LOCUS       Exported                3860 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSTn5-KM, complete sequence.
ACCESSION   KC989926
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3860)
  AUTHORS   Madhaiyan M, Peng N, Ngoh ST, Cheng H, Cai L, Fu L, Reddy C, Hong Y,
            Ji L.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-APR-2013) Biomaterials & Biocatalysts Group, Temasek 
            Life Sciences Laboratory, 1 Research Link, National University of 
            Singapore, Singapore 117604, Singapore
REFERENCE   2  (bases 1 to 3860)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3860
                     /organism="Cloning vector pSTn5-KM"
                     /mol_type="other DNA"
                     /db_xref="taxon:1389801"
     misc_feature    25..58
                     /label=loxP-RE
                     /note="loxP-RE"
     protein_bind    25..58
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      59..143
                     /regulatory_class="promoter"
                     /note="derived from mdh promoter in Methylobacterium 
                     radiotolerans"
     regulatory      144..191
                     /regulatory_class="promoter"
                     /note="T5 promoter"
     promoter        147..191
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    167..183
                     /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)."
     CDS             214..1008
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /note="aphA with Methylobacterium radiotolerans codon 
                     preference"
                     /protein_id="AGV01072.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     RBS             1016..1024
                     /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)"
     terminator      1043..1090
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      1044..1090
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     misc_feature    1091..1124
                     /label=loxP-LE
                     /note="loxP-LE"
     protein_bind    1091..1124
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(1210..1226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1234..1250
                     /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)."
     promoter        complement(1258..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1303..1324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1612..2200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2371..3231)
                     /codon_start=1
                     /product="ampicilin resistance protein"
                     /label=ampicilin resistance protein
                     /note="Amp"
                     /protein_id="AGV01073.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3232..3336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3810..3826
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3841..3859
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"