LOCUS       Exported                8662 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAMG258, complete sequence.
ACCESSION   JN880475
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8662)
  AUTHORS   Olson DG.
  TITLE     Transformation of Clostridium thermocellum by electroporation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8662)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2011) Thayer School of Engineering, Dartmouth 
            College, 8000 Cummings, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8662)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8662
                     /organism="Cloning vector pAMG258"
                     /mol_type="other DNA"
                     /label=for transformation of Clostridium thermocellum
                     /note="for transformation of Clostridium thermocellum"
                     /db_xref="taxon:1155008"
     rep_origin      1..519
                     /label=CEN6/ARSH
                     /note="CEN6/ARSH"
     misc_feature    11..514
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     regulatory      692..1339
                     /regulatory_class="promoter"
                     /gene="GapDH"
     gene            692..1339
                     /gene="GapDH"
                     /label=GapDH
     CDS             1340..1990
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="AFC37772.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            1340..1990
                     /gene="cat"
                     /label=cat
     CDS             2008..2553
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label=hpt
                     /protein_id="AFC37773.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            2008..2553
                     /gene="hpt"
                     /label=hpt
     terminator      2663..2910
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      3099..3772
                     /label=eco ori
                     /note="eco ori"
     rep_origin      complement(3158..3746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3833..4411)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label=tdk
                     /protein_id="AFC37774.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(3833..4411)
                     /gene="tdk"
                     /label=tdk
     regulatory      complement(4412..5032)
                     /regulatory_class="promoter"
                     /gene="Cbp"
     gene            complement(4412..5032)
                     /gene="Cbp"
                     /label=Cbp
     CDS             complement(5108..6112)
                     /codon_start=1
                     /gene="repB"
                     /product="RepB"
                     /label=repB
                     /protein_id="AFC37775.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(5108..6112)
                     /gene="repB"
                     /label=repB
     rep_origin      complement(6113..6567)
                     /direction=LEFT
                     /label=cth ori
                     /note="cth ori"
     CDS             complement(6670..7530)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AFC37776.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6670..7530)
                     /gene="bla"
                     /label=bla
     CDS             complement(7626..8429)
                     /codon_start=1
                     /product="URA3"
                     /label=URA3
                     /protein_id="AFC37777.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(8430..8645)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter