LOCUS       40924_24064        5325 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHA1001-ble-PeEGFP DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5325)
  AUTHORS   Abe J, Harayama S.
  TITLE     Expression of ble-PeEGFP fusion protein for bleomycin resistance 
            protein and codon-optimized green fluorescence protein in 
            Pseudococcomyxa sp. KJ
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5325)
  AUTHORS   Abe J, Harayama S.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-SEP-2016) Contact:Jun Abe Chuo University, Faculty of 
            Science and Engineering; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 112-8551, 
            Japan
REFERENCE   3  (bases 1 to 5325)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5325)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-SEP-2016) Contact:Jun Abe Chuo University, Faculty of Science 
            and Engineering; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 112-8551, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /label=AmpR promoter
     CDS             129..986
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1160..1748
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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)."
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2238..3027
                     /note="includes promoter region of Pseudococcomyxa
                     ellipsoidea nuclear gene RBCS for ribulose bisphosphate 
                     carboxylase/oxygenase small subunit"
                     /regulatory_class="promoter"
     misc_feature    3028..3399
                     /gene="ble-PeEGFP"
                     /note="ble; bleomycin resistance protein"
     misc_feature    3400..3417
                     /gene="ble-PeEGFP"
                     /label=linker
                     /note="linker"
     CDS             3418..4131
                     /label=EGFP
                     /note="enhanced GFP"
     regulatory      4141..4652
                     /note="includes terminator region of Pseudococcomyxa
                     ellipsoidea RBCS gene"
                     /regulatory_class="terminator"
     promoter        complement(4682..4700)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4710..4726)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4867..5322)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg
     2221 gccgctctag aactagtatg ccctcaatcg gccgatttca tgcatgaaga agaagatgat
     2281 gattacacat cattccagcc atctagggct gccatcagta attgcatgac ccgcaacttc
     2341 ccagtcagtc aaatgcacaa tagaagcata gttgcatggg aatcaaccca gcagcctccc
     2401 gcagagctag cctaggtacg tgtacgcagc cacataatca tcatgaatga ttctgcacgg
     2461 tgctatgata gcaacagata ctcgccagtg cgcacgcaag caatctcatc acaccactgt
     2521 ggtcaggcaa gcacgatata gatgtcaaat gacagctcca ttgggagtgc aagtctacta
     2581 gagtgcaagc ctactagtag taggaaggtc cagcgcatgc caaaactcct gctaaggggt
     2641 tgcccaagcc ccttggctgg ccaaaagtcc ttaatatttt gctaggtaaa ggggatcttc
     2701 ccggaggtgg tgttgatgaa catttgtaca ccagcatccg ggtccttttg gactctgaaa
     2761 acgtgagcga gcagcgactg cttgcagctt ccaaacgaaa ggtctggaag cgctggagtg
     2821 gtgagtcagg ggtgactcag caaacaccaa ggacacacgt tttcacccta ccctcagaac
     2881 ccttgaaata ggggtcgatt tccctctaac ttaagtggtt atccacaaac tcacgacgat
     2941 aagagccagc gccagcacca tttcgttcct cattttcagc tgaaagcaaa gcctttgcat
     3001 acttatcgct ctcagattac aatcgccatg gccaagctga ccagcgccgt tccggtgctc
     3061 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     3121 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     3181 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     3241 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     3301 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     3361 gccggcaact gcgtgcactt cgtggccgag gagcaggacg gcggctccgg cggccgcatg
     3421 gctaagggcg aggagctgtt caccggcgtg gtccccatcc tggtcgagct ggacggcgac
     3481 gtcaacggcc acaagttctc agtcagcggc gagggcgagg gcgacgctac ctacggcaag
     3541 ctgacactca agttcatctg caccacaggc aagctgcccg tgccctggcc caccctggtc
     3601 accacactca catacggcgt ccagtgcttc tcccgctacc ccgaccacat gaagcagcac
     3661 gacttcttca agagcgctat gcccgagggc tacgtgcagg agcgcaccat cttcttcaag
     3721 gacgacggca actacaagac cagggctgag gtgaagttcg agggcgacac actggtcaac
     3781 cgtatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg ccacaagctg
     3841 gagtacaact acaactcaca caacgtgtac atcatggccg acaagcagaa gaacggcatc
     3901 aaggtcaact tcaagatccg ccacaacatc gaggacggct ccgtgcagct ggctgaccac
     3961 taccagcaga acacccccat cggcgacggc cccgtgctgc tccccgacaa ccactacctg
     4021 tccacccaga gcgctctctc aaaggacccc aacgagaaga gggaccacat ggtgctgctg
     4081 gagttcgtga ccgctgctgg catcacactg ggcatggacg agctgtacaa gtaagtcgac
     4141 aggttctaag caatgacctg ggctgtacat cattagcaaa ctgaatctgc agctttctag
     4201 gctcttgctg ttcctaggaa cttggggttc aagactgggt ttttgctgtg gttcctgact
     4261 gcagaccatc actgtctggg ttgtcgtgtc taatgtccat aggtgtgagc acatgctcca
     4321 cttcaattcc tactgctggg taactggcgc tgtctgaggg acttctgaga agaatgccga
     4381 gaagaacttg agcgatttgt aaggggatat tttgggtctg taccaaaggc ccagactggg
     4441 cttccagtgc atcgcacata tgcatctgca tgtccattca tacaatcaac ctgttgactg
     4501 catgacccag atcacaacct ccttgagttg gaagtagctt cttctacaga tagaaaatgg
     4561 tgcatgtcag ctgagggttg caggcgttta aaagtgcatc aagtttgcag aacacatggt
     4621 gattcatgac tgcatggaaa cagaccagca cactcgaggg ggggcccggt acccaattcg
     4681 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     4741 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4801 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4861 gaatggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4921 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4981 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5041 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     5101 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     5161 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     5221 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     5281 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcag
//
