LOCUS       40924_10461        8246 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Chloroplast expression vector pCEV4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8246)
  AUTHORS   Chen P-Y., To K-Y.
  TITLE     Construction of chloroplast expression vectors for higher plants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8246)
  AUTHORS   Chen P-Y., To K-Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-APR-2017) Agricultural Biotechnology Research Center, 
            Academia Sinica, No. 128, Academia Road, Section 2, Nankang, Taipei 
            11529, Taiwan
REFERENCE   3  (bases 1 to 8246)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8246)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-APR-2017) Agricultural Biotechnology Research Center, Academia 
            Sinica, No. 128, Academia Road, Section 2, Nankang, Taipei 11529, 
            Taiwan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    408..2396
                     /note="Nicotiana benthamiana left targeting region;
                     contains 16S rRNA and trnI"
     tRNA            join(1609..1645,2362..2396)
                     /product="tRNA-Ile"
     rep_origin      1826..1907
                     /note="chloroplast replication origin; oriA"
     regulatory      2403..2650
                     /note="atpI promoter; PatpI"
                     /regulatory_class="promoter"
     misc_feature    2651..2666
                     /label=cloning site
                     /note="cloning site"
     regulatory      2667..2835
                     /note="rps16 terminator; Trps16"
                     /regulatory_class="terminator"
     regulatory      2842..3060
                     /note="rrn promoter; Prrn"
                     /regulatory_class="promoter"
     CDS             3067..3855
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      3865..4086
                     /note="psbA terminator; TpsbA"
                     /regulatory_class="terminator"
     misc_feature    4093..5994
                     /note="Nicotiana benthamiana right targeting region;
                     contains trnA and 23S rRNA"
     tRNA            join(4156..4193,4903..4937)
                     /product="tRNA-Ala"
     primer_bind     complement(6025..6041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6049..6065)
                     /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(6073..6103)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6118..6139)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6427..7015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7189..8046)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8047..8151)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctctac caagctggag
      421 tacggtaggg gcagagggaa tttccggtgg agcggtgaaa tgcgtagaga tcggaaagaa
      481 caccaacggc gaaagcactc tgctgggccg acactgacac tgagagacga aagctagggg
      541 agcgaatggg attagatacc ccagtagtcc tagccgtaaa cgatggatac taggcgctgt
      601 gcgtatcgac ccgtgcagtg ctgtagctaa cgcgttaagt atcccgcctg gggagtacgt
      661 tcgcaagaat gaaactcaaa ggaattgacg ggggcccgca caagcggtgg agcatgtggt
      721 ttaattcgat gcaaagcgaa gaaccttacc agggcttgac atgccgcgaa tcctcttgaa
      781 agagaggggt gccttcggga acgcggacac aggtggtgca tggctgtcgt cagctcgtgc
      841 cgtaaggtgt tgggttaagt cccgcaacga gcgcaaccct cgtgtttagt tgccatcgtt
      901 gagtttggaa ccctgaacag actgccggtg ataagccgga ggaaggtgag gatgacgtca
      961 agtcatcatg ccccttatgc cctgggcgac acacgtgcta caatggccgg gacaaagggt
     1021 cgcgatcccg cgagggtgag ctaacctcaa aaacccgtcc tcagttcgga ttgcaggctg
     1081 caactcgcct gcatgaagcc ggaatcgcta gtaatcgccg gtcagccata cggcggtgaa
     1141 ttcgttcccg ggccttgtac acaccgcccg tcacactatg ggagctggcc atgcccgaag
     1201 tcgttacctt aaccgcaagg agggggatgc cgaaggcagg gctagtgact ggagtgaagt
     1261 cgtaacaagg tagccgtact ggaaggtgcg gctggatcac ctccttttca gggagagcta
     1321 atgcttgttg ggtattttgg tttgacactg cttcacaccc ccaaaaaaaa agaagggagc
     1381 tacgtctgag ttaaacttgg agatggaagt cttctttcct ttctcgacgg tgaagtaaga
     1441 ccaagctcat gagcttatta tcctaggtcg gaacaagttg ataggacccc cttttttacg
     1501 tccccatgtt ccccccgtgt ggcgacatgg gggcgaaaaa aggaaagaga gggatggggt
     1561 ttctctcgct tttggcatag cgggccccca gtgggaggct cgcacgacgg gctattagct
     1621 cagtggtaga gcgcgcccct gataattgcg tcgttgtgcc tgggctgtga gggctctcag
     1681 ccacatggat agttcaatgt gctcatcggc gcctgaccct gagatgtgga tcatccaagg
     1741 cacattagca tggcgtactc ctcctgttcg aaccggggtt tgaaaccaaa ctcctcctca
     1801 ggaggataga tggggcgatt cgggtgagat ccaatgtaga tccaactttc gattcactcg
     1861 tgggatccgg gcggtccggg ggggaccacc acggctcctc tcttctcgag aatccataca
     1921 tcccttatca gtgtatggac agctatctct cgagcacagg tttaggttcg gcctcaatgg
     1981 gaaaataaaa tggagcacct aacaacgcat cttcacagac caagaactac gagatcgccc
     2041 ctttcattct ggggtgacgg agggatcgta ccattcgagc cgtttttttc ttgactcgaa
     2101 atgggagcag gtttgaaaaa ggatcttaga gtgtctaggg ttgggccagg agggtctctt
     2161 aacgccttct tttttcttct catcggagtt atttcacaaa gacttgccag ggtaaggaag
