LOCUS       40924_33822        6671 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pOpt_Clover_Paro, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6671)
  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  JOURNAL   Unpublished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 6671)
  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 6671)
  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 6671)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6671)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     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"
COMMENT     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"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            On May 18, 2016 this sequence version replaced KM061062.1.
FEATURES             Location/Qualifiers
     source          1..6671
                     /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
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             2269..2280
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             2293..2316
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     promoter        complement(4371..4389)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4410..4426)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4434..4450)
                     /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(4458..4488)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4503..4524)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4812..5400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5574..6431)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6432..6536)
                     /label=AmpR promoter
     rep_origin      complement(join(6563..6671,1..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 agcttactcc gccctccccg gtgctgaaga atttcgaagc
     3301 atggacgatg cgttgcgtgc actgcggggt cggtatcccg gttgtgagtg ggttgttgtg
     3361 gaggatgggg cctcgggggc tggtgtttat cggcttcggg gtggtgggcg ggagttgttt
     3421 gtcaaggtgg cagctctggg ggccggggtg ggcttgttgg gtgaggctga gcggctggtg
     3481 tggttggcgg aggtggggat tcccgtacct cgtgttgtgg agggtggtgg ggacgagagg
     3541 gtcgcctggt tggtcaccga agcggttccg gggcgtccgg ccagtgcgcg gtggccgcgg
     3601 gagcagcggc tggacgtggc ggtggcgctc gcggggctcg ctcgttcgct gcacgcgctg
     3661 gactgggagc ggtgtccgtt cgatcgcagt ctcgcggtga cggtgccgca ggcggcccgt
     3721 gctgtcgctg aagggagcgt cgacttggag gatctggacg aggagcggaa ggggtggtcg
     3781 ggggagcggc ttctcgccga gctggagcgg actcggcctg cggacgagga tctggcggtt
     3841 tgccacggtc acctgtgccc ggacaacgtg ctgctcgacc ctcgtacctg cgaggtgacc
     3901 gggctgatcg acgtggggcg ggtcggccgt gcggaccggc actccgatct cgcgctggtg
     3961 ctgcgcgagc tggcccacga ggaggacccg tggttcgggc cggagtgttc cgcggcgttc
     4021 ctgcgggagt acgggcgcgg gtgggatggg gcggtatcgg aggaaaagct ggcgttttac
     4081 cggctgttgg acgagttctt ctgactcgag tgaccgctcc gtgtaaatgg aggcgctcgt
     4141 tgatctgagc cttgccccct gacgaacggc ggtggatgga agatactgct ctcaagtgct
     4201 gaagcggtag cttagctccc cgtttcgtgc tgatcagtct ttttcaacac gtaaaaagcg
     4261 gaggagtttt gcaattttgt tggttgtaac gatcctccgt tgattttggc ctctttctcc
     4321 atgggcgggc tgggcgtatt tgaagcggac ccggtaccca gcttttgttc cctttagtga
     4381 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4441 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4501 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4561 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4621 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4681 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4741 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4801 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4861 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4921 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4981 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5041 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5101 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5161 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5221 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5281 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5341 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5401 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5461 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5521 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5581 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5641 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5701 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5761 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5821 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5881 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5941 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6001 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6061 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6121 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6181 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6241 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6301 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6361 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6421 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6481 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6541 tttccccgaa aagtgccaca ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     6601 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     6661 aaatcaaaag a
//
