LOCUS       62056_8595        6001 bp DNA     circular SYN 27-APR-2021
DEFINITION  Suicide vector pDS261, complete sequence.
ACCESSION   MW429486
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  AUTHORS   Susanti D, Frazier MC, Mukhopadhyay B.
  TITLE     A Genetic System for Methanocaldococcus jannaschii: An Evolutionary 
            Deeply Rooted Hyperthermophilic Methanarchaeon
  JOURNAL   Front Microbiol 10, 1256 (2019)
  PUBMED    31333590
REFERENCE   2  (bases 1 to 6001)
  AUTHORS   Khairunisa BH, Susanti D, Mukhopadhyay B.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JAN-2021) Biochemistry, Virginia Tech, 340 West Campus
            Drive, Blacksburg, VA 24061, USA
REFERENCE   3  (bases 1 to 6001)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Microbiol 10, 1256 (2019)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-JAN-2021) Biochemistry, Virginia Tech, 340 West Campus Drive, 
            Blacksburg, VA 24061, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             698..1000
                     /codon_start=1
                     /transl_table=11
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="part of mj_0749; upstream of fprA (mj_0748)"
                     /protein_id="QUC68721.1"
                     /translation="AYLLLALFLVLGFIYPMFFCRYVCPVGSLLSIFARFSIFKLKLDE
                     NKCVGCRLCERKCPMQIKITEKIDQMECIRCFECMSVCKKGALSFSAFTKNTKKE"
     misc_feature    1001..1212
                     /label=upstream of fprA (mj_0748)
                     /note="upstream of fprA (mj_0748)"
     regulatory      1213..1528
                     /note="Methanocaldococcus FS.406-22 S-layer promoter (Psla,
                     mfs40622_1341)"
                     /regulatory_class="promoter"
     gene            1529..2746
                     /gene="hmgA"
                     /locus_tag="mj_0705"
                     /label=hmgA
     CDS             1529..2746
                     /codon_start=1
                     /transl_table=11
                     /gene="hmgA"
                     /locus_tag="mj_0705"
                     /product="3-hydroxy-3-methylglutaryl coenzyme A reductase"
                     /label=hmgA
                     /protein_id="QUC68722.1"
                     /translation="MENYNDILEKMLNGEIKPYQLDKMFGSKIATEIRRKFIEKKVGIE
                     FKHICNYSIDEEMAMKKNIENMIGAIQIPLGFAGPLKINGEYAKGEFYIPLATTEGALV
                     ASVNRGCSIITKCGGATVRVIDDKMTRAPCLKTKSVVDAIKVRDWIRENFERIKEVAES
                     TTRHGKLIKIEPILIVGRNLYPRFVFKTGDAMGMNMVTIATEKACNFIEGELKKEGIFV
                     KTVAVSGNACVDKKPSGMNLINGRGKSIVAEVFLTEKEVNKYLKTTSQAIAEVNRLKNY
                     IGSAISNSMGFNAHYANIIGAIFLATGQDEAHIVEGSLGITMAEVEDDGLYFSVTLPDV
                     PIGTVGGGTRVETQKECLEMLGCYGDNKALKFAEIVGAAVLAGELSLLGALAAGHLGKA
                     HQELGR"
     regulatory      2747..2780
                     /note="S-layer terminator from Methanocaldococcus FS.406-22
                     (Tsla, mfs40622_1341)"
                     /regulatory_class="terminator"
     regulatory      2781..3056
                     /note="flagellin B1B2 (mj_0891-0892) operon promoter of 
                     Methanocaldococcus jannaschii (PflaB1B2)"
                     /regulatory_class="promoter"
     CDS             3066..3131
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     CDS             3147..3170
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             3207..3230
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     primer_bind     complement(3760..3776)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3813..3831)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3852..3868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3876..3892)
                     /label=lac operator
                     /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)."
