LOCUS       40924_33492        8102 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNR.CFH, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8102)
  AUTHORS   Jackson HO, Bibby TS, Moore MC, Terry MJ.
  TITLE     Characterization of constitutive and inducible promoters in 
            Nannochloropsis gaditana (unpublished)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8102)
  AUTHORS   Jackson HO, Bibby TS, Moore MC, Terry MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-FEB-2017) Ocean and Earth Sciences, National 
            Oceanography Centre, Southampton, European Way, Southampton SO14 
            3ZH, England
REFERENCE   3  (bases 1 to 8102)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8102)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-FEB-2017) Ocean and Earth Sciences, National Oceanography 
            Centre, Southampton, European Way, Southampton SO14 3ZH, England"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..1319
                     /label=NR promoter
                     /note="NR promoter"
                     /regulatory_class="promoter"
     misc_feature    1323..1330
                     /label=NotI restriction site
                     /note="NotI restriction site"
     misc_feature    1342..1349
                     /label=AscI restriction site
                     /note="AscI restriction site"
     CDS             1356..1379
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             1386..1409
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1419..1445
                     /label=9xHis
                     /note="9xHis affinity tag"
     regulatory      1452..1959
                     /label=ATPa terminator
                     /note="ATPa terminator"
                     /regulatory_class="terminator"
     regulatory      1990..2682
                     /label=UEP promoter
                     /note="UEP promoter"
                     /regulatory_class="promoter"
     CDS             2689..3708
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     terminator      3743..3995
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4029..4045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4053..4069)
                     /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(4077..4107)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4122..4143)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      4184..4772
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4878..5860
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(6007..6807)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(6808..7028)
                     /label=URA3 promoter
     CDS             complement(7150..7941)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ggcttctcac gtcgttctca tcgtctgctt tctccccctt tccttttctt ccgtcgtttc
       61 tcccacagaa caattacggg agatgctgga tttctatgtg gaggctgacg ttttccgggt
      121 cagcgccgat cggacgatga ttgaactctt caacacggat tgagagagca ctgtacgcag
      181 tgcagagcgt ggtaggacat agaatcgcgc agggaaatag gttggaaggg gagcaattcg
      241 cagaaggaaa gctcagcgat cgagaaaaac atggagggtg aagataagct cgctctgaaa
      301 gtaaatctga caaatcaagc acagatgagg tcactgcgaa gttgacctga aagcgtgttt
      361 ttcaaatcgt cgcccaggcg aagtgttatt atttcttgca catcttgaag atagtctata
      421 cgcaggatga aaaggaggat caactttgcc atttctgcat acgtcataat tccacgatga
      481 ctagcaatgt ggaatagatc ttccttttga atttctctga atggttagca ctttcctttg
      541 tttttttgtc ctatgaaatt taaatggtaa ttgatacctc cactaaggag tcaaatcaat
      601 agctttgatc acaagcagaa gaaatgataa gagcgcaaag cttggaataa tctctttata
      661 tgtcccattg attaacatct atcctacaag ggcgtgaaca accacgatca gagtttagga
      721 gagcgatcaa ctccaagcac taaaagggcg tgaacaacca ctataagaat tgaagaggcg
      781 atcctccaag aacgctgccc gaccaggcat taattgccgc ataatctgac cgtcgtctgg
      841 atagcctggc agacgggaaa acttcatgga ataatgacgc cattacttga agtttcgcgg
      901 ggcttttttt cctggctcgg aattatactt tttacgccat aactgcgttt ccccgttctc
      961 caagggctcc gccgtcggca actttgtatg gggaagcgcg gtagattttg ttttcaagga
     1021 aggaccaagg gtgtgaggtg gcttttgaaa cactggtgcc atgtatggga ctattctgac
     1081 ggatctccgc cgcgcaactt aactcccgcc tcggattccg tgttaggcgt cgtaacgaac
     1141 attaggaaga catcggatct tagatgagaa attatgcatc atgcgacttg tgtgaatgtg
     1201 catcaatctt ttgcctgttg cattacgcat gctcacgcga cttcatgaat atgtcttccc
     1261 ttacatactc ctcctgatca cgacaacaca acatttctca catcactcca gggtggtcga
     1321 tggcggccgc aggatcagct gggcgcgccg ctggactgga agtgctgttt caaggaccag
     1381 ccggggatta caaagatgac gacgataagg ccggtgctca tcatcatcac catcaccatc
     1441 accaccatta atagcggcgc ttttgaccgg gaactcacgc atttaatgta ttcgaaacac
     1501 gctttttaag atctcaaatg aaacatgtat ccgtatgctc tccacatccg tcctcacgta
     1561 cggtgtgatg tttttttaat tttttatatg aaagtgtaag gcttctccat tttacttttt
     1621 gtgtttctgt cttcacagtc tttctctgcg agctgtgaga gggtcatgcc ctttatgggc
     1681 acgacggcag gattttggtt tttgaaagaa tttctgtcgg ctgtaagtct tcaaaaaaga
     1741 agaaagcggg gacggaaact tcttgcttgt gtgttataca ttacaaagtc ctctacaatt
     1801 ctacagtcct agttgttcac gcgacttcac gcagcagacc agatttatag ctttgcatga
     1861 tcgtctccga gcatttcaga atgctataat atggaatgat cttccgctgg cacctcatta
     1921 tgcgatacgc actccatata aacacaggta tccctgtccg tattcgttaa ctgttaattc
     1981 atgacacaac atcctgctgt atgattttgg cacaacgacg cggttggagg gtgagggaga
     2041 tggtggtgag acatcagaaa tggagagaat aacggtagag aaaaaaagga acacacgcgc
     2101 ccaggataga accaggacca cattccaagt ccctcaccag caacaagagg gacaccacgt
     2161 cggtgaagag gtgcgtgaga cagaggacac cagacaacgt gcacaaagcg tcggaaaggg
     2221 aggcgaccgc ttcgctggag gggggccgtc cacagcctgc gcccaaccac acacattcct
     2281 cccgcgaggc tcaagtgcca tgccgccgag tgccatgctc ctgcatccgg ggcaggtcga
     2341 cgagccagca tggtttggtc gccgacaacg cagatttgat ggagcttggg catttcttta
     2401 ccggacacgt cgcttcttac atttttacat tccaattcag aggtcacacc cgcgatcgag
     2461 aaagcagaga gtcggacccc cttcaagcag gaaaggcaag caacataccc atcgttaagt
     2521 ctgatgatgg ctggtgcaaa gccaaggcag ccagtcgata tgttaatgat cgggtacgtt
     2581 taaaggcgtg ggcaaggtta tttaacgctg gtagcatcgt agagttttta acgcaactga
