LOCUS       Exported                5899 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCBcth8, complete sequence.
ACCESSION   MG026508
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5899)
  AUTHORS   Zheng T, Cui J, Bae HR, Lynd LR, Olson DG.
  TITLE     Expression of adhA from different organisms in Clostridium 
            thermocellum
  JOURNAL   Biotechnol Biofuels 10, 251 (2017)
  PUBMED    29213311
REFERENCE   2  (bases 1 to 5899)
  AUTHORS   Zheng T.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2017) Biology, Dartmouth College, 14 Engineering 
            Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 5899)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5899
                     /organism="Cloning vector pCBcth8"
                     /mol_type="other DNA"
                     /db_xref="taxon:2059363"
     regulatory      1..209
                     /regulatory_class="promoter"
                     /note="2638"
     CDS             210..1424
                     /codon_start=1
                     /gene="adhA"
                     /product="AdhA"
                     /label=adhA
                     /note="derived from Methanocaldococcus bathoardescens"
                     /protein_id="AUE21772.1"
                     /translation="MNYEKEVNINNVFELRCKTTDYFGVGAIHKFDDIAKEFVEKGMKN
                     FLFVTGKSSYKKCGAWDVIEPILDKYGINYELYNKVSANPTVDMIDEAVKLGKEIESQV
                     VIGIGGGSPLDTAKSVAVLLEYPDKTGRDLYEFKFAPERAKPIVAINTTHGTGSEVDRF
                     AVATVPEKNYKPAIVYDCIYPTYAIDDPALMTKLPKEQALATAIDAVNHVNEAATTKVA
                     SPYVVDLARRTVGLIKEYLPRVLENPEDLTARYWVLYASTIAGIALDNGLAHITHALEH
                     PLSAIDANVTHGIGLAALMPAVVKYCHPALPKTFAYIYEPIVEDKERAEKDANYLAQEI
                     EKWLFDLGLTQKLPDLGFKESDVDELTRLAMTTPSLDILLSLAPIEVNEDVVRNIYLDS
                     MQPIQ"
     gene            210..1424
                     /gene="adhA"
                     /label=adhA
     CDS             complement(2282..3142)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3143..3234)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3834..4379
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             4850..5854
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"