LOCUS       40924_34759        6550 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPKm-118, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6550)
  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 6550)
  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 6550)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6550)
  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..6550
                     /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      125..166
                     /label=pFR-luc promoter
                     /note="pFR-luc promoter"
                     /regulatory_class="promoter"
     CDS             207..1856
                     /label=luciferase
                     /note="firefly luciferase"
     promoter        complement(1894..1912)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    1938..2162
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2208..2636
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2650..2980
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3047..3838
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4015..4148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4185..4201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4209..4225)
                     /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(4233..4263)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4278..4299)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4587..5175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5349..6206)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6207..6311)
                     /label=AmpR promoter
ORIGIN
        1 cgaaagcttg catgcgagta ctgtcctccg agcggagtac tgtcctccga gcggagtact
       61 gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc
      121 ctccgagcgg agactctaga gggtatataa tggatccccg ggtaccgagc tcgaattcca
      181 gcttggcatt ccggtactgc cgccacatgg aagacgccaa aaacataaag aaaggcccgg
      241 cgccattcta tcctctagag gatggaaccg ctggagagca actgcataag gctatgaaga
      301 gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag gtgaacatca
      361 cgtacgcgga atacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc
      421 tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg
      481 tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac
      541 gtgaattgct caacagtatg aacatttcgc agcctaccgt agtgtttgtt tccaaaaagg
      601 ggttgcaaaa aattttgaac gtgcaaaaaa aattaccaat aatccagaaa attattatca
      661 tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc
      721 tacctcccgg ttttaatgaa tacgattttg taccagagtc ctttgatcgt gacaaaacaa
      781 ttgcactgat aatgaattcc tctggatcta ctgggttacc taagggtgtg gcccttccgc
      841 atagaactgc ctgcgtcaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca
      901 ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta
      961 cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc
     1021 tgtttttacg atcccttcag gattacaaaa ttcaaagtgc gttgctagta ccaaccctat
     1081 tttcattctt cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa
     1141 ttgcttctgg gggcgcacct ctttcgaaag aagtcgggga agcggttgca aaacgcttcc
     1201 atcttccagg gatacgacaa ggatatgggc tcactgagac tacatcagct attctgatta
     1261 cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga
     1321 aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcagagaggc gaattatgtg
     1381 tcagaggacc tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga
     1441 ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact
     1501 tcttcatagt tgaccgcttg aagtctttaa ttaaatacaa aggatatcag gtggcccccg
     1561 ctgaattgga atcgatattg ttacaacacc ccaacatctt cgacgcgggc gtggcaggtc
     1621 ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga
     1681 cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt
     1741 tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg
     1801 caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagtcc aaattgtaag
     1861 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     1921 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1981 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2041 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2101 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2161 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2221 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2281 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2341 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     2401 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2461 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2521 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     2581 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2641 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag
     2701 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2761 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2821 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2881 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2941 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3001 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3061 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3121 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3181 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3241 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3301 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3361 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3421 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3481 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3541 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3601 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3661 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3721 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3781 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3841 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3901 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3961 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4021 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4081 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4141 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4201 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4261 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4321 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4381 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4441 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4501 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     4801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5221 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5341 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5401 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5461 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5521 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5581 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5641 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5701 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5761 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5821 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5881 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5941 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6001 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6061 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6121 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6181 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6241 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6301 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     6361 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     6421 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     6481 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     6541 cgctgcttcg
//
