LOCUS       Exported               11849 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSTA, complete sequence.
ACCESSION   KC710297
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11849)
  AUTHORS   Baka E, Varga S, Hobel C, Knispel RW, Fekete C, Ivanics M, Kriszt B,
            Nagy I, Kukolya J.
  TITLE     The first transformation method for the thermo-acidophilic archaeon 
            Thermoplasma acidophilum
  JOURNAL   J. Microbiol. Methods 95 (2), 145-148 (2013)
  PUBMED    23978635
REFERENCE   2  (bases 1 to 11849)
  AUTHORS   Baka E, Kukolya J, Nagy I, Varga S, Knispel R.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-FEB-2013) Department of Microbiology, Central 
            Environmental and Food Science Research Institute, Herman Otto Road 
            15, Budapest 1022, Hungary
REFERENCE   3  (bases 1 to 11849)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Ape Plasmid Editor v. 2.0
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11849
                     /organism="Cloning vector pSTA"
                     /mol_type="other DNA"
                     /label=derived from Thermoplasma acidophilum
                     /note="derived from Thermoplasma acidophilum"
                     /db_xref="taxon:1328815"
     misc_feature    1..6100
                     /label=pTA1 plasmid NcoI fragment
                     /note="pTA1 plasmid NcoI fragment"
     misc_feature    1..163
                     /note="derived from pTA1 plasmid ORF13 from T. acidophilum 
                     of INSD accession AB190359"
     CDS             complement(122..457)
                     /codon_start=1
                     /product="hypothetical membrane protein"
                     /label=hypothetical membrane protein
                     /note="derived from pTA1 plasmid ORF14 from T. acidophilum 
                     of INSD accession AB190359"
                     /protein_id="AGL45415.1"
                     /translation="MCISMICLCRPVLVSCTCCFPGVCVRILGISLAPCLRSYLGRPLQ
                     YPFSYLSYYYVLVVKNIGFVVCRFWRPALVADPALRFRLLPFFITLDNVRSAPLFVRGH
                     IFFGCLI"
     CDS             500..808
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="derived from pTA1 plasmid ORF15 from T. acidophilum 
                     of INSD accession AB190359"
                     /protein_id="AGL45416.1"
                     /translation="MNSETYILIFIDNLPRKIPTWIAESKNIKEFEHFSLKKGTEKAML
                     ISNQRLEKWIPVSKTKRIKKENETALEWAIKELNNPEHRNEATAKIRKLINSEEGEQ"
     CDS             805..1044
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="derived from pTA1 plasmid ORF16 from T. acidophilum 
                     of INSD accession AB190359"
                     /protein_id="AGL45417.1"
                     /translation="MTDLKQLLKDHHPEIKEMPSHKEAIEILLQDLENIYKRMRILYYC
                     RNCKKIINPGEEYLHFQHILEDAIILQEPPPKRN"
     CDS             1004..1381
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="derived from pTA1 plasmid ORF17 from T. acidophilum 
                     of INSD accession AB190359"
                     /protein_id="AGL45418.1"
                     /translation="MPLYYKNPHQKEIDGPLIFSPTIITKSNKKNTFNKSMPYQGMKQT
                     INPEEILHLARIGEIGTIDSYKGQPLIVNPLNDRPISIQYDPHTDSFFATIGSRNIYLT
                     DKQASRLIRDLTKYLDQITKI"
     CDS             1702..2115
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="derived from pTA1 plasmid ORF18 from T. acidophilum 
                     of INSD accession AB190359"
                     /protein_id="AGL45419.1"
                     /translation="MPVINANISDEAYTILKEITEQTDGNKSKALTKIILAFKECMENH
                     QQTASEEHEEDQGSTEDIEQIKKFLQDQTEEAPERYQITKGELYQKFLEYCKKSSHPEI
                     PKPTFYSIIKRNIKIQDNDNARPRVYKGIKWKE"
     CDS             2553..3638
                     /codon_start=1
                     /gene="Cdc6"
                     /product="cell division control protein 6-like protein"
                     /label=Cdc6
                     /note="derived from pTA1 plasmid ORF1 from T. acidophilum 
                     of INSD accession AB190359; putative replication initiator"
                     /protein_id="AGL45420.1"
                     /translation="MISDKNNLDELTRKTIETLKDNGNNLKDPSVFDLNYQPKKIFIRK
                     EINAILEQLGQYALWKIPKHIALIGPKGAGKTITIKTIAESLQRTQGLENIYINARETQ
                     TSYKIYQEITKHYTKGPDLSEMRAKALKRLTDHTLLIIDEVDFLKDQDILYTVTRETKT
                     TLILLTQKLSWIKNLKDESVASSLQPQLITFNQYTPQELSEILTMRAEEGLYQFDPGSI
                     NLISALVARNYRGDTRIAIKTLERIGYRNQWNEEEIYKALEEAYNELEGTILRGLSPRD
                     LEILLIISKTPESNKAYLIAKQSTYARNISRPTFLQALNHLQNLDLIMLTKKRVGKSNT
