LOCUS       Exported               10933 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMSW107, complete sequence.
ACCESSION   KX302630
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10933)
  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 10933)
  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 10933)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10933
                     /organism="Cloning vector pMSW107"
                     /mol_type="other DNA"
                     /db_xref="taxon:1892242"
     CDS             458..1252
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    1657..1678
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1693..1723
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1731..1747
                     /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     1755..1771
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1792..1810
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(1829..2770)
                     /codon_start=1
                     /gene="HupSL-emGFP"
                     /product="HupSL-emerald green fluorescent protein fusion"
                     /label=HupSL-emGFP
                     /note="contains reporter protein emerald GFP"
                     /protein_id="AOF40445.1"
                     /translation="MSDIETFYDVMRRQGITRRSFMKFCSLTAAALGLGPSFVPKIAEA
                     METKPRTPVVWVHGLECTARLESSTRPAHMVSKGEELFTGVVPILVELDGDVNGHKFSV
                     SGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAM
                     PEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNS
                     HKVYITADKQKNGIKVNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSA
                     LSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     gene            complement(1829..2770)
                     /gene="HupSL-emGFP"
                     /label=HupSL-emGFP
     CDS             complement(1829..2548)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    complement(2549..2770)
                     /gene="HupSL-emGFP"
                     /note="HupSL remnant; acts as signal sequence for the 
                     downstream emGFP"
     regulatory      complement(2771..3299)
                     /regulatory_class="promoter"
     primer_bind     3306..3322
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3356..3372)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3405..3423)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3433..3449)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3881..4491
                     /regulatory_class="promoter"
                     /note="fru promoter; derived from Rhodobacter capsulatus"
     CDS             4492..5544
                     /codon_start=1
                     /gene="hydE"
                     /product="HydE"
                     /label=hydE
                     /note="derived from Clostridium acetobutylicum strain DSM 
                     1731; codon-optimized for Rhodobacter capsulatus"
                     /protein_id="AOF40442.1"
                     /translation="MDNIIKLINKAEVTHDLTKDELVTLLKDDTHNEEIYKAADRVREK
                     YVGEEVHLRGLIEFSNICKRNCMYCGLRRDNKNIKRYRLEPDEIIHLAKSAKNYGYQTV
                     VLQSGEDDYYTVEKMKYIVSEIKKLNMAITLSIGEKTFEEYEEYRKSGADRYLIRIETT
                     DKELYEKLDPKMSHENRINCLKNLRKLGYEVGSGCLVGLPNQTIESLADDILFFKEIDA
                     DMIGVGPFIPNEDTPLGEEKGGEFFMSVKVTALIRLLLPDINIPATTAMESLYPNGRSI
                     ALTSGANVVMPNVTEGEYRKLYALYPGKICVNDTPGHCRQCISLKINKINRKVSATKGF
                     RKKSYKESIG"
     gene            4492..5544
                     /gene="hydE"
                     /label=hydE
     CDS             5558..6793
                     /codon_start=1
                     /gene="hydF"
                     /product="HydF"
                     /label=hydF
                     /note="derived from Clostridium acetobutylicum strain DSM 
                     1731; codon-optimized for Rhodobacter capsulatus"
                     /protein_id="AOF40443.1"
                     /translation="MNELNSTPKGERLHIALFGKTNVGKSSVINALTSQEIALVSNVKG
                     TTTDPVYKAMELLPLGPVMLIDTAGLDDISDLGELRRGKTLEVLSKTDVAILVFDVESG
                     ITEYDKNIYSLLLEKKIPLIGVLNKIDKKDYKLEDYTSQFKIPIVPISALNNKGINNLK
                     DELIRLAPENDDKFKIVGDLLSPGDIAVLVTPIDKAAPKGRLILPQQQTIRDILESDAI
                     AMVTKEFELRETLDSLRKKPKIVITDSQVFLKVAADTPKDILMTSFSILMARHKGDLIE
                     LARGARAIEDLKDGDKILIAEACTHHRQSDDIGKVKIPRWLRQKTGKKLEFDFSSGFSF
                     PPNIEDYALIVHCAGCMLNRRSMLHRIESSVKKQIPIVNYGVLIAYVQGILPRALKPFP
                     YADRIFNQSSRN"
     gene            5558..6793
                     /gene="hydF"
                     /label=hydF
     CDS             6807..8225
                     /codon_start=1
                     /gene="hydG"
                     /product="HydG"
                     /label=hydG
                     /note="derived from Clostridium acetobutylicum strain DSM 
                     1731; codon-optimized for Rhodobacter capsulatus"
                     /protein_id="AOF40444.1"
                     /translation="MYNVKSKVATEFISDEEIIDSLEYAKQNKSNRELIDSIIEKAKEC
                     KGLTHRDAAVLLECDLEDENEKMFKLAREIKQKFYGNRIVMFAPLYLSNYCVNGCVYCP
                     YHHKNKHIARKKLSQEDVKRETIALQDMGHKRLALEAGEDPVNNPIEYILDCIKTIYSI
                     KHKNGAIRRVNVNIAATTVENYKKLKDAGIGTYILFQETYNKKSYEELHPTGPKHDYAY
                     HTEAMDRAMEGGIDDVGIGVLFGLNMYKYDFVGLLMHAEHLEAAMGVGPHTISVPRIRP
                     ADDIDPENFSNAISDEIFEKIVAIIRIAVPYTGMIVSTRESKKTRERVLELGISQISGG
                     SSTSVGGYVESEPEEDNSSQFEVNDNRTLDEIVNWLLEMNYIPSFCTACYREGRTGDRF
                     MSLVKSGQIANCCQPNALMTLKEYLEDYASSNTQKNGEALIASEVEKIPNEKVKSIVKK
                     HLTELKEGQRDFRF"
     gene            6807..8225
                     /gene="hydG"
                     /label=hydG
     CDS             complement(8479..9141)
                     /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      9142..9911
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"