LOCUS       Exported                4730 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pYS1R1, complete sequence.
ACCESSION   HQ585611
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4730)
  AUTHORS   Shibayama Y, Dabbs ER, Yazawa K, Mikami Y.
  TITLE     Functional analysis of a small cryptic plasmid pYS1 from Nocardia
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4730)
  AUTHORS   Shibayama Y, Dabbs ER, Yazawa K, Mikami Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2010) School of Molecular and Cell Biology, 
            University of the Witwatersrand, 1 Jan Smuts Avenue, Johannesburg, 
            Gauteng 2050, South Africa
REFERENCE   3  (bases 1 to 4730)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4730
                     /organism="Shuttle vector pYS1R1"
                     /mol_type="other DNA"
                     /db_xref="taxon:915497"
     CDS             complement(431..1474)
                     /codon_start=1
                     /gene="rep"
                     /product="replication initiation protein"
                     /function="replication initiation in Gram-positive host"
                     /label=rep
                     /protein_id="ADR00353.1"
                     /translation="MKSCGWEPIANGIQIRLSATDGRMVAGFGGLETCGRIWLCPVCSA
                     KVRVRRGDQIAEACARHLDNEGAACWFITVTLPHEKGDALKDSFTALNEAWRYVKTGKA
                     YQADKKRFGILGDIRAVEVTHGENGWHPHAHILVFTTKQLEFDEMCAWAARLDARWARG
                     LAKVGWPQGKPGVRFTMVPVTKGKGVAAYVAKVQEKGLGNEIARADMKDATSGNRTPFG
                     ILADFGNDALADDLELWWEYEAATAGRSAIRWSPGLWAKLMPDEDELTDEEIAAEDEKG
                     STIAYMAPWTWRRLRTIQGAEIAVLEAVEAHGWGGLLRVLIDLRIGTQGVFTPDEWATP
                     GTVDLVG"
     gene            complement(431..1474)
                     /gene="rep"
                     /label=rep
                     /note="from pYS1"
     CDS             2434..3228
                     /codon_start=1
                     /product="aminoglycoside 3-O-phosphotransferase"
                     /function="inactivates kanamycin"
                     /label=aminoglycoside 3-O-phosphotransferase
                     /note="from pK18"
                     /protein_id="ADR00352.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      3512..4100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4388..4409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4424..4454
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4462..4478
                     /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     4486..4502
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4498..526
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRDKLADGADRYRGCEPVRVEAGPRDLG
                     GGLAGSAIAAGSRRAGRAFGRPFSCAGWWGRASPGLDCPALPMESRDASLFALDTLSSA
                     LDLACIGEGSTGVTDGGRGVYPGGSSERGRGSSGGSHLMLPGNGPISATPLAPSVNPES
                     ADEIHSSRGGPFVGREDSLSTDPEVDEHPE"
     gene            4498..16
                     /gene="lacZ alpha"
                     /label=lacZ alpha
                     /note="contains alpha complementation region; function in 
                     vector not yet confirmed"
     misc_feature    4512..4568
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4572..4588)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"