LOCUS       Exported               10456 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPBAsyn, complete sequence.
ACCESSION   KX573871
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10456)
  AUTHORS   Ghosh IN, Landick R.
  TITLE     OptSSeq: High-throughput sequencing readout of growth enrichment 
            defines optimal gene expression elements for homoethanologenesis
  JOURNAL   ACS Synth Biol (2016) In press
  PUBMED    27404024
REFERENCE   2  (bases 1 to 10456)
  AUTHORS   Ghosh IN, Landick RC.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2016) DOE Great Lakes Bioenergy Research Center, 
            University of Wisconsin-Madison, 1552 University Avenue, Madison, WI
            53726, United States
REFERENCE   3  (bases 1 to 10456)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10456
                     /organism="Cloning vector pPBAsyn"
                     /mol_type="other DNA"
                     /db_xref="taxon:1881972"
     regulatory      35..96
                     /regulatory_class="promoter"
                     /function="Stationary phase promoter"
                     /note="Dps100"
     regulatory      108..181
                     /regulatory_class="promoter"
                     /function="Expressed in anaerobic conditions"
                     /note="YdfZ promoter"
     CDS             221..1927
                     /codon_start=1
                     /product="Pdc"
                     /label=Pdc
                     /note="Pyruvate Decarboxylase (recoded)"
                     /protein_id="AOH95439.1"
                     /translation="MSYTVGTYLAERLVQIGLKHHFAVAGDYNLVLLDNLLLNKNMEQV
                     YCCNELNCGFSAEGYARAKGAAAAVVTYSVGALSAFDAIGGAYAENLPVILISGAPNNN
                     DHAAGHVLHHALGKTDYHYQLEMAKNITAAAEAIYTPEEAPAKIDHVIKTALREKKPVY
                     LEIACNIASMPCAAPGPASALFNDEASDEASLNAAVEETLKFIANRDKVAVLVGSKLRA
                     AGAEEAAVKFADALGGAVATMAAAKSFFPEENPHYIGTSWGEVSYPGVEKTMKEADAVI
                     ALAPVFNDYSTTGWTDIPDPKKLVLAEPRSVVVNGVRFPSVHLKDYLTRLAQKVSKKTG
                     ALDFFKSLNAGELKKAAPADPSAPLVNAEIARQVEALLTPNTTVIAETGDSWFNAQRMK
                     LPNGARVEYEMQWGHIGWSVPAAFGYAVGAPERRNILMVGDGSFQLTAQEVAQMVRLKL
                     PVIIFLINNYGYTIEVMIHDGPYNNIKNWDYAGLMEVFNGNGGYDSGAGKGLKAKTGGE
                     LAEAIKVALANTDGPTLIECFIGREDCTEELVKWGKRVAAANSRKPVNKLL"
     CDS             1987..3138
                     /codon_start=1
                     /product="AdhB"
                     /label=AdhB
                     /note="Alcohol Dehydrogenase II (Recoded)"
                     /protein_id="AOH95440.1"
                     /translation="MASSTFYIPFVNEMGEGSLEKAIKDLNGSGFKNALIVSDAFMNKS
                     GVVKQVADLLKAQGINSAVYDGVMPNPTVTAVLEGLKILKDNNSDFVISLGGGSPHDCA
                     KAIALVATNGGEVKDYEGIDKSKKPALPLMSINTTAGTASEMTRFCIITDEVRHVKMAI
                     VDRHVTPMVSVNDPLLMVGMPKGLTAATGMDALTHAFEAYSSTAATPITDACALKAASM
                     IAKNLKTACDNGKDMPAREAMAYAQFLAGMAFNNASLGYVHAMAHQLGGYYNLPHGVCN
                     AVLLPHVLAYNASVVAGRLKDVGVAMGLDIANLGDKEGAEATIQAVRDLAASIGIPANL
                     TELGAKKEDVPLLADHALKDACALTNPRQGDQKEVEELFLSAF"
     CDS             3167..4180
                     /codon_start=1
                     /product="AdhA"
                     /label=AdhA
                     /note="Alcohol Dehydrogenase I (Recoded)"
                     /protein_id="AOH95441.1"
                     /translation="MKAAVITKDHTIEVKDTKLRPLKYGEALLEMEYCGVCHTDLHVKN
                     GDFGDETGRITGHEGIGIVKQVGEGVTSLKVGDRASVAWFFKGCGHCEYCVSGNETLCR
                     NVENAGYTVDGAMAEECIVVADYSVKVPDGLDPAVASSITCAGVTTYKAVKVSQIQPGQ
                     WLAIYGLGGLGNLALQYAKNVFNAKVIAIDVNDEQLAFAKELGADMVINPKNEDAAKII
                     QEKVGGAHATVVTAVAKSAFNSAVEAIRAGGRVVAVGLPPEKMDLSIPRLVLDGIEVLG
                     SLVGTREDLKEAFQFAAEGKVKPKVTKRKVEEINQIFDEMEHGKFTGRMVVDFTHH"
     CDS             4211..4996
                     /codon_start=1
                     /product="Evoglow"
                     /label=Evoglow
                     /protein_id="AOH95442.1"
                     /translation="MASFQSFGIPGQLEVIKKALDHVRVGVVITDPALEDNPIVYVNQG
                     FVQMTGYETEEILGKNARFLQGKHTDPAEVDNIRTALQNKEPVTVQIQNYKKDGTMFWN
                     ELNIDPMEIEDKTYFVGIQNDITKQKEYEKLLEDSLTEITALSTPIVPIRNGISALPLV
                     GNLTEERFNSIVCTLTNILSTSKDDYLIIDLSGLAQVNEQTADQIFKLSHLLKLTGTEL
                     IITGIKPELAMKMNKLDANFSSLKTYSNVKDAVKVLPIM"
     protein_bind    complement(5001..5034)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             5153..5968
                     /codon_start=1
                     /product="AphA"
                     /label=AphA
                     /note="Aminoglycoside-3-phosphotransferase (Kanamycin 
                     resistance)"
                     /protein_id="AOH95443.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     protein_bind    complement(6062..6095)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      6108..6159
                     /regulatory_class="terminator"
                     /note="P22 terminator"
     promoter        6232..6260
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6449..6982
                     /codon_start=1
                     /product="AacC1"
                     /label=AacC1
                     /note="Gentamycon-3-acetyltransferase (Inactive)"
                     /protein_id="AOH95444.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     misc_feature    complement(7037..8434)
                     /label=Mob element
                     /note="Mob element"
     rep_origin      8265..9034
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             9023..9697
                     /codon_start=1
                     /product="REP"
                     /label=REP
                     /protein_id="AOH95445.1"
                     /translation="MEISMATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAA
                     MLLHHLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVS
                     AYVVNDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTG
                     PGEEPPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLP
                     RR"
     CDS             9035..9697
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     10404..10420
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        10430..10448
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"