LOCUS       62056_18275        6125 bp DNA     circular SYN 17-APR-2021
DEFINITION  Cloning vector pOLK1, complete sequence.
ACCESSION   MW598457
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6125)
  AUTHORS   Sanchez F, Geffroy S, Norest M, Yau S, Moreau H, Grimsley N.
  TITLE     Simplified Transformation of Ostreococcus tauri Using Polyethylene 
            Glycol
  JOURNAL   Genes (Basel) 10 (5), E399 (2019)
  PUBMED    31130696
REFERENCE   2  (bases 1 to 6125)
  AUTHORS   Sanchez F, Geffroy S, Norest M, Yau S, Moreau H, Grimsley N.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-FEB-2021) UMR7232, CNRS, Avenue Pierre Fabre, Banyuls 
            sur mer 66650, France
REFERENCE   3  (bases 1 to 6125)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genes 
            (Basel)"; date: "2019"; volume: "10"; issue: "5"; pages: "E399"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-FEB-2021) UMR7232, CNRS, Avenue Pierre Fabre, Banyuls sur mer 
            66650, France"
FEATURES             Location/Qualifiers
     source          1..6125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2235..2240
                     /label=restriction site HindIII
                     /note="restriction site HindIII"
     regulatory      2247..2388
                     /label=strong stop terminator
                     /note="strong stop terminator"
                     /regulatory_class="terminator"
     misc_feature    2389..2394
                     /label=restriction site BglII
                     /note="restriction site BglII"
     regulatory      2395..2708
                     /label=alpha tubulin promoter
                     /note="alpha tubulin promoter"
                     /regulatory_class="promoter"
     misc_feature    2587..2592
                     /label=restriction site XhoI
                     /note="restriction site XhoI"
     misc_feature    2617..2623
                     /label=restriction site BstEII
                     /note="restriction site BstEII"
     CDS             2712..4361
                     /label=luciferase
                     /note="firefly luciferase"
     misc_feature    4364..4369
                     /label=restriction site ClaI
                     /note="restriction site ClaI"
     regulatory      4370..4445
                     /label=alpha tubulin terminator
                     /note="alpha tubulin terminator"
                     /regulatory_class="terminator"
     misc_feature    4446..4451
                     /label=restriction site BamHI
                     /note="restriction site BamHI"
     misc_feature    4450..4455
                     /label=restriction site SmaI/XmaI
                     /note="restriction site SmaI/XmaI"
     misc_feature    4470..4475
                     /label=restriction site StuI
                     /note="restriction site StuI"
     misc_feature    4476..4481
                     /label=restriction site SphI
                     /note="restriction site SphI"
     regulatory      4485..4799
                     /label=alpha tubulin promoter
                     /note="alpha tubulin promoter"
                     /regulatory_class="promoter"
     misc_feature    4675..4680
                     /label=restriction site XhoI
                     /note="restriction site XhoI"
     misc_feature    4705..4711
                     /label=restriction site BstEII
                     /note="restriction site BstEII"
     CDS             4800..5606
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      5615..5690
                     /label=alpha tubulin terminator
                     /note="alpha tubulin terminator"
                     /regulatory_class="terminator"
     misc_feature    5696..5701
                     /label=restriction site XbaI
                     /note="restriction site XbaI"
     misc_feature    5707..5712
                     /label=restriction site NruI
                     /note="restriction site NruI"
     misc_feature    5713..5718
                     /label=restriction site KpnI
                     /note="restriction site KpnI"
     misc_feature    5719..5724
                     /label=multiple cloning site: SacI/SstI
                     /note="multiple cloning site: SacI/SstI"
     primer_bind     complement(5731..5747)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5871..5881
                     /label=restriction site BglI
                     /note="restriction site BglI"
     misc_feature    5938..5943
                     /label=restriction site NdeI
                     /note="restriction site NdeI"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt tgatcatagc tgagtaacgc gacgacgtgt agtgagtgta
     2281 atgatggatc gtgtatagcg tagtattaat gggctcacgc agccttcggt cgacgcgcta
