LOCUS       V004219                 6657 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004219
VERSION     V004219
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6657)
  AUTHORS   Lauersen KJ, Mussgnug JH, Kruse O.
  TITLE     Efficient targeted expression of nuclear transgenes in Chlamydomonas
            reinhardtii with a novel, modular vector toolkit
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6657)
  AUTHORS   Lauersen KJ, Mussgnug JH, Kruse O.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2014) Algae Biotechnology and Bioenergy,
            University of Bielefeld Center for Biotechnology, Universitaetsstr
            27, Bielefeld, NRW 33615, Germany
REFERENCE   3  (bases 1 to 6657)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6657)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (25-JUN-2014) Algae Biotechnology and Bioenergy, University of
            Bielefeld Center for Biotechnology, Universitaetsstr 27, Bielefeld,
            NRW 33615, Germany"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     489..505
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        515..533
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1247..1258
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             2095..2106
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             2119..2142
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
     CDS             3092..4087
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     promoter        complement(4357..4375)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4396..4412)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4420..4436)
                     /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(4444..4474)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4489..4510)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4798..5386)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5560..6417)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6418..6522)
                     /label="AmpR promoter"
     rep_origin      complement(join(6549..6657,1..347))
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
       61 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
      121 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
      181 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
      241 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
      301 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
      361 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
      421 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
      481 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
      541 ggagctccac cgcggtggcg gccgctctag aactagtgct gaggcttgac atgattggtg
      601 cgtatgtttg tatgaagcta caggactgat ttggcgggct atgagggcgg gggaagctct
      661 ggaagggccg cgatggggcg cgcggcgtcc agaaggcgcc atacggcccg ctggcggcac
      721 ccatccggta taaaagcccg cgaccccgaa cggtgacctc cactttcagc gacaaacgag
      781 cacttataca tacgcgacta ttctgccgct atacataacc actcagctag cttaagatcc
      841 catcaccggt gcatgccggg cgcgccagaa ggagcgcagc caaaccagga tgatgtttga
      901 tggggtattt gagcacttgc aacccttatc cggaagcccc ctggcccaca aaggctaggc
      961 gccaatgcaa gcagttcgca tgcagcccct ggagcggtgc cctcctgata aaccggccag
     1021 ggggcctatg ttctttactt ttttacaaga gaagtcactc aacatcttaa aatggccagg
     1081 tgagtcgacg agcaagcccg gcggatcagg cagcgtgctt gcagatttga cttgcaacgc
     1141 ccgcattgtg tcgacgaagg cttttggctc ctctgtcgct gtctcaagca gcatctaacc
     1201 ctgcgtcgcc gtttccattt gcaggatgca tatgagatct gacgtcatcg agggcaggaa
     1261 gccgacggag aacaacgagg acttcaacat tgtcgccgtg gcgtccaact ttgccaccac
     1321 cgatctggac gccgaccgcg gcaagctgcc gggcaagaag ctgcccctgg aggtcctcaa
     1381 ggaaatggag gcgaacgccc ggaaggccgg ctgcacccgc ggctgcctga tctgcctgtc
     1441 ccacatcaag tgcacgccga agatgaagaa gttcattccc ggccggtgag cttgcggggt
     1501 tgcgagcaac actccagcaa cgaacagtgc ccaagtcagg aatctgcagt cagcctgggc
     1561 tttcggcggc tttttcttgg gcaaacagct tgcactcatg ccagcgcggc ttgtccagcc
     1621 tcacttgagc tttccagctg ctaccagccg ggctatacga cagcgacaga gccatagcgt
     1681 ggaatcactt atttgggttg ccgaagtagc ggtcggagcg tgagttcttg gtcaagccgc
     1741 cccttatccg gttcctgtcc gtgtctttgt ccctcgttca cccttcgcgg cacccttcat
     1801 ccccttgctt gcaggtgcca cacctacgag ggcgacaagg agtccgcgca gggcggcatt
     1861 ggcgaggcga tcgtggacat ccccgagatt ccgggcttca aggacctgga gccgatggag
     1921 cagttcatcg cccaggtgga cctgtgcgtg gactgcacga ccggctgtct gaagggcctg
     1981 gccaatgtgc agtgttccga cctgctcaag aagtggctgc cccagcgctg tgccaccttc
     2041 gccagcaaga tccagggcca ggtggacaag attaagggcg ctggcggcga tacgatcgag
