LOCUS       Exported               10886 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration vector pSH016, complete sequence.
ACCESSION   KP245915
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10886)
  AUTHORS   Lo J, Zheng T, Hon S, Olson DG, Lynd LR.
  TITLE     The bifunctional alcohol and aldehyde dehydrogenase gene, adhE, is 
            necessary for ethanol production in Clostridium thermocellum and 
            Thermoanaerobacterium saccharolyticum
  JOURNAL   J. Bacteriol. 197 (8), 1386-1393 (2015)
  PUBMED    25666131
REFERENCE   2  (bases 1 to 10886)
  AUTHORS   Zheng T, Olson DG, Tian L, Bomble YJ, Himmel ME, Lo J, Hon S, Shaw 
            AJ, van Dijken JP, Lynd LR.
  TITLE     Cofactor Specificity of the Bifunctional Alcohol and Aldehyde 
            Dehydrogenase (AdhE) in Wild-Type and Mutant Clostridium 
            thermocellum and Thermoanaerobacterium saccharolyticum
  JOURNAL   J. Bacteriol. 197 (15), 2610-2619 (2015)
  PUBMED    26013492
REFERENCE   3  (bases 1 to 10886)
  AUTHORS   Hon S.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-DEC-2014) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   4  (bases 1 to 10886)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10886
                     /organism="Integration vector pSH016"
                     /mol_type="other DNA"
                     /db_xref="taxon:1615844"
     CDS             complement(558..1418)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="ampicillin resistance"
                     /protein_id="AJO16029.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(558..1418)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(1419..1523)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1613..2112
                     /label=5 homology flank #1
                     /note="5 homology flank #1"
     CDS             2691..3341
                     /codon_start=1
                     /gene="cat"
                     /product="Cat"
                     /label=cat
                     /note="chloramphenicol acetyltransferase"
                     /protein_id="AJO16030.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            2691..3341
                     /gene="cat"
                     /label=cat
     CDS             3359..3904
                     /codon_start=1
                     /gene="hpt"
                     /product="hpt"
                     /label=hpt
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="AJO16031.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            3359..3904
                     /gene="hpt"
                     /label=hpt
     misc_feature    4090..4589
                     /label=5 homology flank #2
                     /note="5 homology flank #2"
     misc_feature    6952..7451
                     /label=3 homology flank
                     /note="3 homology flank"
     rep_origin      complement(7932..8520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8607..9185)
                     /codon_start=1
                     /gene="tdk"
                     /product="Tdk"
                     /label=tdk
                     /note="thymidine kinase"
                     /protein_id="AJO16032.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(8607..9185)
                     /gene="tdk"
                     /label=tdk
     CDS             complement(9882..10886)
                     /codon_start=1
                     /gene="repB"
                     /product="repB"
                     /label=repB
                     /note="replication protein"
                     /protein_id="AJO16033.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(9882..10886)
                     /gene="repB"
                     /label=repB