     2341 tcgctcgaaa acgcgtgtga atgcacactc tcctccgtca cctagttaag atctagtcga
     2401 acgatttcaa actgttcagc atggatgaaa aaaacgacca aacggacaaa cgagccgtat
     2461 tgaataacgt atacatctat gaattcaatg aagaaaaggg gaacggttgt gatcgtcgca
     2521 acgtccacga aaccgtactc ctcgacgcga cgccgcagcg acatcgactc gctcgccgac
     2581 gcatctctcg agaagtaatt gttctcgtat ttttacggtt accaaccgtc ccgcagacgc
     2641 tcgcgtgcgg tacacgaaga gtatctttaa gaaactcgac caacgtttga atcttttttg
     2701 attcgacaac catggaggac gcgaagaaca tcaagaaggg gccggcgccg ttctacccgc
     2761 tggaggacgg gacggcgggg gagcagctgc acaaggcgat gaagcgctac gcgctggtgc
     2821 cggggacgat cgcgttcacg gacgcgcaca tcgaggtgga catcacgtac gcggaatact
     2881 tcgagatgtc cgtgcgcctg gcggaggcga tgaagcgcta cgggctgaac acgaaccacc
     2941 gcatcgtggt gtgctccgag aactccctgc agttcttcat gccggtgctg ggggcgctgt
     3001 tcatcggggt ggcggtggcg ccggcgaacg acatctacaa cgagcgcgag ctgctgaact
     3061 cgatggggat ctcccagccg acggtggtgt tcgtgtccaa gaaggggctg cagaagatcc
     3121 tgaacgtgca gaagaagctg ccgatcatcc agaagatcat catcatggac tccaagacgg
     3181 actaccaggg gttccagtcc atgtacacgt tcgtgacgtc ccacctgccg ccggggttca
     3241 acgagtacga cttcgtgccg gagtccttcg accgcgacaa gacgatcgcg ctgatcatga
     3301 actcctccgg gtccacgggg ctgccgaagg gggtggcgct gccgcaccgc acggcgtgcg
     3361 tgcgcttctc ccacgcgcgc gacccgatct tcgggaacca gatcatcccg gacacggcga
     3421 tcctgtccgt ggtgccgttc caccacgggt tcgggatgtt cacgacgctg ggctacctga
     3481 tctgcgggtt ccgcgtggtg ctgatgtacc gcttcgagga ggagctgttc ctgcgctccc
     3541 tgcaggacta caagatccag tccgcgctgc tggtgccgac gctgttctcc ttcttcgcga
     3601 agtccacgct gatcgacaag tacgacctgt ccaacctgca cgagatcgcg tccggggggg
     3661 cgccgctgtc caaggaggtg ggggaggcgg tggcgaagcg cttccacctg ccgggcatcc
     3721 gccaggggta cgggctgacg gagacgacgt ccgcgatcct gatcacgccg gagggggacg
     3781 acaagccggg ggcggtgggg aaggtggtgc cgttcttcga ggcgaaggtg gtggacctgg
     3841 acacggggaa gacgctgggg gtgaaccagc gcggggagct gtgcgtgcgc gggccgatga
     3901 tcatgtccgg gtacgtgaac aacccggagg cgacgaacgc gctgatcgac aaggacgggt
     3961 ggctgcactc cggggacatc gcgtactggg acgaggacga gcacttcttc atcgtggacc
     4021 gcctgaagtc cctgatcaag tacaagggct accaggtggc gccggcggag ctggagtcca
     4081 tcctgctgca gcacccgaac atcttcgacg cgggggtggc ggggctgccg gacgacgacg
     4141 cgggggagct gccggcggcg gtggtggtgc tggagcacgg gaagacgatg acggagaagg
     4201 agatcgtgga ctacgtggcg tcccaggtga cgacggcgaa gaagctgcgc gggggggtgg
     4261 tgttcgtgga cgaggtgccg aagggcctga cggggaagct ggacgcgcgc aagatccgcg
     4321 agatcctgat caaggcgaag aaggggggga agatcgcggt gtgatcgatc gagttttacg
     4381 atcacgctat cgcaccgcgg tcgacacagt agatggaaat aaacgcaaat gtaccactca
     4441 cactcggatc ccgggcccgt cgactgcaga ggcctgcatg caagtcgaac gatttcaaac
     4501 tgttcagcat ggatgaaaaa aacgaccaaa cggacaaacg agccgtattg aataacgtat
     4561 acatctatga attcaatgaa gaaaagggga acggttgtga tcgtcgcaac gtccacgaaa
     4621 ccgtactcct cgacgcgacg ccgcagcgac atcgactcgc tcgccgacgc atctctcgag
     4681 aagtaattgt tctcgtattt ttacggttac caaccgtccc gcagacgctc gcgtgcggta
     4741 cacgaagagt atctttaaga aactcgacca acgtttgaat cttttttgat tcgacaaaga
     4801 tgggcaagga gaagacgcac gtgagccgcc cgcgcctcaa cagcaacatg gacgcggacc
     4861 tgtacgggta caagtgggcg cgcgacaacg tggggcagag cggggcgacg atctacaggc
     4921 tgtacgggaa gccggacgcg ccggagctgt tcctgaagca cgggaagggc agcgtggcga
     4981 acgacgtgac ggacgagatg gtgcgcctga actggctgac ggagttcatg ccgctgccga
     5041 cgatcaagca cttcatccgc acgccggacg acgcgtggct gctgacgacg gcgattccgg
     5101 ggaagacggc gttccaggtg ctggaggagt acccggacag cggcgagaac atcgtggacg
     5161 cgctggcggt gttcctgcgc cgcctgcaca gcatcccggt gtgcaactgc ccgttcaaca
     5221 gcgaccgcgt gttccgcctg gcgcaggcgc agagccgcat gaacaacggg ctggtggacg
     5281 cgagcgactt cgacgacgag cgcaacgggt ggccggtgga gcaggtgtgg aaggagatgc
     5341 acaagctgct gccgttcagc ccggacagcg tggtgacgca cggggacttc agcctggaca
     5401 acctgatctt cgacgagggg aagctgatcg ggtgcatcga cgtggggcgc gtggggatcg
     5461 cggaccgcta ccaggacctg gcgatcctgt ggaactgcct gggggagttc tcgccgtcgt
     5521 tgcagaagcg cctgttccag aagtacggga tcgacaaccc ggacatgaac aagctccagt
     5581 tccacctgat gctggacgag ttcttctagc ttgtcgagtt ttacgatcac gctatcgcac
     5641 cgcggtcgac acagtagatg gaaataaacg caaatgtacc actcacactc gatcatctag
     5701 atgcattcgc gaggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     5761 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5821 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5881 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     5941 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     6001 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     6061 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     6121 cgcga
//