     2641 agatttcatg tctgacttgg agatacggtc ggggcagcag ctatgggtaa aaagcctgaa
     2701 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     2761 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     2821 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     2881 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag
     2941 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     3001 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc
     3061 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     3121 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     3181 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     3241 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     3301 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     3361 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     3421 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
     3481 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     3541 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     3601 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
     3661 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata atgataacag
     3721 cgcttagagc tcgaatttcc ccgatcgttc aaacatttgg caataaagtt tcttaagatt
     3781 gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca
     3841 tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt
     3901 cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa
     3961 attatcgcgc gcggtgtcat ctatgttact agatcgggaa ttggttccgg aaccaattcg
     4021 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4081 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4141 ttaggctgag tttaaacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4201 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4261 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4321 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4381 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4441 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4501 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4561 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4621 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4681 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4741 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4801 tcctggcctt ttgctggcct tttgctcaca tcgttttcgg aaacgctttg aagttcctat
     4861 tccgaagttc ctattctcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag
     4921 cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa
     4981 atattgcgaa taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct
     5041 gtgctatatc cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac
     5101 tcgacctcta cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag
     5161 taagaactat tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt
     5221 agtatataga gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc
     5281 aacgctatca ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg
     5341 gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg
     5401 aagtggagtc aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa
     5461 ccttaacgga cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg
     5521 agaaaaaaag taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg
     5581 tagaacaaaa aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc
     5641 tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat
     5701 ttttgtttta caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc
     5761 atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt
     5821 tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt caggtggcac ttttcgggga
     5881 aatgtgcgcg gaacccctat ttggtgcggt atttcacacc gcatagggta ataactgata
     5941 taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt
     6001 tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg
     6061 ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat
     6121 gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc
     6181 cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc
     6241 acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc
     6301 aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa
     6361 catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac
     6421 aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta
     6481 tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc
     6541 gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat
     6601 ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc
     6661 ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt
     6721 ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc
     6781 acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct
     6841 gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat
     6901 atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga
     6961 atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt
     7021 gaattgaaaa gctgtggtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7081 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctcgt
     7141 ttaaactcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     7201 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     7261 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     7321 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     7381 ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc
     7441 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     7501 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     7561 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     7621 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     7681 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     7741 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     7801 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     7861 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     7921 caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagagcttga
     7981 tcccctgcgc catcagatcc ttggcggcga gaaagccatc cagtttactt tgcagggctt
     8041 cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg tccagtttaa
     8101 ac
//