                     QEAEILLNNKDLIEKIASEKL"
     gene            2553..3638
                     /gene="Cdc6"
                     /label=Cdc6
     CDS             complement(4196..5080)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="derived from pTA1 plasmid ORF2 from T. acidophilum 
                     of INSD accession AB190359"
                     /protein_id="AGL45421.1"
                     /translation="MRSGPCGVLPRIFRISMRAWLIKFSSIFSFSSIHDGDLRRFAMIN
                     DFLAWAVANLKPYQFIDVRNAYSERLDVPIFTLDYAGFSAYMDAVVGMNWDVRYEDEVM
                     AVLKNGPASVSDIVDRITSKYPVSRKEASDKVWLVLGRFRDNDMGWADCQGHVLKLRHI
                     DMDLKHDEGGLWFLSGSNMSVLHDWMEGQVVRVLRDYHVEFAVPPEYTRRRDADIVLHG
                     FKIEIETGLKKDSSALVDRISSYDGPVVVVVPNQAVKSRYARILSGFQNVNVLTLLEFI
                     YDLKREYIKSVFK"
     CDS             5001..5603
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="derived from pTA1 plasmid ORF3 from T. acidophilum 
                     of INSD accession AB190359"
                     /protein_id="AGL45422.1"
                     /translation="MLENLISQALIDIRKILGSTPQGPERTDLTHHIVNHCGTHTGQPV
                     DLIDGQDNRTAPRVYHPPDLTEEKLGIEGFCTLIKAGEIEDVVIIDAKGEIDHRIGRYK
                     TQVSLYNGPGLTLDPGSIRPAGPHDIQKIIIGRRNHDRRMQERRDLRSGYTLDVNGKGN
                     IPRINEGITIKKEGIRDPISNPNNTKRKMQIRIIGLK"
     misc_feature    6101..8445
                     /label=pRSF_Duet vector
                     /note="pRSF_Duet vector"
     CDS             6112..6129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        6243..6261
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6262..6286
                     /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             6395..6439
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     terminator      6491..6538
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(6768..7583)
                     /codon_start=1
                     /product="kanamicin reistance factor"
                     /label=kanamicin reistance factor
                     /note="provides kanamicin resistance in E. coli; RSF ori"
                     /protein_id="AGL45424.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(7584..7675)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7691..8440)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     misc_feature    8446..8852
                     /label=promoter Ta1137/1138
                     /note="promoter Ta1137/1138"
     CDS             8856..10781
                     /codon_start=1
                     /gene="gyrB"
                     /product="gyrase B"
                     /label=gyrB
                     /note="Ta10555"
                     /protein_id="AGL45423.1"
                     /translation="MTEDNYDSSQIQILEGLKAVRKVPGMYIGSTDTRGLHHLVYEVVD
                     NSVDESVAGYCSRIYVVMGSDGSITVEDDGRGIPVDIHPKYNRPGLEIVLTELHSGAKF
                     DKKVYKLTGGLHGVGLHVVNALSKKLIAVVKRDGKIYYDIFEQGIPVSGLKTASDVSEI
                     EKLGIKIQFPEHGAIIKFYPDPDIFETTEFSYETILARLTDLSYLNPQLTITFLDEASG
                     RKDILHHEGGLIELVRHLSEGKEALMEPLYLKEEVDSHMVEFSLLYTTDVQETLMSFVN
                     NISTPEGGTHVAGFHQGLSRAIQDYARSNNKIKGVEDITGDDVKEGVIAVLHVKMQNPQ
                     FEGQTKSKLGNSSVRGIVQSVTAKFMKTFMETNPHVADVIIGRVLSAAAAREASRKAKE
                     LVRRKSALEGGGLPGKLADCSSNDPSNSELYIVEGDSAGGSAKQARNRQYQAVLPLRGK
                     ILNVEKASDMKVVENEIIHDLVVAIGTGVKEDLNPKKLRYGKIIIMTDADVDGAHIRTL
                     LLTFFFRYARSLIENGNVFFAEPPLYRIQKGQNVRYVYSDEEKEQVSREFGPNAIIQRF
                     KGLGEMNPEQLWETTMNPKTRKLVQVTIEDAEEAERIFTILMGEKVEPRRKFIEENAVY
                     AENIDL"
     gene            8856..10781
                     /gene="gyrB"
                     /label=gyrB
     misc_feature    8856..9022
                     /gene="gyrB"
                     /note="Ta1055; derived from INSD accession AL445066"
     misc_feature    9023..9993
                     /gene="gyrB"
                     /label=derived from INSD accession AB206999
                     /note="derived from INSD accession AB206999"
     misc_feature    9994..10781
                     /gene="gyrB"
                     /note="Ta1055; derived from INSD accession AL445066"
     misc_feature    10782..11803
                     /label=pRSF_Duet vector
                     /note="pRSF_Duet vector"
     promoter        complement(11639..11716)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        11762..11780
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    11781..11805
                     /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)."