     2101 ggcagggata tcgaattctg gagccacccg cagttcgaga agtaaccgct ccgtgtaaat
     2161 ggaggcgctc gttgatctga gccttgcccc ctgacgaacg gcggtggatg gaagatactg
     2221 ctctcaagtg ctgaagcggt agcttagctc cccgtttcgt gctgatcagt ctttttcaac
     2281 acgtaaaaag cggaggagtt ttgcaatttt gttggttgta acgatcctcc gttgattttg
     2341 gcctctttct ccatgggcgg gctgggcgta tttgaagcgg gatccttcga acgtacggtt
     2401 aactcgcgac ctgcaggtac gtaatttaaa tacgcgtgct gaggcttgac atgattggtg
     2461 cgtatgtttg tatgaagcta caggactgat ttggcgggct atgagggcgg gggaagctct
     2521 ggaagggccg cgatggggcg cgcggcgtcc agaaggcgcc atacggcccg ctggcggcac
     2581 ccatccggta taaaagcccg cgaccccgaa cggtgacctc cactttcagc gacaaacgag
     2641 cacttataca tacgcgacta ttctgccgct atacataacc actcagctag cttaagatcc
     2701 catccctagg gcatgccggg cgcgccagaa ggagcgcagc caaaccagga tgatgtttga
     2761 tggggtattt gagcacttgc aacccttatc cggaagcccc ctggcccaca aaggctaggc
     2821 gccaatgcaa gcagttcgca tgcagcccct ggagcggtgc cctcctgata aaccggccag
     2881 ggggcctatg ttctttactt ttttacaaga gaagtcactc aacatcttaa aatggccagg
     2941 tgagtcgacg agcaagcccg gcggatcagg cagcgtgctt gcagatttga cttgcaacgc
     3001 ccgcattgtg tcgacgaagg cttttggctc ctctgtcgct gtctcaagca gcatctaacc
     3061 ctgcgtcgcc gtttccattt gcaggaagct tatgacacaa gaatccctgt tacttctcga
     3121 ccgtattgat tcggatgatt cctacgcgag cctgcggaac gaccaggagt tctgggagcc
     3181 gctggcccgc cgagccctgg aggagctcgg gctgccggtg ccgccggtgc tgcgggtgcc
     3241 cggcgagagc accaaccccg tactggtcgg cgagcccggc ccggtgatca agctgttcgg
     3301 cgagcactgg tgcggtccgg agagcctcgc gtcggagtcg gaggcctacg cggtcctggc
     3361 ggacgccccg gtgccggtgc cccgcctcct cggccgcggc gagctgcggc ccggcaccgg
     3421 agcctggccg tggccctacc tggtgatgag ccggatgacc ggcaccacct ggcggtccgc
     3481 gatggacggc acgaccgacc ggaacgcgct gctcgccctg gcccgcgaac tcggccgggt
     3541 gctcggccgg ctgcacaggg tgccgctgac cgggaacacc gtgctcaccc cccattccga
     3601 ggtcttcccg gaactgctgc gggaacgccg cgcggcgacc gtcgaggacc accgcgggtg
     3661 gggctacctc tcgccccggc tgctggaccg cctggaggac tggctgccgg acgtggacac
     3721 gctgctggcc ggccgcgaac cccggttcgt ccacggcgac ctgcacggga ccaacatctt
     3781 cgtggacctg gccgcgaccg aggtcaccgg gatcgtcgac ttcaccgacg tgtatgcggg
     3841 agactcccgc tacagcctgg tgcaactgca tctcaacgcc ttccggggcg accgcgagat
     3901 cctggccgcg ctgctcgacg gggcgcagtg gaagcggacc gaggacttcg cccgcgaact
     3961 gctcgccttc accttcctgc acgacttcga ggtgttcgag gagaccccgc tggatctctc
     4021 cggcttcacc gatccggagg aactggcgca gttcctctgg gggccgccgg acaccgcccc
     4081 cggcgcctga ctcgagtgac cgctccgtgt aaatggaggc gctcgttgat ctgagccttg
     4141 ccccctgacg aacggcggtg gatggaagat actgctctca agtgctgaag cggtagctta
     4201 gctccccgtt tcgtgctgat cagtcttttt caacacgtaa aaagcggagg agttttgcaa
     4261 ttttgttggt tgtaacgatc ctccgttgat tttggcctct ttctccatgg gcgggctggg
     4321 cgtatttgaa gcggacccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg
     4381 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4441 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4501 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4561 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4621 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4681 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4981 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5221 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5401 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5461 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5521 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5581 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5641 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5701 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5761 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5821 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5881 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5941 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6001 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6061 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6121 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6181 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6241 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6301 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6361 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6421 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6481 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6541 gccacactaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     6601 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaaga
//
