LOCUS       62056_1955        6858 bp DNA     circular SYN 17-NOV-2021
DEFINITION  Expression vector p2BBAD-uspA, complete sequence.
ACCESSION   OL331257
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  AUTHORS   Tang R, Peng X, Weng C, Han Y.
  TITLE     The over-expression of phasin and regulator genes promoting the 
            synthesis of polyhydroxybutyrate in Cupriavidus necator H16 under 
            non-stress conditions
  JOURNAL   Appl Environ Microbiol, AEM0145821 (2021) In press
  PUBMED    34731058
REFERENCE   2  (bases 1 to 6858)
  AUTHORS   Tang R, Peng X, Weng C, Han Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-OCT-2021) National Key Laboratory of Biochemical 
            Engineering, Institute of Process Engineering, Chinese Academy of 
            Sciences, 1 North 2nd Street, Zhongguancun PR, Beijing, Beijing 
            100190, China
REFERENCE   3  (bases 1 to 6858)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol, AEM0145821 (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (31-OCT-2021) National Key Laboratory of Biochemical Engineering, 
            Institute of Process Engineering, Chinese Academy of Sciences, 1 
            North 2nd Street, Zhongguancun PR, Beijing, Beijing 100190, China"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2452
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(3248..4123)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        4150..4434
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      4493..4507
                     /regulatory_class="ribosome_binding_site"
     gene            4514..4948
                     /gene="uspA"
                     /label=uspA
     CDS             4514..4948
                     /codon_start=1
                     /transl_table=11
                     /gene="uspA"
                     /product="UspA"
                     /label=uspA
                     /note="universal stress protein, UspA family; derived from 
                     Cupriavidus necator H16"
                     /protein_id="UEP67417.1"
                     /translation="MFQHILLPTDGSELSKKAINGGLELAKAIGARITAYVCLEEYPYT
                     PFSEIVVEAPQAFKDRIENQAKLYLKEVENLATAAGVTFDADMSTFAVPYLGIIDAAER
                     HGCDVIFMASHGRRGLSGLLLGSETQKVLTHTDIPVIVYR"
     terminator      4959..5005
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        complement(5049..5067)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5088..5104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5112..5128)
                     /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(5136..5166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5181..5202)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5610..6401)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc
     1381 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg
     1441 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg
     1501 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag
     1561 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta
     1621 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg
     1681 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc
     1741 tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac
     1801 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
     1861 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
     1921 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
     1981 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     2041 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     2101 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     2161 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     2221 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     2281 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     2341 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     2401 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     2461 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     2521 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     2581 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga
     2641 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg
     2701 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     2761 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     2821 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag
     2881 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc
     2941 agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3001 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3061 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3121 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3181 gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc
     3241 tagattatga caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact
     3301 cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa
     3361 aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg
     3421 cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga
     3481 aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa
     3541 aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca
     3601 tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca
     3661 gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc
     3721 caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg
     3781 caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc
     3841 actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg
     3901 ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc
     3961 cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa
     4021 aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa
     4081 acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc
     4141 cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt
     4201 actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa
     4261 gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag
     4321 tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa
     4381 gattagcgga ttctacctga cgctttttat cgcaactctc tactgtttct ccatagggag
     4441 tccacaacgg tttccctcta gaaataattt tggaattcaa aagatctttt aagaaggaga
     4501 tatacataag cttatgttcc aacatatcct gctacccacc gacggctcgg aactctccaa
     4561 gaaagccatc aacggcgggc tggagctcgc caaggccatc ggcgcgcgca tcacggccta
     4621 tgtctgcctg gaggagtacc cctacacgcc gttcagcgaa atcgtggtcg aagccccgca
     4681 ggccttcaag gaccgcatcg agaaccaggc caagctgtat ctgaaagagg tcgagaacct
     4741 ggccacggcg gcgggcgtga ccttcgacgc cgatatgtcg acctttgcgg tgccctacct
     4801 gggcatcatc gacgcggccg agcgccatgg ctgcgacgtg atcttcatgg catctcatgg
     4861 ccggcgcggg ttgtccggcc tgctgctggg cagcgagacg cagaaggtgc tgacgcatac
     4921 cgacatcccg gtgatcgtct accgctgaaa gcttgcatca aataaaacga aaggctcagt
     4981 cgaaagactg ggcctttcgt tttatctgtg tcgacctcga gggggggccc ggtacccagc
     5041 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     5101 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5161 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5221 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5281 gggagaggcg gtttgcgtat tgggcgcatg cataaaaact gttgtaattc attaagcatt
     5341 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     5401 accttgtcgc cttgcgtata atatttgccc atgggggtgg gcgaagaact ccagcatgag
     5461 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     5521 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     5581 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     5641 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     5701 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     5761 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     5821 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     5881 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     5941 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     6001 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     6061 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     6121 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     6181 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     6241 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     6301 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     6361 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     6421 gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc
     6481 cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt
     6541 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     6601 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     6661 atcccaggtg gcacttttcg gggaaatgtg cgcgcccgcg ttcctgctgg cgctgggcct
     6721 gtttctggcg ctggacttcc cgctgttccg tcagcagctt ttcgcccacg gccttgatga
     6781 tcgcggcggc cttggcctgc atatcccgat tcaacggccc cagggcgtcc agaacgggct
     6841 tcaggcgctc ccgaaggt
//
