LOCUS       40924_33842        6704 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pOpt_mCerulean3_Paro, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6704)
  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  JOURNAL   Unpublished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 6704)
  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 6704)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6704)
  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            "Unpublished"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"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6704
                     /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             2302..2313
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             2326..2349
                     /codon_start=1
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
                     /translation="WSHPQFEK"
     promoter        complement(4404..4422)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4443..4459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4467..4483)
                     /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(4491..4521)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4536..4557)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4845..5433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5607..6464)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6465..6569)
                     /label=AmpR promoter
     rep_origin      complement(join(6596..6704,1..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 ggaagcttac
     3301 tccgccctcc ccggtgctga agaatttcga agcatggacg atgcgttgcg tgcactgcgg
     3361 ggtcggtatc ccggttgtga gtgggttgtt gtggaggatg gggcctcggg ggctggtgtt
     3421 tatcggcttc ggggtggtgg gcgggagttg tttgtcaagg tggcagctct gggggccggg
     3481 gtgggcttgt tgggtgaggc tgagcggctg gtgtggttgg cggaggtggg gattcccgta
     3541 cctcgtgttg tggagggtgg tggggacgag agggtcgcct ggttggtcac cgaagcggtt
     3601 ccggggcgtc cggccagtgc gcggtggccg cgggagcagc ggctggacgt ggcggtggcg
     3661 ctcgcggggc tcgctcgttc gctgcacgcg ctggactggg agcggtgtcc gttcgatcgc
     3721 agtctcgcgg tgacggtgcc gcaggcggcc cgtgctgtcg ctgaagggag cgtcgacttg
     3781 gaggatctgg acgaggagcg gaaggggtgg tcgggggagc ggcttctcgc cgagctggag
     3841 cggactcggc ctgcggacga ggatctggcg gtttgccacg gtcacctgtg cccggacaac
     3901 gtgctgctcg accctcgtac ctgcgaggtg accgggctga tcgacgtggg gcgggtcggc
     3961 cgtgcggacc ggcactccga tctcgcgctg gtgctgcgcg agctggccca cgaggaggac
     4021 ccgtggttcg ggccggagtg ttccgcggcg ttcctgcggg agtacgggcg cgggtgggat
     4081 ggggcggtat cggaggaaaa gctggcgttt taccggctgt tggacgagtt cttctgactc
     4141 gagtgaccgc tccgtgtaaa tggaggcgct cgttgatctg agccttgccc cctgacgaac
     4201 ggcggtggat ggaagatact gctctcaagt gctgaagcgg tagcttagct ccccgtttcg
     4261 tgctgatcag tctttttcaa cacgtaaaaa gcggaggagt tttgcaattt tgttggttgt
     4321 aacgatcctc cgttgatttt ggcctctttc tccatgggcg ggctgggcgt atttgaagcg
     4381 gacccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     4441 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4501 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4561 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4621 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4681 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4741 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4801 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4861 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4921 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4981 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5041 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5101 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5161 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5221 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5281 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5341 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5401 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5461 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5521 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5581 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5641 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5701 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5761 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5821 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5881 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5941 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6001 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6061 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6121 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6181 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6241 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6301 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6361 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6421 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6481 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6541 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acactaaatt
     6601 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     6661 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aaga
//
