LOCUS       Exported                8294 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSPOM DNA, complete sequence.
ACCESSION   AB861437
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8294)
  AUTHORS   Kawamura-Nagaya K, Ishibashi K, Huang YP, Miyashita S, Ishikawa M.
  TITLE     Replication protein of tobacco mosaic virus cotranslationally binds 
            the 5 untranslated region of genomic RNA to enable viral 
            replication
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 111 (16), E1620-E1628 (2014)
  PUBMED    24711385
REFERENCE   2  (bases 1 to 8294)
  AUTHORS   Ishikawa M, Kawamura K, Tsuda S, Meshi T.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-OCT-2013) Contact:Masayuki Ishikawa National Institute
            of Agrobiological Sciences, Department of Plant Sciences, 
            Plant-Microbe Interactions Research Unit; 2-1-2 Kan-non-dai, 
            Tsukuba, Ibaraki 305-8602, Japan
REFERENCE   3  (bases 1 to 8294)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8294
                     /organism="Expression vector pSPOM"
                     /focus
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /note="This sequence represents a circular E. coli plasmid 
                     pSPOM. Infectious Tobacco mosaic virus RNA can be 
                     synthesized by in vitro transcription of AgeI-linearlized 
                     pSPOM DNA by SP6 RNA polymerase. The nucleotide 1..6395 
                     region represents the sequence of Tobacco mosaic virus 
                     strain OM RNA that was predominantly detected in direct 
                     sequencing of a population of the virus. The nucleotide 
                     6396-6419 region represents a 3 linker sequence. The 
                     nucleotide 8263-8294 region represents a 5 linker and SP6 
                     promoter sequences."
                     /db_xref="taxon:1411989"
     source          1..6395
                     /organism="Tobacco mosaic virus strain OM"
                     /strain="OM"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:12251"
     source          6420..8262
                     /organism="Cloning vector pBR322"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:47470"
     misc_feature    12..67
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             69..4919
                     /codon_start=1
                     /transl_except="(pos: 3417 .. 3419,  aa: Tyr)"
                     /product="replication protein"
                     /label=replication protein
                     /note="183-kDa (180K) protein; read through at 3417-3419"
                     /protein_id="BAO73175.1"
                     /translation="MAYTQTATTSALLDTVRGNNSLVNDLAKRRLYDTAVEEFNARDRR
                     PKVNFSKVISEEQTLIATRAYPEFQITFYNTQNAVHSLAGGLRSLELEYLMMQIPYGSL
                     TYDIGGNFASHLFKGRAYVHCCMPNLDVRDIMRHEGQKDSIELYLSRLERGGKTVPNFQ
                     KEAFDRYAEIPEDAVCHNTFQTCEHQPMQQSGRVYAIALHSIYDIPADEFGAALLRKNV
                     HTCYAAFHFSENLLLEDSYVNLDEINACFSRDGDKLTFSFASESTLNYCHSYSNILKYV
                     CKTYFPASNREVYMKEFLVTRVNTWFCKFSRIDTFLLYKGVAHKSVDSEQFYTAMEDAW
                     HYKKTLAMCNSERILLEDSSSVNYWFPKMRDMVIVPLFDISLETSKRTRKEVLVSKDFV
                     YTVLNHIRTYQAKALTYANVLSFVESIRSRVIINGVTARSEWDVDKSLLQSLSMTFYLH
                     TKLAVLKDDLLISKFSLGSKTVCQHVWDEISLAFGNAFPSVKERLLNRKLIRVAGDALE
                     IRVPDLYVTFHDRLVTEYKASVDMPALDIRKKMEETEVMYNALSELSVLRESDKFDVDV
                     FSQMCQSLDVDPMTAAKVIVAVMSNESGLTLTFERPTEANVALALQDQEKASEGALVVT
                     SREVEEPSMKGSMARGELQLAGLAGDHPESSYSRNEEIESLEQFHMATADSLIRKQMSS
                     IVYTGPIKVQQMKNFIDSLVASLSAAVSNLVKILKDTAAIDLETRQKFGVLDVASRKWL
                     IKPTAKSHAWGVVETHARKYHVALLEYDEQGVVTCDDWRRVAVSSESVVYSDMAKLRTL
                     RRLLRNGEPHVSSAKVVLVDGVPGCGKTKEILSRVNFDEDLILVPGKQAAEMIRRRANS
                     SGIIVATKDNVKTVDSFMMNFGKSTRCQFKRLFIDEGLMLHTGCVNFLVAMSLCEIAYV
                     YGDTQQIPYINRVSGFPYPAHFAKLEVDEVETRRTTLRCPADVTHYLNRRYEGFVMSTS
                     SVKKSVSQEMVGGAAVINPISKPLHGKILTFTQSDKEALLSRGYSDVHTVHEVQGETYS
