctggcg cctgcacctc
     2821 cacaggtact gcctcaggca ccggcacccg ccccggctcc tgcaatggtg tccgccctcg
     2881 ctcaggcccc cgcacctgtg cctgtacttg cccctggccc cccgcaggct gtcgctcctc
     2941 ctgcccccaa gcctacccag gccggggagg gaactctgtc agaggccttg ttgcagttgc
     3001 agttcgatga cgaggacctg ggagctcttc tgggcaattc taccgacccc gccgtcttca
     3061 ctgacctggc aagcgtggac aatagcgaat ttcaacagtt gcttaaccag ggtattccgg
     3121 ttgctccgca tactactgag cccatgctga tggaataccc tgaggccatc accagattgg
     3181 tcaccggggc acagcggcct ccggacccag ctcccgcccc tctgggcgcc cccggactgc
     3241 caaacggcct gctgtccgga gacgaagatt tctctagtat tgccgacatg gacttttctg
     3301 cccttttggg cagcgggagc ggttccagag atagtcgcga aggaatgttc ctgccaaaac
     3361 ccgaagcagg cagcgccata agtgacgtgt tcgagggacg ggaagtgtgc cagccaaaga
     3421 ggattagacc attccaccca ccaggctctc cctgggctaa taggcccctc cctgcttcat
     3481 tggcgcctac tccgactggt cccgtccacg agccagtggg gtctctgact cctgcacctg
     3541 tgccacagcc cctggaccca gccccagctg tcacccccga ggccagccac ctgctcgaag
     3601 atccagatga ggaaacctcc caggccgtaa aggccttgcg ggaaatggcc gatactgtaa
     3661 tcccacagaa agaggaggcc gccatttgtg ggcaaatgga tctcagtcat ccccctccaa
     3721 ggggtcacct ggatgaattg accacaacct tggaatcaat gacagaagac ctcaatctgg
     3781 atagccccct gacgccggag ctgaacgaga ttcttgacac ttttctcaac gatgaatgcc
     3841 tcttgcatgc catgcatatc agcaccggcc tgtctatttt tgatacctcc ttgttttgat
     3901 aagcggccgc tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     3961 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4021 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4081 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4141 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4201 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4261 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4321 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4381 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     4441 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4501 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4561 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4621 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4681 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
     4741 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4801 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4861 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4921 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4981 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     5041 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5101 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5161 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5221 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     5281 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5341 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5401 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5461 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5521 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5581 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5641 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5701 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5761 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5821 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5881 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5941 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6001 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     6061 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6121 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6181 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     6241 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6301 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6361 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6421 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6481 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6541 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6601 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6661 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6721 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6781 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     6841 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6901 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6961 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7021 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     7081 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7141 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7201 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7261 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7321 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7381 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7441 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7501 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7561 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7621 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7681 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7741 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7801 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7861 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7921 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7981 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8041 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8101 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8161 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8221 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8281 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8341 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     8401 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     8461 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     8521 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     8581 ttgcgctgct tcg
//
