LOCUS       40924_34784        6643 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPKm-202, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6643)
  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 6643)
  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 6643)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6643)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2018) In press"
COMMENT     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"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    33..127
                     /label=5X UAS
                     /note="five tandem copies of the 'ScaI site' 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     regulatory      join(125..157,253..258)
                     /label=pFR-luc promoter
                     /note="pFR-luc promoter"
                     /regulatory_class="promoter"
     regulatory      158..252
                     /label=CMV min
                     /note="CMV min"
                     /regulatory_class="promoter"
     CDS             300..1949
                     /label=luciferase
                     /note="firefly luciferase"
     promoter        complement(1987..2005)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2031..2255
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2301..2729
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2743..3073
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3140..3931
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4108..4241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4278..4294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4302..4318)
                     /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(4326..4356)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4371..4392)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4680..5268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5442..6299)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6300..6404)
                     /label=AmpR promoter
ORIGIN
        1 cgaaagcttg catgcgagta ctgtcctccg agcggagtac tgtcctccga gcggagtact
       61 gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc
      121 ctccgagcgg agactctaga gggtatataa tggatcccca agcagagctc gtttagtgaa
      181 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
      241 ccgatccagc ctggtaccga gctcgaattc cagcttggca ttccggtact gccgccacca
      301 tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatcctcta gaggatggaa
      361 ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg
      421 cttttacaga tgcacatatc gaggtgaaca tcacgtacgc ggaatacttc gaaatgtccg
      481 ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat
      541 gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg
      601 cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgaacattt
      661 cgcagcctac cgtagtgttt gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa
      721 aaaaattacc aataatccag aaaattatta tcatggattc taaaacggat taccagggat
      781 ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt
      841 ttgtaccaga gtcctttgat cgtgacaaaa caattgcact gataatgaat tcctctggat
      901 ctactgggtt acctaagggt gtggcccttc cgcatagaac tgcctgcgtc agattctcgc
      961 atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg
     1021 ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc
     1081 gagtcgtctt aatgtataga tttgaagaag agctgttttt acgatccctt caggattaca
     1141 aaattcaaag tgcgttgcta gtaccaaccc tattttcatt cttcgccaaa agcactctga
     1201 ttgacaaata cgatttatct aatttacacg aaattgcttc tgggggcgca cctctttcga
     1261 aagaagtcgg ggaagcggtt gcaaaacgct tccatcttcc agggatacga caaggatatg
     1321 ggctcactga gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg
     1381 cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa
     1441 cgctgggcgt taatcagaga ggcgaattat gtgtcagagg acctatgatt atgtccggtt
     1501 atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg
     1561 gagacatagc ttactgggac gaagacgaac acttcttcat agttgaccgc ttgaagtctt
     1621 taattaaata caaaggatat caggtggccc ccgctgaatt ggaatcgata ttgttacaac
     1681 accccaacat cttcgacgcg ggcgtggcag gtcttcccga cgatgacgcc ggtgaacttc
     1741 ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt
     1801 acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg
     1861 aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa
     1921 aggccaagaa gggcggaaag tccaaattgt aagcggccgc tcgagcatgc atctagaggg
     1981 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     2041 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2101 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2161 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2221 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2281 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2341 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2401 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     2461 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2521 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2581 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2641 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     2701 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2761 ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2821 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2881 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2941 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3001 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3061 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3121 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3181 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3241 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3301 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3361 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3421 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3481 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3541 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3601 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3661 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3721 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3781 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3841 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3901 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3961 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4021 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4081 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4141 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4201 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4261 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4321 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4381 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4441 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4501 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4561 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4621 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4681 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4741 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4801 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4861 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4921 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4981 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5041 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5101 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5161 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5521 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5581 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5641 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5701 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5761 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5821 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5881 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5941 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6001 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6061 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6121 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6181 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6241 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6301 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6361 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6421 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta
     6481 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     6541 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     6601 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcg
//
