LOCUS       40924_27608        7888 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLCP.CFH, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7888)
  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            Nannochloropsis gaditana (unpublished)
  JOURNAL   Unpublished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 7888)
  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 7888)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7888)
  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            "Unpublished"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..7888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..1105
                     /label=LCP promoter
                     /note="LCP promoter"
                     /regulatory_class="promoter"
     misc_feature    1109..1116
                     /label=NotI restriction site
                     /note="NotI restriction site"
     misc_feature    1128..1135
                     /label=AscI restriction site
                     /note="AscI restriction site"
     CDS             1142..1165
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             1172..1195
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1205..1231
                     /label=9xHis
                     /note="9xHis affinity tag"
     regulatory      1238..1745
                     /label=ATPa terminator
                     /note="ATPa terminator"
                     /regulatory_class="terminator"
     regulatory      1776..2468
                     /label=UEP promoter
                     /note="UEP promoter"
                     /regulatory_class="promoter"
     CDS             2475..3494
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     terminator      3529..3781
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3815..3831)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3839..3855)
                     /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(3863..3893)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3908..3929)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      3970..4558
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4664..5646
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5793..6593)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(6594..6814)
                     /label=URA3 promoter
     CDS             complement(6936..7727)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 cgcattcagg tctagtaagc cgaagctaga ggcactcggc tcagtggcca cttgccaggc
       61 gcagttaggc catttgacgt cagttttgcc tcgatctgat ggcattggag ccaaaaaggg
      121 gtatttttta tcgaagtgct gatactgctg gctgggatga ggtactaaaa cggtagcacc
      181 agtcatgctg ggctcgccta caggaggtat agcaaagttt gtggcctcga gtacggcgtc
      241 gctggctgag cttaaaaaaa aaggtggaat atggagaatg aaggccaagt aggatagtat
      301 cgatttcccc atagttctgg ccttgtgtac agtacaaaga atgatgaaag ttagtttggt
      361 gagctgtcgc ttgtcgtggt caatgggccg tgggaacaac ctcccctgcc cttaagggag
      421 acaaaatcgt ggtacgtgct tgctcgataa agcccagatt cgtcctacgc atttatacaa
      481 aaatgccacc gctgttgtgc aacgcaacta cgagttaaat ccactatcgt gcgctggcgg
      541 aacaacgagc acggccgcaa caacatcgtg tgcagggccc tgctacctgg tattgttttg
      601 ttttctgtag ggaagacgtc gcttctttcc gtgttgctga ctcggcgacg agcatcagga
      661 aagcccaaga gacccaaccc ttcttcgtgc aggggaatgt cgtctgtagt atttcgttat
      721 tttcgtgaaa acgatatttg tgcggggggt cgagctggtg ggttgaggac tacatgcttg
      781 gcaccggagg ctggttttct ttcttccccc ttatgaattc gcctcgcgaa ccagaagcgg
      841 cgaattgctc atcgcggaga acatgacatt taaatcagaa tgtgccttgc atccttcacc
      901 acgtatcgct atatcttgca tctgaagtgc ttctacgcag cggtcatgat gaaattcttg
      961 agacattttg gtgtgttttg agtcctgtct tgaagcccgc acggagtggc tcataaggcg
     1021 aggaacaaat gtgatgccac tcccacctcg aaccattttt tcctatgtcc accctacgca
     1081 gccacacatt tgaacagaag caataatggc ggccgcagga tcagctgggc gcgccgctgg
     1141 actggaagtg ctgtttcaag gaccagccgg ggattacaaa gatgacgacg ataaggccgg
     1201 tgctcatcat catcaccatc accatcacca ccattaatag cggcgctttt gaccgggaac
     1261 tcacgcattt aatgtattcg aaacacgctt tttaagatct caaatgaaac atgtatccgt
     1321 atgctctcca catccgtcct cacgtacggt gtgatgtttt tttaattttt tatatgaaag
     1381 tgtaaggctt ctccatttta ctttttgtgt ttctgtcttc acagtctttc tctgcgagct
     1441 gtgagagggt catgcccttt atgggcacga cggcaggatt ttggtttttg aaagaatttc
     1501 tgtcggctgt aagtcttcaa aaaagaagaa agcggggacg gaaacttctt gcttgtgtgt
     1561 tatacattac aaagtcctct acaattctac agtcctagtt gttcacgcga cttcacgcag
     1621 cagaccagat ttatagcttt gcatgatcgt ctccgagcat ttcagaatgc tataatatgg
     1681 aatgatcttc cgctggcacc tcattatgcg atacgcactc catataaaca caggtatccc
     1741 tgtccgtatt cgttaactgt taattcatga cacaacatcc tgctgtatga ttttggcaca
     1801 acgacgcggt tggagggtga gggagatggt ggtgagacat cagaaatgga gagaataacg
     1861 gtagagaaaa aaaggaacac acgcgcccag gatagaacca ggaccacatt ccaagtccct
     1921 caccagcaac aagagggaca ccacgtcggt gaagaggtgc gtgagacaga ggacaccaga
     1981 caacgtgcac aaagcgtcgg aaagggaggc gaccgcttcg ctggaggggg gccgtccaca
     2041 gcctgcgccc aaccacacac attcctcccg cgaggctcaa gtgccatgcc gccgagtgcc
     2101 atgctcctgc atccggggca ggtcgacgag ccagcatggt ttggtcgccg acaacgcaga
     2161 tttgatggag cttgggcatt tctttaccgg acacgtcgct tcttacattt ttacattcca
     2221 attcagaggt cacacccgcg atcgagaaag cagagagtcg gacccccttc aagcaggaaa
     2281 ggcaagcaac atacccatcg ttaagtctga tgatggctgg tgcaaagcca aggcagccag
     2341 tcgatatgtt aatgatcggg tacgtttaaa ggcgtgggca aggttattta acgctggtag
     2401 catcgtagag tttttaacgc aactgaagat ttcatgtctg acttggagat acggtcgggg
     2461 cagcagctat gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     2521 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     2581 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     2641 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     2701 tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     2761 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     2821 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     2881 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     2941 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     3001 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     3061 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     3121 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     3181 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     3241 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     3301 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     3361 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     3421 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     3481 cgagggcaaa ggaataatga taacagcgct tagagctcga atttccccga tcgttcaaac
     3541 atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata
     3601 taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt
     3661 atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac
     3721 aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat
     3781 cgggaattgg ttccggaacc aattcgtaat catggtcata gctgtttcct gtgtgaaatt
     3841 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     3901 gtgcctaatg agtgagctaa ctcacattag gctgagttta aaccccgtag aaaagatcaa
     3961 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4021 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4081 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4141 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4201 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4261 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4321 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     4381 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4441 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4501 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4561 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatcgt
     4621 tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac
     4681 ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac
     4741 gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa acattgctca
     4801 aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac ccatccacct
     4861 ttcgctcctt gaacttgcat ctaaactcga cctctacatt ttttatgttt atctctagta
     4921 ttactcttta gacaaaaaaa ttgtagtaag aactattcat agagtgaatc gaaaacaata
     4981 cgaaaatgta aacatttcct atacgtagta tatagagaca aaatagaaga aaccgttcat
     5041 aattttctga ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa agtatgcgca
     5101 atccacatcg gtatagaata taatcgggga tgcctttatc ttgaaaaaat gcacccgcag
     5161 cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg gaagagaaaa
     5221 tagacaccaa agtagccttc ttctaacctt aacggaccta cagtgcaaaa agttatcaag
     5281 agactgcatt atagagcgca caaaggagaa aaaaagtaat ctaagatgct ttgttagaaa
     5341 aatagcgctc tcgggatgca tttttgtaga acaaaaaaga agtatagatt ctttgttggt
     5401 aaaatagcgc tctcgcgttg catttctgtt ctgtaaaaat gcagctcaga ttctttgttt
     5461 gaaaaattag cgctctcgcg ttgcattttt gttttacaaa aatgaagcac agattcttcg
     5521 ttggtaaaat agcgctttcg cgttgcattt ctgttctgta aaaatgcagc tcagattctt
     5581 tgtttgaaaa attagcgctc tcgcgttgca tttttgttct acaaaatgaa gcacagatgc
     5641 ttcgttcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgg tgcggtattt
     5701 cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     5761 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     5821 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     5881 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     5941 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     6001 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     6061 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     6121 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     6181 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     6241 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     6301 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     6361 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     6421 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     6481 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     6541 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     6601 cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     6661 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     6721 tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg aaatcaaaaa
     6781 aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg tggtatggtg cactctcagt
     6841 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6901 gcgccctgac gggcttgtct gctcgtttaa actcagaaga actcgtcaag aaggcgatag
     6961 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     7021 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     7081 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     7141 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc
     7201 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     7261 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     7321 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     7381 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     7441 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     7501 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg
     7561 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     7621 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     7681 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     7741 catcctgtct cttgatcaga gcttgatccc ctgcgccatc agatccttgg cggcgagaaa
     7801 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     7861 tccggttcgc ttgctgtcca gtttaaac
//