                     DVSLVRLTPTPVSIIAGDSPHVLVALSRHTCSLKYYTVVMDPLVSIIRDLEKLSSYLLD
                     MYKVDAGTQ*QLQIDSVFKGSNLFVAAPKTGDISDMQFYYDKCLPGNSTMMNNFDAVTM
                     RLTDISLNVKDCILDMSKSVAAPKDQIKPLIPMVRTAAEMPRQTGLLENLVAMIKRNFN
                     APELSGIIDIENTASLVVDKFFDSYLLKEKRKPNKNVSLFSRESLNRWLEKQEQVTIGQ
                     LADFDFVDLPAVDQYRHMIKAQPKQKLDTSIQTEYPALQTIVYHSKKINAIFGPLFSEL
                     TRQLLDSVDSSRFLFFTRKTPAQIEDFFGDLDSHVPMDILELDISKYDKSQNEFHCAVE
                     YEIWRRLGFEDFLGEVWKQGHRKTTLKDYTAGIKTCIWYQRKSGDVTTFIGNTVIIAAC
                     LASMLPMEKIIKGAFCGDDSLLYFPKGCEFPDVQHSANLMWNFEAKLFKKQYGYFCGRY
                     VIHHDRGCIVYYDPLKLISKLGAKHIKDWEHLEEFRRSLCDVAVSLNNCAYYTQLDDAV
                     WEVHKTAPPGSFVYKSLVKYLSDKVLFRSLFIDGSSC"
     CDS             69..3419
                     /codon_start=1
                     /product="replication protein"
                     /label=replication protein
                     /note="126-kDa (130K) protein"
                     /protein_id="BAO73176.1"
                     /translation="MAYTQTATTSALLDTVRGNNSLVNDLAKRRLYDTAVEEFNARDRR
                     PKVNFSKVISEEQTLIATRAYPEFQITFYNTQNAVHSLAGGLRSLELEYLMMQIPYGSL
                     TYDIGGNFASHLFKGRAYVHCCMPNLDVRDIMRHEGQKDSIELYLSRLERGGKTVPNFQ
                     KEAFDRYAEIPEDAVCHNTFQTCEHQPMQQSGRVYAIALHSIYDIPADEFGAALLRKNV
                     HTCYAAFHFSENLLLEDSYVNLDEINACFSRDGDKLTFSFASESTLNYCHSYSNILKYV
                     CKTYFPASNREVYMKEFLVTRVNTWFCKFSRIDTFLLYKGVAHKSVDSEQFYTAMEDAW
                     HYKKTLAMCNSERILLEDSSSVNYWFPKMRDMVIVPLFDISLETSKRTRKEVLVSKDFV
                     YTVLNHIRTYQAKALTYANVLSFVESIRSRVIINGVTARSEWDVDKSLLQSLSMTFYLH
                     TKLAVLKDDLLISKFSLGSKTVCQHVWDEISLAFGNAFPSVKERLLNRKLIRVAGDALE
                     IRVPDLYVTFHDRLVTEYKASVDMPALDIRKKMEETEVMYNALSELSVLRESDKFDVDV
                     FSQMCQSLDVDPMTAAKVIVAVMSNESGLTLTFERPTEANVALALQDQEKASEGALVVT
                     SREVEEPSMKGSMARGELQLAGLAGDHPESSYSRNEEIESLEQFHMATADSLIRKQMSS
                     IVYTGPIKVQQMKNFIDSLVASLSAAVSNLVKILKDTAAIDLETRQKFGVLDVASRKWL
                     IKPTAKSHAWGVVETHARKYHVALLEYDEQGVVTCDDWRRVAVSSESVVYSDMAKLRTL
                     RRLLRNGEPHVSSAKVVLVDGVPGCGKTKEILSRVNFDEDLILVPGKQAAEMIRRRANS
                     SGIIVATKDNVKTVDSFMMNFGKSTRCQFKRLFIDEGLMLHTGCVNFLVAMSLCEIAYV
                     YGDTQQIPYINRVSGFPYPAHFAKLEVDEVETRRTTLRCPADVTHYLNRRYEGFVMSTS
                     SVKKSVSQEMVGGAAVINPISKPLHGKILTFTQSDKEALLSRGYSDVHTVHEVQGETYS
                     DVSLVRLTPTPVSIIAGDSPHVLVALSRHTCSLKYYTVVMDPLVSIIRDLEKLSSYLLD
                     MYKVDAGTQ"
     CDS             4903..5709
                     /codon_start=1
                     /product="movement protein"
                     /label=movement protein
                     /note="30-kDa (30K) protein"
                     /protein_id="BAO73177.1"
                     /translation="MALVVKGKVNINEFIDLTKMEKILPSMFTPVKSVMCSKVDKIMVH
                     ENESLSEVNLLKGVKLIDSGYVCLAGLVVTGEWNLPDNCRGGVSVCLVDKRMERADEAT
                     LGSYYTAAAKKRFQFKVVPNYAITTQDAMKNVWQVLVNIRNVKMSAGFCPLSLEFVSVC
                     IVYRNNIKLGLREKITSVRDGGPMELTEEVVDEFMEDVPMSIRLAKFRSRTGKKSDVRK
                     GKISSSDRSAPNKNYRNVKDFGGMSFKKNNLIDDDSETTVAESDSF"
     CDS             5712..6191
                     /codon_start=1
                     /product="coat protein"
                     /label=coat protein
                     /protein_id="BAO73178.1"
                     /translation="MSYSITTPSQFVFLSSAWADPIELINLCTNALGNQFQTQQARTVV
                     QRQFSEVWKPSPQVTVRFPDSDFKVYRYNAVLDPLVTALLGAFDTRNRIIEVENQANPT
                     TAETLDATRRVDDATVAIRSAINNLVVELIRGTGSYNRSSFESSSGLVWNSGPAT"
     misc_feature    6396..6419
                     /label=3 linker
                     /note="3 linker"
     promoter        6523..6627
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6628..7488
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7659..8247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    8263..8294
                     /label=5 linker and SP6 promoter
                     /note="5 linker and SP6 promoter"
     promoter        8278..1
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"