LOCUS       62056_18280        6014 bp DNA     circular SYN 17-APR-2021
DEFINITION  Cloning vector pOLK4, complete sequence.
ACCESSION   MW598458
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6014)
  AUTHORS   Thomy J, Sanchez F, Gut M, Cruz F, Alioto T, Piganeau G, Grimsley N,
            Yau S.
  TITLE     Combining Nanopore and Illumina Sequencing Permits Detailed Analysis
            of Insertion Mutations and Structural Variations Produced by 
            PEG-Mediated Transformation in Ostreococcus tauri
  JOURNAL   Cells 10 (3), 664 (2021)
  PUBMED    33802698
REFERENCE   2  (bases 1 to 6014)
  AUTHORS   Thomy J, Sanchez F, Gut M, Cruz F, Alioto T, Piganeau G, Grimsley N,
            Yau S.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-FEB-2021) UMR7232, CNRS, Avenue Pierre Fabre, Banyuls 
            sur mer 66650, France
REFERENCE   3  (bases 1 to 6014)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cells"; 
            date: "2021"; volume: "10"; issue: "3"; pages: "664"
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..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      725..1313
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1601..1622
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1637..1667
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1675..1691
                     /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     1699..1715
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2133..3782
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             4186..4992
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5113..5129)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5603..5707
                     /label=AmpR promoter
     CDS             join(5708..6014,1..551)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
       61 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      121 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      181 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      241 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      301 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      361 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      421 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      481 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      541 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      601 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      721 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      781 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      841 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      901 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      961 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1021 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1081 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1141 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1201 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1261 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1321 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1381 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1441 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     1501 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     1561 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     1621 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     1681 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctttgatcat
     1741 agctgagtaa cgcgacgacg tgtagtgagt gtaatgatgg atcgtgtata gcgtagtatt
     1801 aatgggctca cgcagccttc ggtcgacgcg ctatcgctcg aaaacgcgtg tgaatgcaca
     1861 ctctcctccg tcacctagtt aagatctcgt gcgcgtcgtc catcatactc tggacggaaa
     1921 acgcccatcg acgtcacaca cgcaccgacc atcatactct ggacggaaaa cgcccatcga
     1981 cgtcacacac ggcccgtggt cggggccccc ggtgccgtcc gtgtgccaat gatggacgat
     2041 atgtgtctcc aatacgagac ccccgatccc cggacccgcg gtcaaccacc gcgcgtcgac
     2101 gcgacggcga aaccgcacac gcgcgcgcgc acatggagga cgcgaagaac atcaagaagg
     2161 ggccggcgcc gttctacccg ctggaggacg ggacggcggg ggagcagctg cacaaggcga
     2221 tgaagcgcta cgcgctggtg ccggggacga tcgcgttcac ggacgcgcac atcgaggtgg
     2281 acatcacgta cgcggaatac ttcgagatgt ccgtgcgcct ggcggaggcg atgaagcgct
     2341 acgggctgaa cacgaaccac cgcatcgtgg tgtgctccga gaactccctg cagttcttca
