LOCUS       Exported                7120 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMSW105, complete sequence.
ACCESSION   KX302629
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7120)
  AUTHORS   Wecker MS, Beaton S, Chado R, Ghirardi ML.
  TITLE     Development of a Rhodobacter capsulatus self-reporting model system 
            for optimizing light-dependent, [FeFe]-hydrogenase-driven H 
            production
  JOURNAL   Biotechnol. Bioeng. (2016) In press
  PUBMED    27531314
REFERENCE   2  (bases 1 to 7120)
  AUTHORS   Wecker MS.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAY-2016) Molecular Biology, GeneBiologics, 1047 
            Cherryvale Road, Boulder, CO 80303, USA
REFERENCE   3  (bases 1 to 7120)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7120
                     /organism="Cloning vector pMSW105"
                     /mol_type="other DNA"
                     /db_xref="taxon:1892241"
     CDS             complement(261..794)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(983..1011)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1281..1302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1317..1347
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1355..1371
                     /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     1379..1395
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1416..1434
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      1460..2094
                     /regulatory_class="promoter"
                     /note="fru promoter; derived from Rhodobacter capsulatus"
     CDS             2095..3870
                     /codon_start=1
                     /gene="hydA1"
                     /product="HydA1"
                     /label=hydA1
                     /note="hydrogenase; derived from Clostridium acetobutylicum
                     strain DSM 1731; codon-optimized for Rhodobacter 
                     capsulatus"
                     /protein_id="AOF40441.1"
                     /translation="MKTIILNGNEVHTDKDITILELARENNVDIPTLCFLKDCGNFGKC
                     GVCMVEVEGKGFRAACVAKVEDGMVINTESDEVKERIKKRVSMLLDKHEFKCGQCSRRE
                     NCEFLKLVIKTKAKASKPFLPEDKDALVDNRSKAIVIDRSKCVLCGRCVAACKQHTSTC
                     SIQFIKKDGQRAVGTVDDVCLDDSTCLLCGQCVIACPVAALKEKSHIEKVQEALNDPKK
                     HVIVAMAPSVRTAMGELFKMGYGKDVTGKLYTALRMLGFDKVFDINFGADMTIMEEATE
                     LLGRVKNNGPFPMFTSCCPAWVRLAQNYHPELLDNLSSAKSPQQIFGTASKTYYPSISG
                     IAPEDVYTVTIMPCNDKKYEADIPFMETNSLRDIDASLTTRELAKMIKDAKIKFADLED
                     GEVDPAMGTYSGAGAIFGATGGVMEAAIRSAKDFAENKELENVDYTEVRGFKGIKEAEV
                     EIAGNKLNVAVINGASNFFEFMKSGKMNEKQYHFIEVMACPGGCINGGGQPHVNALDRE
                     NVDYRKLRASVLYNQDKNVLSKRKSHDNPAIIKMYDSYFGKPGEGLAHKLLHVKYTKDK
                     NVSKHENWSHPQFEK"
     gene            2095..3870
                     /gene="hydA1"
                     /label=hydA1
     CDS             3844..3867
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(3915..3933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3943..3959)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4666..5328)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      5329..6098
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"