LOCUS       V004221                 6831 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004221
VERSION     V004221
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6831)
  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  JOURNAL   Unpublished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 6831)
  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 6831)
  AUTHORS   Lauersen KJ, Mussgnug JH, Kruse O.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAY-2016) Algae Biotechnology and Bioenergy,
            University of Bielefeld Center for Biotechnology, Universitaetsstr
            27, Bielefeld, NRW 33615, Germany
REFERENCE   4  (bases 1 to 6831)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6831)
  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            "Unpublished"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"; journalName: "Submitted
            (18-MAY-2016) Algae Biotechnology and Bioenergy, University of
            Bielefeld Center for Biotechnology, Universitaetsstr 27, Bielefeld,
            NRW 33615, Germany"
            SGRef: number: 4; type: "Journal Article"
            On May 18, 2016 this sequence version replaced KM061067.1.
FEATURES             Location/Qualifiers
     source          1..6831
                     /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             2269..2280
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             2293..2316
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
     CDS             3266..4261
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     promoter        complement(4531..4549)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4570..4586)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4594..4610)
                     /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(4618..4648)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4663..4684)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4972..5560)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5734..6591)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6592..6696)
                     /label="AmpR promoter"
     rep_origin      complement(join(6723..6831,1..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 agggcagggt
     1261 gagcaagggc gaggagctgt tcaccggcgt ggtgcccatc ctggtggagc tggacggcga
     1321 cgtgaacggc cacaagttca gcgtgcgcgg cgagggcgag ggcgacgcca ccaacggcaa
     1381 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctggt
     1441 gaccaccttc ggctacggcg tggcctgctt cagccgctac cccgaccaca tgaagcagca
     1501 cgacttcttc aagagcgcca tgcccgaggg ctacgtgcag gagcgcacca tcagcttcaa
     1561 ggacgacggc acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1621 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggt gagcttgcgg ggttgcgagc
     1681 aacactccag caacgaacag tgcccaagtc aggaatctgc agtcagcctg ggctttcggc
     1741 ggctttttct tgggcaaaca gcttgcactc atgccagcgc ggcttgtcca gcctcacttg
     1801 agctttccag ctgctaccag ccgggctata cgacagcgac agagccatag cgtggaatca
     1861 cttatttggg ttgccgaagt agcggtcgga gcgtgagttc ttggtcaagc cgccccttat
     1921 ccggttcctg tccgtgtctt tgtccctcgt tcacccttcg cggcaccctt catccccttg
     1981 cttgcaggta acatcctggg ccacaagctg gagtacaact tcaacagcca caacgtgtac
     2041 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacgtg
     2101 gaggacggca gcgtgcagct ggccgaccac taccagcaga acacccccat cggcgacggc
     2161 cccgtgctgc tgcccgacaa ccactacctg agccaccaga gcgccctgag caaggacccc
     2221 aacgagaagc gcgaccacat ggtgctgctg gagttcgtga ccgccgccat cgagggcagg
     2281 gatatcgaat tctggagcca cccgcagttc gagaagtaac cgctccgtgt aaatggaggc
     2341 gctcgttgat ctgagccttg ccccctgacg aacggcggtg gatggaagat actgctctca
     2401 agtgctgaag cggtagctta gctccccgtt tcgtgctgat cagtcttttt caacacgtaa
     2461 aaagcggagg agttttgcaa ttttgttggt tgtaacgatc ctccgttgat tttggcctct
     2521 ttctccatgg gcgggctggg cgtatttgaa gcgggatcct tcgaacgtac ggttaactcg
     2581 cgacctgcag gtacgtaatt taaatacgcg tgctgaggct tgacatgatt ggtgcgtatg
     2641 tttgtatgaa gctacaggac tgatttggcg ggctatgagg gcgggggaag ctctggaagg
     2701 gccgcgatgg ggcgcgcggc gtccagaagg cgccatacgg cccgctggcg gcacccatcc
     2761 ggtataaaag cccgcgaccc cgaacggtga cctccacttt cagcgacaaa cgagcactta
     2821 tacatacgcg actattctgc cgctatacat aaccactcag ctagcttaag atcccatccc
     2881 tagggcatgc cgggcgcgcc agaaggagcg cagccaaacc aggatgatgt ttgatggggt
     2941 atttgagcac ttgcaaccct tatccggaag ccccctggcc cacaaaggct aggcgccaat
     3001 gcaagcagtt cgcatgcagc ccctggagcg gtgccctcct gataaaccgg ccagggggcc
     3061 tatgttcttt acttttttac aagagaagtc actcaacatc ttaaaatggc caggtgagtc
     3121 gacgagcaag cccggcggat caggcagcgt gcttgcagat ttgacttgca acgcccgcat
     3181 tgtgtcgacg aaggcttttg gctcctctgt cgctgtctca agcagcatct aaccctgcgt
     3241 cgccgtttcc atttgcagga agcttatgac acaagaatcc ctgttacttc tcgaccgtat
     3301 tgattcggat gattcctacg cgagcctgcg gaacgaccag gagttctggg agccgctggc
     3361 ccgccgagcc ctggaggagc tcgggctgcc ggtgccgccg gtgctgcggg tgcccggcga
     3421 gagcaccaac cccgtactgg tcggcgagcc cggcccggtg atcaagctgt tcggcgagca
     3481 ctggtgcggt ccggagagcc tcgcgtcgga gtcggaggcc tacgcggtcc tggcggacgc
     3541 cccggtgccg gtgccccgcc tcctcggccg cggcgagctg cggcccggca ccggagcctg
     3601 gccgtggccc tacctggtga tgagccggat gaccggcacc acctggcggt ccgcgatgga
     3661 cggcacgacc gaccggaacg cgctgctcgc cctggcccgc gaactcggcc gggtgctcgg
     3721 ccggctgcac agggtgccgc tgaccgggaa caccgtgctc accccccatt ccgaggtctt
     3781 cccggaactg ctgcgggaac gccgcgcggc gaccgtcgag gaccaccgcg ggtggggcta
     3841 cctctcgccc cggctgctgg accgcctgga ggactggctg ccggacgtgg acacgctgct
     3901 ggccggccgc gaaccccggt tcgtccacgg cgacctgcac gggaccaaca tcttcgtgga
     3961 cctggccgcg accgaggtca ccgggatcgt cgacttcacc gacgtgtatg cgggagactc
     4021 ccgctacagc ctggtgcaac tgcatctcaa cgccttccgg ggcgaccgcg agatcctggc
     4081 cgcgctgctc gacggggcgc agtggaagcg gaccgaggac ttcgcccgcg aactgctcgc
     4141 cttcaccttc ctgcacgact tcgaggtgtt cgaggagacc ccgctggatc tctccggctt
     4201 caccgatccg gaggaactgg cgcagttcct ctgggggccg ccggacaccg cccccggcgc
     4261 ctgactcgag tgaccgctcc gtgtaaatgg aggcgctcgt tgatctgagc cttgccccct
     4321 gacgaacggc ggtggatgga agatactgct ctcaagtgct gaagcggtag cttagctccc
     4381 cgtttcgtgc tgatcagtct ttttcaacac gtaaaaagcg gaggagtttt gcaattttgt
     4441 tggttgtaac gatcctccgt tgattttggc ctctttctcc atgggcgggc tgggcgtatt
     4501 tgaagcggac ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     4561 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4621 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4681 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4741 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4801 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4861 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4921 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4981 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5041 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5101 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5161 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5221 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5281 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5341 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5401 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5461 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5521 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5581 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5641 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5701 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5761 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5821 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5881 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5941 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6001 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6061 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6121 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6181 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6241 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6301 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6361 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6421 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6481 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6541 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6601 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6661 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccaca
     6721 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     6781 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag a
//