     2401 tgccggtgct gggggcgctg ttcatcgggg tggcggtggc gccggcgaac gacatctaca
     2461 acgagcgcga gctgctgaac tcgatgggga tctcccagcc gacggtggtg ttcgtgtcca
     2521 agaaggggct gcagaagatc ctgaacgtgc agaagaagct gccgatcatc cagaagatca
     2581 tcatcatgga ctccaagacg gactaccagg ggttccagtc catgtacacg ttcgtgacgt
     2641 cccacctgcc gccggggttc aacgagtacg acttcgtgcc ggagtccttc gaccgcgaca
     2701 agacgatcgc gctgatcatg aactcctccg ggtccacggg gctgccgaag ggggtggcgc
     2761 tgccgcaccg cacggcgtgc gtgcgcttct cccacgcgcg cgacccgatc ttcgggaacc
     2821 agatcatccc ggacacggcg atcctgtccg tggtgccgtt ccaccacggg ttcgggatgt
     2881 tcacgacgct gggctacctg atctgcgggt tccgcgtggt gctgatgtac cgcttcgagg
     2941 aggagctgtt cctgcgctcc ctgcaggact acaagatcca gtccgcgctg ctggtgccga
     3001 cgctgttctc cttcttcgcg aagtccacgc tgatcgacaa gtacgacctg tccaacctgc
     3061 acgagatcgc gtccgggggg gcgccgctgt ccaaggaggt gggggaggcg gtggcgaagc
     3121 gcttccacct gccgggcatc cgccaggggt acgggctgac ggagacgacg tccgcgatcc
     3181 tgatcacgcc ggagggggac gacaagccgg gggcggtggg gaaggtggtg ccgttcttcg
     3241 aggcgaaggt ggtggacctg gacacgggga agacgctggg ggtgaaccag cgcggggagc
     3301 tgtgcgtgcg cgggccgatg atcatgtccg ggtacgtgaa caacccggag gcgacgaacg
     3361 cgctgatcga caaggacggg tggctgcact ccggggacat cgcgtactgg gacgaggacg
     3421 agcacttctt catcgtggac cgcctgaagt ccctgatcaa gtacaagggc taccaggtgg
     3481 cgccggcgga gctggagtcc atcctgctgc agcacccgaa catcttcgac gcgggggtgg
     3541 cggggctgcc ggacgacgac gcgggggagc tgccggcggc ggtggtggtg ctggagcacg
     3601 ggaagacgat gacggagaag gagatcgtgg actacgtggc gtcccaggtg acgacggcga
     3661 agaagctgcg cgggggggtg gtgttcgtgg acgaggtgcc gaagggcctg acggggaagc
     3721 tggacgcgcg caagatccgc gagatcctga tcaaggcgaa gaaggggggg aagatcgcgg
     3781 tgtgatcgat cgagttttac gatcacgcta tcgcaccgcg gtcgacacag tagatggaaa
     3841 taaacgcaaa tgtaccactc acactcggat tcgaacgatt tcaaactgtt cagcatggat
     3901 gaaaaaaacg accaaacgga caaacgagcc gtattgaata acgtatacat ctatgaattc
     3961 aatgaagaaa aggggaacgg ttgtgatcgt cgcaacgtcc acgaaaccgt actcctcgac
     4021 gcgacgccgc agcgacatcg actcgctcgc cgacgcatct ctcgagaagt aattgttctc
     4081 gtatttttac ggttaccaac cgtcccgcag acgctcgcgt gcggtacacg aagagtatct
     4141 ttaagaaact cgaccaacgt ttgaatcttt tttgattcga caaagatggg caaggagaag
     4201 acgcacgtga gccgcccgcg cctcaacagc aacatggacg cggacctgta cgggtacaag
     4261 tgggcgcgcg acaacgtggg gcagagcggg gcgacgatct acaggctgta cgggaagccg
     4321 gacgcgccgg agctgttcct gaagcacggg aagggcagcg tggcgaacga cgtgacggac
     4381 gagatggtgc gcctgaactg gctgacggag ttcatgccgc tgccgacgat caagcacttc
     4441 atccgcacgc cggacgacgc gtggctgctg acgacggcga ttccggggaa gacggcgttc
     4501 caggtgctgg aggagtaccc ggacagcggc gagaacatcg tggacgcgct ggcggtgttc
     4561 ctgcgccgcc tgcacagcat cccggtgtgc aactgcccgt tcaacagcga ccgcgtgttc
     4621 cgcctggcgc aggcgcagag ccgcatgaac aacgggctgg tggacgcgag cgacttcgac
     4681 gacgagcgca acgggtggcc ggtggagcag gtgtggaagg agatgcacaa gctgctgccg
     4741 ttcagcccgg acagcgtggt gacgcacggg gacttcagcc tggacaacct gatcttcgac
     4801 gaggggaagc tgatcgggtg catcgacgtg gggcgcgtgg ggatcgcgga ccgctaccag
     4861 gacctggcga tcctgtggaa ctgcctgggg gagttctcgc cgtcgttgca gaagcgcctg
     4921 ttccagaagt acgggatcga caacccggac atgaacaagc tccagttcca cctgatgctg
     4981 gacgagttct tctagcttgt cgagttttac gatcacgcta tcgcaccgcg gtcgacacag
     5041 tagatggaaa taaacgcaaa tgtaccactc acactcatct agatgcattc gcgaggtacc
     5101 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5161 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5221 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     5281 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     5341 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5401 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5461 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5521 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5581 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5641 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5701 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5761 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5821 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5881 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5941 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6001 tgacttggtt gagt
//