     2221 aaggggggaa caagcacact tggagagcgc agtacaacgg agagttgtat gctgcgttcg
     2281 ggaaggatga atcgctcccg aaaaggaatc tattgattct ctcccaattg gttggaccgt
     2341 aggtgcgatg atttacttca cgggcgaggt ctctggttca agtccaggat ggcccagagc
     2401 tctctagcta tataagaaat ccttgattaa taataacata ataagataaa taacttactt
     2461 cagaaatccc ttatggaatc gcttactatt tctgaatttc aaaaaagaga taaaaatagc
     2521 tggggatatt atgtgattta ttagtattct aaatcttagt tggtattcaa aatatccgat
     2581 tcaagtagac aaagagatgg ttgaatcaaa aaattttgtt taaagttcaa ttttttcaga
     2641 gggcaaggca gcggccgcgg cgcgcctcaa ccgaaattca attaaggaaa taaattaagg
     2701 aaatacaaaa aggggggtag tcatttgtat ataactttgt atgacttttc tcttctattt
     2761 ttttgtattt cctccctttc cttttctatt tgtatttttt tatcattgct tccattgaat
     2821 tccgtgttct ataacggtac catcagttcg agcctgatta tccctaagcc caatgtgagt
     2881 ttttctagtt ggatttgctc ccccgccgtc gttcaatgag aatggataag aggctcgtgg
     2941 gattgacgtg agggggcagg gatggctata tttctgggag cgaactccgg gcgaatatga
     3001 agcgcatgga tacaagttat gccttggaat gaaagacaat tccgaatccg ctttgtctac
     3061 ggatccatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
     3121 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
     3181 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
     3241 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
     3301 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
     3361 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
     3421 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
     3481 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
     3541 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     3601 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     3661 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     3721 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     3781 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
     3841 aaggtagtcg gcaaataatc tagagatcct ggcctagtct ataggaggtt ttgaaaagaa
     3901 aggagcaata atcattttct tgttctatca agagggtgct attgctcctt tctttttttc
     3961 tttttattta tttactagta ttttacttac atagactttt ttgtttacat tatagaaaaa
     4021 gaaggagagg ttattttctt gcatttattc atgattgagt attctatttt gattttgtat
     4081 ttgtttctgc agctgcgcca gggaaaagaa tagaagaagc atctgactac ttcatgcatg
     4141 ctccacttgg ctcgggggga tatagctcag ttggtagagc tccgctcttg caattgggtc
     4201 gttgcgatta cgggttggat gtctaattgt ccaggcggta atgatagtat cttgtacctg
     4261 aaccggtggc tcactttttc taagtaatgg ggaagaggac cgaaacgtgc cactgaaaga
     4321 ctctactgag acaaagatgg gctgtcaaga acgtagagga ggtaggatgg gcagttggtc
     4381 agatctagta tggatcgtac atggacggta gttggagtcg gcggctctcc cagggttccc
     4441 tcatctgaga tctctgggga agaggatcaa gttggccctt gcgaacagct tgatgcacta
     4501 tctcccttca accctttgag cgaaatgcgg caaaagaaaa ggaaggaaaa tccatggacc
     4561 gaccccatca tctccacccc gtaggaacta cgagatcacc ccaaggacgc cttcggcatc
     4621 caggggtcac ggaccgacca tagaaccctg ttcaataagt ggaacgcatt agctgtccgc
     4681 tctcaggttg ggcagtcagg gtcggagaag ggcaatgact cattcttagt tagaatggga
     4741 ttccaactca gcaccttttg agtgagattt tgagaagagt tgctctttgg agagcacagt
     4801 acgatgaaag ttgtaagctg tgttcggggg ggagttattg tctatcgttg gcctctatgg
     4861 tagaatcagt cgggggacct gagaggcggt ggtttaccct gcggcggatg tcagcggttc
     4921 gagtccgctt atctccaact cgtgaactta gccgatacaa agctttatga tagcacccaa
     4981 tttttccgat tcggcggttc gatctatgat ttatcattca tggacgttga taagatccat
     5041 ccatttagca gcaccttagg atggcatagc cttaaaagtg aagggcgagg ttcaaacgag
     5101 gaaaggctta cggtggatac ctaggcaccc agagacgagg aagggcgtag taatcgacga
     5161 aatgcttcgg ggagttgaaa ataagcatag atccggagat tcccgaatag ggcaaccttt
     5221 cgaactgctg ctgaatccat gggcaggcaa gagacaacct ggcgaactga aacatcttag
     5281 tagccagagg aaaagaaagc aaaagcgatt cccgtagtag cggcgagcga aatgggagca
     5341 gcctaaaccg tgaaaacggg gttgtgggag agcaatacaa gcgtcgtgct gctaggcgaa
     5401 gcagcccgaa tgctgcaccc tagatggcga aagtccagta gccgaaagca tcactagctt
     5461 atgctctgac ccgagtagca tggggcacgt ggaatcccgt gtgaatcagc aaggaccacc
     5521 ttgcaaggct aaatactcct gggtgaccga tagcgaagta gtaccgtgag ggaagggtga
     5581 aaagaacccc catcggggag tgaaatagaa catgaaaccg taagctccca agcagtggga
     5641 ggagccaggg ctctgaccgc gtgcctgttg aagaatgagc cggcgactca taggcagtgg
     5701 cttggttaag ggaacccacc ggagccgtag cgaaagcgag tcttcatagg gcaattgtca
     5761 ctgcttatgg acccgaacct gggtgatcta tccatgacca ggatgaagct tgggtgaaac
     5821 taagtggagg tccgaaccga ctgatgttga agaatcagcg gatgagttgt ggttaggggt
     5881 gaaatgccac tcgaacccag agctagctgg ttctccccga aatgcgttga ggcgcagcag
     5941 ttgactggac atctaggggt aaagcactgt ttcggtgcgg gccgcgagag cggtctgcag
     6001 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6061 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6121 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6181 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8161 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8221 taggcgtatc acgaggccct ttcgtc
//
