LOCUS       Exported                4762 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  E.coli-Thermotoga shuttle vector pDH10, complete sequence.
ACCESSION   JN813374
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4762)
  AUTHORS   Han D, Norris SM, Xu Z.
  TITLE     Construction and transformation of a Thermotoga-E. coli shuttle 
            vector
  JOURNAL   BMC Biotechnol. 12, 2 (2012)
  PUBMED    22225774
REFERENCE   2  (bases 1 to 4762)
  AUTHORS   Han D, Xu Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-OCT-2011) Department of Biological Sciences, Bowling 
            Green State University, 217 Life Sci. Bldg., Bowling Green, OH 
            43403, USA
REFERENCE   3  (bases 1 to 4762)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4762
                     /organism="Escherichia coli-Thermotoga shuttle vector
                     pDH10"
                     /mol_type="other DNA"
                     /db_xref="taxon:1127224"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             126..986
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AEW27250.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            126..986
                     /gene="bla"
                     /label=bla
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2069..2099
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2216..2232
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             2440..2448
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             2458..3219
                     /codon_start=1
                     /gene="kan"
                     /product="thermostable kanamycin adenyltransferase"
                     /label=kan
                     /protein_id="AEW27249.1"
                     /translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFYSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQKFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASLLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            2458..3219
                     /gene="kan"
                     /label=kan
     CDS             complement(3229..3867)
                     /codon_start=1
                     /gene="rep"
                     /product="putative Rep protein"
                     /label=rep
                     /protein_id="AEW27248.1"
                     /translation="MSKEIITQNRVEQGSLDSKCHKSPLASTRRKKRFYQRAVSGVTAG
                     KHRNEFIAFLTLTSSLESPADITHSWEKLKKRIRRRYGNFEYIWVRERTQSGLVHMHIL
                     FRGPYIPQDWISKNWEEIHKAKIVYVEAVWDTGKAIRYMMKYLSKEMEGRFGYSWKWIF
                     KGAAQVWKWLCRALRYEMKEIIKIWEKLIIEIPPEGIRWGRIWELVGYG"
     gene            complement(3229..3867)
                     /gene="rep"
                     /label=rep
     promoter        complement(4119..4137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4144..4160)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4302..4757
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"