LOCUS       V004217                 6864 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004217
VERSION     V004217
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6864)
  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  JOURNAL   Unpublished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 6864)
  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 6864)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6864)
  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            "Unpublished"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..6864
                     /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             2302..2313
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             2326..2349
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
     CDS             3299..4294
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     promoter        complement(4564..4582)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4603..4619)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4627..4643)
                     /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(4651..4681)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4696..4717)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5005..5593)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5767..6624)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6625..6729)
                     /label="AmpR promoter"
     rep_origin      complement(join(6756..6864,1..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 gcgtgagcgg cgagggcgag ggcgacgcca cctacggcaa
     1381 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctggt
     1441 gaccaccctg agctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     1501 cgacttcttc aagagcgcca tgcccgaggg ctacgtgcag gagcgcacca tcttcttcaa
     1561 ggacgacggt gagcttgcgg ggttgcgagc aacactccag caacgaacag tgcccaagtc
     1621 aggaatctgc agtcagcctg ggctttcggc ggctttttct tgggcaaaca gcttgcactc
     1681 atgccagcgc ggcttgtcca gcctcacttg agctttccag ctgctaccag ccgggctata
     1741 cgacagcgac agagccatag cgtggaatca cttatttggg ttgccgaagt agcggtcgga
     1801 gcgtgagttc ttggtcaagc cgccccttat ccggttcctg tccgtgtctt tgtccctcgt
     1861 tcacccttcg cggcaccctt catccccttg cttgcaggta actacaagac ccgcgccgag
     1921 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1981 gaggacggca acatcctggg ccacaagctg gagtacaacg ccatccacgg caacgtgtac
     2041 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcggcctgaa ctgcaacatc
     2101 gaggacggca gcgtgcagct ggccgaccac taccagcaga acacccccat cggcgacggc
     2161 cccgtgctgc tgcccgacaa ccactacctg agcacccaga gcaagctgag caaggacccc
     2221 aacgagaagc gcgaccacat ggtgctgctg gagttcgtga ccgccgccgg catcaccctg
     2281 ggcatggacg agctgtacaa gatcgagggc agggatatcg aattctggag ccacccgcag
     2341 ttcgagaagt aaccgctccg tgtaaatgga ggcgctcgtt gatctgagcc ttgccccctg
     2401 acgaacggcg gtggatggaa gatactgctc tcaagtgctg aagcggtagc ttagctcccc
     2461 gtttcgtgct gatcagtctt tttcaacacg taaaaagcgg aggagttttg caattttgtt
     2521 ggttgtaacg atcctccgtt gattttggcc tctttctcca tgggcgggct gggcgtattt
     2581 gaagcgggat ccttcgaacg tacggttaac tcgcgacctg caggtacgta atttaaatac
     2641 gcgtgctgag gcttgacatg attggtgcgt atgtttgtat gaagctacag gactgatttg
     2701 gcgggctatg agggcggggg aagctctgga agggccgcga tggggcgcgc ggcgtccaga
     2761 aggcgccata cggcccgctg gcggcaccca tccggtataa aagcccgcga ccccgaacgg
     2821 tgacctccac tttcagcgac aaacgagcac ttatacatac gcgactattc tgccgctata
     2881 cataaccact cagctagctt aagatcccat ccctagggca tgccgggcgc gccagaagga
     2941 gcgcagccaa accaggatga tgtttgatgg ggtatttgag cacttgcaac ccttatccgg
     3001 aagccccctg gcccacaaag gctaggcgcc aatgcaagca gttcgcatgc agcccctgga
     3061 gcggtgccct cctgataaac cggccagggg gcctatgttc tttacttttt tacaagagaa
     3121 gtcactcaac atcttaaaat ggccaggtga gtcgacgagc aagcccggcg gatcaggcag
     3181 cgtgcttgca gatttgactt gcaacgcccg cattgtgtcg acgaaggctt ttggctcctc
     3241 tgtcgctgtc tcaagcagca tctaaccctg cgtcgccgtt tccatttgca ggaagcttat
     3301 gacacaagaa tccctgttac ttctcgaccg tattgattcg gatgattcct acgcgagcct
     3361 gcggaacgac caggagttct gggagccgct ggcccgccga gccctggagg agctcgggct
     3421 gccggtgccg ccggtgctgc gggtgcccgg cgagagcacc aaccccgtac tggtcggcga
     3481 gcccggcccg gtgatcaagc tgttcggcga gcactggtgc ggtccggaga gcctcgcgtc
     3541 ggagtcggag gcctacgcgg tcctggcgga cgccccggtg ccggtgcccc gcctcctcgg
     3601 ccgcggcgag ctgcggcccg gcaccggagc ctggccgtgg ccctacctgg tgatgagccg
     3661 gatgaccggc accacctggc ggtccgcgat ggacggcacg accgaccgga acgcgctgct
     3721 cgccctggcc cgcgaactcg gccgggtgct cggccggctg cacagggtgc cgctgaccgg
     3781 gaacaccgtg ctcacccccc attccgaggt cttcccggaa ctgctgcggg aacgccgcgc
     3841 ggcgaccgtc gaggaccacc gcgggtgggg ctacctctcg ccccggctgc tggaccgcct
     3901 ggaggactgg ctgccggacg tggacacgct gctggccggc cgcgaacccc ggttcgtcca
     3961 cggcgacctg cacgggacca acatcttcgt ggacctggcc gcgaccgagg tcaccgggat
     4021 cgtcgacttc accgacgtgt atgcgggaga ctcccgctac agcctggtgc aactgcatct
     4081 caacgccttc cggggcgacc gcgagatcct ggccgcgctg ctcgacgggg cgcagtggaa
     4141 gcggaccgag gacttcgccc gcgaactgct cgccttcacc ttcctgcacg acttcgaggt
     4201 gttcgaggag accccgctgg atctctccgg cttcaccgat ccggaggaac tggcgcagtt
     4261 cctctggggg ccgccggaca ccgcccccgg cgcctgactc gagtgaccgc tccgtgtaaa
     4321 tggaggcgct cgttgatctg agccttgccc cctgacgaac ggcggtggat ggaagatact
     4381 gctctcaagt gctgaagcgg tagcttagct ccccgtttcg tgctgatcag tctttttcaa
     4441 cacgtaaaaa gcggaggagt tttgcaattt tgttggttgt aacgatcctc cgttgatttt
     4501 ggcctctttc tccatgggcg ggctgggcgt atttgaagcg gacccggtac ccagcttttg
     4561 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     4621 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4681 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4741 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4801 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4861 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4921 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4981 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5041 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5101 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5161 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5221 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5281 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5341 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5401 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     5461 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5521 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5581 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5641 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5701 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5761 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5821 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5881 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5941 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6001 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6061 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6121 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6181 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6241 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6301 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6361 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6421 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6481 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6541 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6601 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6661 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6721 ggttccgcgc acatttcccc gaaaagtgcc acactaaatt gtaagcgtta atattttgtt
     6781 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     6841 caaaatccct tataaatcaa aaga
//