     promoter        complement(3900..3930)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3945..3966)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4254..4842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5016..5873)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5874..5978)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatgca tatctgctgt tggctttatt
      721 tttagtttta ggatttattt atccaatgtt cttctgcaga tatgtttgtc cagtggggtc
      781 tttgttgagt atatttgcga gattctcaat ctttaaactg aaacttgatg aaaataaatg
      841 tgtaggttgt agattgtgtg aaagaaaatg tccaatgcag ataaaaataa cagaaaaaat
      901 agaccagatg gagtgtataa gatgttttga atgcatgagt gtatgtaaaa aaggagcatt
      961 atctttttca gcttttacta aaaatactaa aaaagaataa ttccaatata cacatatttg
     1021 aaaaataaaa ttaaaacaat ttatataaat ctttaagtaa attttattta tattgtgaaa
     1081 tagtattttc aaatttagca gaggtattta aatagatatg ttgaaattcg aatagttaag
     1141 ttcttatact acttacacaa taaaataaat aactgataat aaaataacta ataactacaa
     1201 atataattaa agaaatattc accaaaatta atatttattt ttggtctttt ttaacattat
     1261 tcctcatagg ttttataaat ttctactaaa tatttggata tatcaaaatt cgtttttaac
     1321 catttaaaaa agttgcaaat gtttgtaact tttttatttt agaatttgtc aaaaattata
     1381 tgataaattt gaaatatcaa agatataata gaaattcata tcgtaaaact attgttataa
     1441 ccgtttcata tcgtaagctt tatatagtag ttcgtcgaag gtatataccg tcaatcaaat
     1501 aacaacataa aaacttaggt gataaggtat ggaaaattat aatgacatcc ttgaaaaaat
     1561 gttaaatgga gaaataaagc catatcaatt agataaaatg tttggttcaa aaatagcaac
     1621 tgaaattaga agaaaattta ttgaaaaaaa ggttgggatt gaatttaagc atatatgcaa
     1681 ttactcaata gatgaagaaa tggctatgaa gaagaatata gaaaatatga ttggagctat
     1741 acagattcca ttgggttttg ctgggccttt aaagataaat ggggaatatg caaagggaga
     1801 gttttacatc ccattggcaa caactgaagg agctttggtg gcatcggtta ataggggttg
     1861 ctcaataata acaaaatgtg ggggggcaac tgttagggtt atagatgata agatgacaag
     1921 agccccttgc ttaaaaacaa agagtgttgt agatgcaata aaagtcagag attggattag
     1981 agaaaacttt gagaggataa aggaagttgc tgaatcaaca acaaggcatg gaaagctaat
     2041 aaagatagag ccaattttaa tcgttggaag aaatctatat ccaagatttg tatttaaaac
     2101 tggagatgcc atgggcatga atatggttac aattgcaaca gagaaggcat gtaattttat
     2161 agagggggaa ttaaaaaaag aaggcatatt tgttaaaaca gttgctgtca gtggaaatgc
     2221 atgtgtagat aaaaagccta gtggaatgaa cttaattaat ggtagaggga aatctattgt
     2281 agcagaggta tttttaactg agaaggaagt taataaatac ctaaaaacca cttcccaagc
     2341 tatagctgaa gtaaataggt taaagaatta tataggttca gcaataagca attcaatggg
     2401 attcaatgcc cattatgcaa atatcattgg ggcaatattc ttagctactg ggcaagatga
     2461 ggcacatata gttgagggta gtttaggaat tacaatggct gaagttgaag atgatggatt
     2521 atatttttca gtaactctcc cagatgttcc tattgggact gttggaggag gaacaagggt
     2581 tgaaacacaa aaagagtgct tagagatgct tggttgctac ggagataata aagctttaaa
     2641 atttgctgaa attgttggag ctgcagtttt agctggggag ttatctttat taggagcttt
     2701 agcagctgga catttaggaa aagctcatca agaactcggt agataaatcc tctaaatctt
     2761 tttttatttt tttagatgac gtttatccat tagtttatat aaatttggat ttgtttaaat
     2821 ttatttggca taatcattac gttaatttgc tatttaagat tcgactatcg ttttacctac
     2881 acaaacctct acataaacac cgataaaatt gactacgcaa taaattacgc aaaaagaata
     2941 tattttaaga aaattacagt ttatagtgag gtggattgag gaaagtgtgg gggaattgtg
     3001 tgggggagga aaatactaca agcaattgag ggattaatat actcaggtga tatgagatga
     3061 agatggacta taaggatcac gatggagatt ataaagatca tgatattgat tataaggatg
     3121 acgatgataa aggatcagct gcttcatggt cacacccaca atttgaaaaa ggaggtggat
     3181 caggaggtgg ttctggtgga ggatcttggt ctcatccaca attcgagaag tcaggaatga
     3241 aaaaatatga aagcagacgc agcaaaatag ctgatggtgt atattgggtg ggggttttag
     3301 actgggacat aagaatgtat cacggctaca cattaaaagg aacaacatac aatgcctatt
     3361 tagtctttgg agatgaaaaa gttgctttaa tagacaacac atacccagga acctccgctc
     3421 aaatgtgggg gaggataaaa gatgcatttg aaaaagaggg gagggaattt aaaattgatg
     3481 taatcgttca aaaccacgta gaaaaagacc acagtggagc tctccctgaa atacacaaaa
     3541 aattcccaga tgcaccaata tactgtactg aggtagctgt tgagggactt aaaaagcact
     3601 atccatcatt aaaagacgct caatttaagg ttgttcatac aggagataca gttgatttag
     3661 gaggaaagac attaacattc ttagaagctc ctctattaca ctggccagat agtatgttta
     3721 ccttctacaa cgaagggatc gaattcctgc agcccggggg atccactagt tctagagcgg
     3781 ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     3841 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3901 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3961 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4021 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4081 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4141 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4201 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4261 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4321 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4381 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4441 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4501 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4561 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4621 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4681 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4741 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4801 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4861 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4921 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4981 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5041 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5101 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5161 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5221 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5281 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5341 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5401 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5461 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5521 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5581 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5641 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5701 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5761 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5821 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5881 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5941 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6001 c
//
