LOCUS       40924_37373        8933 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Protein fusion vector pRM-vgb, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8933)
  AUTHORS   Mora-Lugo R, Madrigal M, Yelemane V, Fernandez-Lahore M.
  TITLE     Improved biomass and protein production in solid-state cultures of 
            an Aspergillus sojae strain harboring the Vitreoscilla hemoglobin
  JOURNAL   Appl. Microbiol. Biotechnol. 99 (22), 9699-9708 (2015)
  PUBMED    26224427
REFERENCE   2  (bases 1 to 8933)
  AUTHORS   Mora-Lugo R, Madrigal M, Yelemane V, Fernandez-Lahore M.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JUN-2015) School of Engineering and Science, Jacobs 
            University Bremen gGmbH, Campus Ring 1, Bremen, Bremen 28759, 
            Germany
REFERENCE   3  (bases 1 to 8933)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8933)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol."; date: "2015"; volume: "99"; issue: "22"; 
            pages: "9699-9708"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-JUN-2015) School of Engineering and Science, Jacobs University 
            Bremen gGmbH, Campus Ring 1, Bremen, Bremen 28759, 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..8933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1177..1292
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     CDS             1802..2518
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2554..2570)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(2806..2830)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(2959..4104)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(4406..5197)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      complement(5479..6110)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    complement(6249..6273)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(6389..6956)
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus
                     nidulans trpC gene"
     CDS             complement(7115..7486)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(7493..7853)
                     /label=trpC promoter
                     /note="promoter for Aspergillus nidulans trpC"
     promoter        7933..8933
                     /label=gpdA promoter
                     /note="promoter from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
ORIGIN
        1 accaaaagtc acacaacaca agctgtaagg atttcggcac ggctacggaa gacggagaag
       61 ccaccttcag tggactcgag taccatttaa ttctatttgt gtttgatcga gacctaatac
      121 agcccctaca acgaccatca aagtcgtata gctaccagtg aggaagtgga ctcaaatcga
      181 cttcagcaac atctcctgga taaactttaa gcctaaacta tacagaataa gataggtgga
      241 gagcttatac cgagctccca aatctgtcca gatcatggtt gaccggtgcc tggatcttcc
      301 tatagaatca tccttattcg ttgacctagc tgattctgga gtgacccaga gggtcatgac
      361 ttgagcctaa aatccgccgc ctccaccatt tgtagaaaaa tgtgacgaac tcgtgagctc
      421 tgtacagtga ccggtgactc tttctggcat gcggagagac ggacggacgc agagagaagg
      481 gctgagtaat aagccactgg ccagacagct ctggcggctc tgaggtgcag tggatgatta
      541 ttaatccggg accggccgcc cctccgcccc gaagtggaaa ggctggtgtg cccctcgttg
      601 accaagaatc tattgcatca tcggagaata tggagcttca tcgaatcacc ggcagtaagc
      661 gaaggagaat gtgaagccag gggtgtatag ccgtcggcga aatagcatgc cattaaccta
      721 ggtacagaag tccaattgct tccgatctgg taaaagattc acgagatagt accttctccg
      781 aagtaggtag agcgagtacc cggcgcgtaa gctccctaat tggcccatcc ggcatctgta
      841 gggcgtccaa atatcgtgcc tctcctgctt tgcccggtgt atgaaaccgg aaaggccgct
      901 caggagctgg ccagcggcgc agaccgggaa cacaagctgg cagtcgaccc atccggtgct
      961 ctgcactcga cctgctgagg tccctcagtc cctggtaggc agctttgccc cgtctgtccg
     1021 cccggtgtgt cggcggggtt gacaaggtcg ttgcgtcagt ccaacatttg ttgccatatt
     1081 ttcctgctct ccccaccagt tgctcttttc ttttctcttt cttttcccat cttcagtata
     1141 ttcatcttcc catccaagaa cctttatttc ccctaagtaa gtactttgct acatccatac
     1201 tccatccttc ccatccctta ttcctttgaa cctttcagtt cgagctttcc cacttcatcg
     1261 cagcttgact aacagctacc ccgcttgagc agacatcacc ctgggctgag gacttaacta
     1321 gtgacgcaag gtcatgctcg accagcagac catcaacatc atcaaggcca ccgtccccgt
     1381 cctcaaggag cacggtgtca ccatcaccac caccttctac aagaacctct tcgccaagca
     1441 ccccgaggtc cgccccctct tcgacatggg tcgccaggag tccctcgagc agcccaaggc
     1501 cctcgccatg accgtcctcg ccgccgccca gaacatcgag aacctccccg ccatcctccc
     1561 cgccgtcaag aagatcgccg tcaagcactg ccaggccggt gtcgccgccg cccactaccc
     1621 catcgtcggt caggagctgc tcggtgccat caaggaggtc ctcggtgacg ccgccaccga
     1681 cgacatcctc gacgcctggg gtaaggccta cggtgtcatc gccgacgtct tcatccaggt
     1741 cgaggccgac ctctacgccc aggccgtcga ggaccttgcg tcgaagcttg gtacccagct
     1801 gatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1861 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1921 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1981 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2041 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2101 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2161 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2221 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2281 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2341 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2401 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2461 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2521 aattaaaccc tcagcggtac cgagctctaa ttaactggcc gtcgttttac aacgtcgtga
     2581 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     2641 ttggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     2701 tggcgaatgg cgccgctgtt gcccgtctca ctggtgaaaa gaaaaaccac cccagtacat
     2761 taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc aatttgttta caccacaata
     2821 tatcctgcca ccagccagcc aacagctccc cgaccggcag ctcggcacaa aatcaccact
     2881 cgatacaggc agcccatcag tccgggacgg cgtcagcggg agagcagatc cagccggaac
     2941 tgaccccaca aggccctagc gtttgcaatg caccaggtca tcattgaccc aggcgtgttc
     3001 caccaggccg ctgcctcgca actcttcgca ggcttcgccg acctgctcgc gccacttctt
     3061 cacgcgggtg gaatccgatc cgcacatgag gcggaaggtt tccagcttga gcgggtacgg
     3121 ctcccggtgc gagctgaaat agtcgaacat ccgtcgggcc gtcggcgaca gcttgcggta
     3181 cttctcccat atgaatttcg tgtagtggtc gccagcaaac agcacgacga tttcctcgtc
     3241 gatcaggacc tggcaacggg acgttttctt gccacggtcc aggacgcgga agcggtgcag
     3301 cagcgacacc gattccaggt gcccaacgcg gtcggacgtg aagcccatcg ccgtcgcctg
     3361 taggcgcgac aggcattcct cggccttcgt gtaataccgg ccattgatcg accagcccag
     3421 gtcctggcaa agctcgtaga acgtgaaggt gatcggctcg ccgatagggg tgcgcttcgc
     3481 gtactccaac acctgctgcc acaccagttc gtcatcgtcg gcccgcagct cgacgccggt
     3541 gtaggtgatc ttcacgtcct tgttgacgtg gaaaatgacc ttgttttgca gcgcctcgcg
     3601 cgggattttc ttgttgcgcg tggtgaacag ggcagagcgg gccgtgtcgt ttggcatcgc
     3661 tcgcatcgtg tccggccacg gcgcaatatc gaacaaggaa agctgcattt ccttgatctg
     3721 ctgcttcgtg tgtttcagca acgcggcctg cttggcctcg ctgacctgtt ttgccaggtc
     3781 ctcgccggcg gtttttcgct tcttggtcgt catagttcct cgcgtgtcga tggtcatcga
     3841 cttcgccaaa cctgccgcct cctgttcgag acgacgcgaa cgctccacgg cggccgatgg
     3901 cgcgggcagg gcagggggag ccagttgcac gctgtcgcgc tcgatcttgg ccgtagcttg
     3961 ctggaccatc gagccgacgg actggaaggt ttcgcggggc gcacgcatga cggtgcggct
     4021 tgcgatggtt tcggcatcct cggcggaaaa ccccgcgtcg atcagttctt gcctgtatgc
     4081 cttccggtca aacgtccgat tcattcaccc tccttgcggg attgccccga ctcacgccgg
     4141 ggcaatgtgc ccttattcct gatttgaccc gcctggtgcc ttggtgtcca gataatccac
     4201 cttatcggca atgaagtcgg tcccgtagac cgtctggccg tccttctcgt acttggtatt
     4261 ccgaatcttg ccctgcacga ataccagcga ccccttgccc aaatacttgc cgtgggcctc
     4321 ggcctgagag ccaaaacact tgatgcggaa gaagtcggtg cgctcctgct tgtcgccggc
     4381 atcgttgcgc cacatctagg tactaaaaca attcatccag taaaatataa tattttattt
     4441 tctcccaatc aggcttgatc cccagtaagt caaaaaatag ctcgacatac tgttcttccc
     4501 cgatatcctc cctgatcgac cggacgcaga aggcaatgtc ataccacttg tccgccctgc
     4561 cgcttctccc aagatcaata aagccactta ctttgccatc tttcacaaag atgttgctgt
     4621 ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg cttttccgtc tttaaaaaat
     4681 catacagctc gcgcggatct ttaaatggag tgtcttcttc ccagttttcg caatccacat
     4741 cggccagatc gttattcagt aagtaatcca attcggctaa gcggctgtct aagctattcg
     4801 tatagggaca atccgatatg tcgatggagt gaaagagcct gatgcactcc gcatacagct
     4861 cgataatctt ttcagggctt tgttcatctt catactcttc cgagcaaagg acgccatcgg
     4921 cctcactcat gagcagattg ctccagccat catgccgttc aaagtgcagg acctttggaa
     4981 caggcagctt tccttccagc catagcatca tgtccttttc ccgttccaca tcataggtgg
     5041 tccctttata ccggctgtcc gtcattttta aatataggtt ttcattttct cccaccagct
     5101 tatatacctt agcaggagac attccttccg tatcttttac gcagcggtat ttttcgatca
     5161 gttttttcaa ttccggtgat attctcattt tagccattta ttatttcctt cctcttttct
     5221 acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt
     5281 ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt
     5341 ggcgtataac atagtatcga cggagccgat tttgaaacca caattatggg tgatgctgcc
     5401 aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggcgt acgcctgggg
     5461 ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg
     5521 gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg
     5581 gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc
     5641 aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc
     5701 tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc
     5761 ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga
     5821 aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg
     5881 gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag
     5941 tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac
     6001 gccggcggcc gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg tttgcagggc
     6061 catagacggc cgccagccca gcggcgaggg caaccagccc ggtgagcgtc gcaaaggcgc
     6121 tcggtcttgc cttgctcgtc ggatctggaa ccctgtggtt ggcatgaaca aacaaatgga
     6181 cgaacggata aaccttttca cgccctttta aatatccgat tattctaata aacgctcttt
     6241 tctcttaggt ttacccgcca atatatcctg tcaaacactg atagtttaaa ctgaaggcgg
     6301 gaaacgacaa tctgatcatg agcggagaat taagggagtc acgaagctaa atccgtttaa
     6361 acggcgcgcg gcgtagaaga tcctctagaa agaaggatta cctctaaaca agtgtacctg
     6421 tgcattctgg gtaaacgact cataggagag ttgtaaaaaa gtttcggccg gcgtattggg
     6481 tgttacggag cattcactag gcaaccatgg ttactattgt atacccatct tagtaggaat
     6541 gattttcgag gtttatacct acgatgaatg tgtgtcctgt aggcttgaga gttcaaggaa
     6601 gaaacagtgc aattatcttt gcgaacccag gggctggtga cggaattttc atagtcaagc
     6661 tatcagagta aagaagagga gcatgtcaaa gtacaattag agacaaatat atagtcgcgt
     6721 ggagccaaga gcggattcct cagtctcgta ggtctcttga cgaccgttga tctgcttgat
     6781 ctcgtctccc gaaaatgaaa atagactctg ctaagctatt cttctgcttc gccggagcct
     6841 gaagggcgta ctagggttgc gaggtccaat gcattaatgc attgcagatg agctgtatct
     6901 ggaagaggta aacccgaaac gcgttttatt cttgttgaca tggagctatt aaatcactag
     6961 aaggcactct ttgctgcttg gacaaatgaa cgtatcttat cgagatcctg aacaccattt
     7021 gtctcaactc cggagctgac atcgacacca acgatcttat atccagattc gtcaagctgt
     7081 ttgatgattt cagtaacgtt aagtggatcc gtcagtcctg ctcctcggcc acgaagtgca
     7141 cgcagttgcc ggccgggtcg cgcagggcga actcccgccc ccacggctgc tcgccgatct
     7201 cggtcatggc cggcccggag gcgtcccgga agttcgtgga cacgacctcc gaccactcgg
     7261 cgtacagctc gtccaggccg cgcacccaca cccaggccag ggtgttgtcc ggcaccacct
     7321 ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg gcgaagtcgt
     7381 cctccacgaa gtcccgggag aacccgagcc ggtcggtcca gaactcgacc gctccggcga
     7441 cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt ggccatggat cgatgcttgg
     7501 gtagaatagg taagtcagat tgaatctgaa ataaagggag gaagggcgaa cttaagaagg
     7561 tatgaccggg tcgttcactt accttgcttg acaaacgcac caagttatcg tgcaccaagc
     7621 agcagatgat aataatgtcc tcgttcctgt ctgctaataa gagtcacact tcgagcgccg
     7681 ccgctactgc tacaagtggg gctgatctga ccagttgcct aaatgaacca tcttgtcaaa
     7741 cgacacaaat tttgtgctca ccgcctggac gactaaacca aaataggcat tcattgttga
     7801 cctccactag ctccagccaa gcccaaaaag tgctccttca atatcatctt ctgtcgaggc
     7861 ctgatcatcg aagggcccat cgaagatcag gcctcgatat ctagagcgcg cctaggcggc
     7921 cgtggccagc ccgaattccc ttgtatctct acacacaggc tcaaatcaat aagaagaacg
     7981 gttcgtcttt ttcgtttata tcttgcatcg tcccaaagct attggcggga tattctgttt
     8041 gcagttggct gacttgaagt aatctctgca gatctttcga cactgaaata cgtcgagcct
     8101 gctccgcttg gaagcggcga ggagcctcgt cctgtcacaa ctaccaacat ggagtacgat
     8161 aagggccagt tccgccagct cattaagagc cagttcatgg gcgttggcat gatggccgtc
     8221 atgcatctgt acttcaagta caccaacgct cttctgatcc agtcgatcat ccgctgaagg
     8281 cgctttcgaa tctggttaag atccacgtct tcgggaagcc agcgactggt gacctccagc
     8341 gtccctttaa ggctgccaac agctttctca gccagggcca gcccaagacc gacaaggcct
     8401 ccctccagaa cgccgagaag aactggaggg gtggtgtcaa ggaggagtaa gctccttatt
     8461 gaagtcggag gacggagcgg tgtcaagagg atattcttcg actctgtatt atagataaga
     8521 tgatgaggaa ttggaggtag catagcttca tttggatttg ctttccaggc tgagactcta
     8581 gcttggagca tagagggtcc tttggctttc aatattctca agtatctcga gtttgaactt
     8641 attccctgtg aaccttttat tcaccaatga gcattggaat gaacatgaat ctgaggactg
     8701 caatcgccat gaggttttcg aaatacatcc ggatgtcgaa ggcttggggc acctgcgttg
     8761 gttgaattta gaacgtggca ctattgatca tccgatagct ctgcaaaggg cgttgcacaa
     8821 tgcaagtcaa acgttgctag cagttccagg tggaatgtta tgatgagcat tgtattaaat
     8881 caggagatat agcatgatct ctagttagct caccacaaaa gtcagacggc gta
//
