LOCUS       62056_8590        5439 bp DNA     circular SYN 27-APR-2021
DEFINITION  Suicide vector pDS210, complete sequence.
ACCESSION   MW413896
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5439)
  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 5439)
  AUTHORS   Khairunisa BH, Susanti D, Mukhopadhyay B.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-DEC-2020) Biochemistry, Virginia Tech, 340 West Campus
            Drive, Blacksburg, VA 24061, USA
REFERENCE   3  (bases 1 to 5439)
  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 
            (26-DEC-2020) 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..5439
                     /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"
     misc_feature    665..666
                     /note="part of radA; downstream of fsr (mj_0870)"
     gene            667..1098
                     /gene="radA"
                     /locus_tag="mj_0869"
                     /label=radA
     CDS             667..1098
                     /codon_start=1
                     /transl_table=11
                     /gene="radA"
                     /locus_tag="mj_0869"
                     /product="RadA"
                     /label=radA
                     /note="downstream of fsr (mj_0870)"
                     /protein_id="QUC68720.1"
                     /translation="ARAYNSDMQMLYAENVENLIREGHNIKLVIVDSLTSTFRTEYIGR
                     GKLAERQQKLGRHMATLNKLADIYNCVVIVTNQVAARPDALFGPSEQAIGGHIVGHAAT
                     FRIFLRKAKGDKRVAKLYDSPHLPDAEAMFRITEKGIHD"
     misc_feature    1099..1164
                     /label=downstream of fsr (mj_0870)
                     /note="downstream of fsr (mj_0870)"
     misc_feature    1165..1172
                     /label=AscI restriction site
                     /note="AscI restriction site"
     gene            complement(1173..2390)
                     /gene="hmgA"
                     /locus_tag="mj_0705"
                     /label=hmgA
     CDS             complement(1173..2390)
                     /codon_start=1
                     /transl_table=11
                     /gene="hmgA"
                     /locus_tag="mj_0705"
                     /product="3-hydroxy-3-methylglutaryl coenzyme A reductase"
                     /label=hmgA
                     /protein_id="QUC68718.1"
                     /translation="MENYNDILEKMLNGEIKPYQLDKMFGSKIATEIRRKFIEKKVGIE
                     FKHICNYSIDEEMAMKKNIENMIGAIQIPLGFAGPLKINGEYAKGEFYIPLATTEGALV
                     ASVNRGCSIITKCGGATVRVIDDKMTRAPCLKTKSVVDAIKVRDWIRENFERIKEVAES
                     TTRHGKLIKIEPILIVGRNLYPRFVFKTGDAMGMNMVTIATEKACNFIEGELKKEGIFV
                     KTVAVSGNACVDKKPSGMNLINGRGKSIVAEVFLTEKEVNKYLKTTSQAIAEVNRLKNY
                     IGSAISNSMGFNAHYANIIGAIFLATGQDEAHIVEGSLGITMAEVEDDGLYFSVTLPDV
                     PIGTVGGGTRVETQKECLEMLGCYGDNKALKFAEIVGAAVLAGELSLLGALAAGHLGKA
                     HQELGR"
     regulatory      complement(2391..2706)
                     /note="Methanocaldococcus FS.406-22 S-layer promoter
                     (Psla)"
                     /regulatory_class="promoter"
     misc_feature    2707..2714
                     /label=AscI restriction site
                     /note="AscI restriction site"
     misc_feature    2715..2929
                     /label=upstream of fsr (mj_0870)
                     /note="upstream of fsr (mj_0870)"
     gene            2930..3214
                     /locus_tag="mj_0871"
                     /label=mj_0871
     CDS             2930..3214
                     /codon_start=1
                     /transl_table=11
                     /locus_tag="mj_0871"
                     /product="hypothetical protein"
                     /label=mj_0871
                     /note="part of mj_0871; upstream of fsr (mj_0870)"
                     /protein_id="QUC68719.1"
                     /translation="MKISAYYTIRISIIVLTTVFIVNYIMSTGIMKKLSNMLSPILRRL
                     KVNPLSISSTLACFFSPTVGYSILAEGLKENKVNEREVIGASLANSFPSV"
     promoter        complement(3251..3269)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3290..3306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3314..3330)
                     /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(3338..3368)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3383..3404)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3692..4280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4454..5311)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5312..5416)
                     /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 gccctagcaa gagcctacaa ctcagatatg caaatgttgt atgctgaaaa tgttgagaat
      721 ttaataagag aagggcataa tataaagttg gttatagttg attcattaac atcaacattc
      781 agaactgagt atataggaag aggtaaatta gcagagagac aacaaaaatt aggaaggcac
      841 atggctactc tcaacaaatt agctgatata tacaactgtg ttgttatagt aactaaccaa
      901 gtagctgcaa gaccagatgc tttatttgga ccttcagagc aggcaattgg agggcatatt
      961 gttgggcatg cagcaacatt tagaatattc ctaagaaagg caaaaggaga taagagggtt
     1021 gctaagcttt atgattctcc acacttacca gatgctgaag caatgtttag gataaccgaa
     1081 aagggaattc atgattaatt ctttaaatta atttttaaaa tatatcttta aaaaataaaa
     1141 gaaataaaac atctaaaaaa ttaaggcgcg ccttatctac cgagttcttg atgagctttt
     1201 cctaaatgtc cagctgctaa agctcctaat aaagataact ccccagctaa aactgcagct
     1261 ccaacaattt cagcaaattt taaagcttta ttatctccgt agcaaccaag catctctaag
     1321 cactcttttt gtgtttcaac ccttgttcct cctccaacag tcccaatagg aacatctggg
     1381 agagttactg aaaaatataa tccatcatct tcaacttcag ccattgtaat tcctaaacta
     1441 ccctcaacta tatgtgcctc atcttgccca gtagctaaga atattgcccc aatgatattt
     1501 gcataatggg cattgaatcc cattgaattg cttattgctg aacctatata attctttaac
     1561 ctatttactt cagctatagc ttgggaagtg gtttttaggt atttattaac ttccttctca
     1621 gttaaaaata cctctgctac aatagatttc cctctaccat taattaagtt cattccacta
     1681 ggctttttat ctacacatgc atttccactg acagcaactg ttttaacaaa tatgccttct
     1741 ttttttaatt ccccctctat aaaattacat gccttctctg ttgcaattgt aaccatattc
     1801 atgcccatgg catctccagt tttaaataca aatcttggat atagatttct tccaacgatt
     1861 aaaattggct ctatctttat tagctttcca tgccttgttg ttgattcagc aacttccttt
     1921 atcctctcaa agttttctct aatccaatct ctgactttta ttgcatctac aacactcttt
     1981 gtttttaagc aaggggctct tgtcatctta tcatctataa ccctaacagt tgccccccca
     2041 cattttgtta ttattgagca acccctatta accgatgcca ccaaagctcc ttcagttgtt
     2101 gccaatggga tgtaaaactc tccctttgca tattccccat ttatctttaa aggcccagca
     2161 aaacccaatg gaatctgtat agctccaatc atattttcta tattcttctt catagccatt
     2221 tcttcatcta ttgagtaatt gcatatatgc ttaaattcaa tcccaacctt tttttcaata
     2281 aattttcttc taatttcagt tgctattttt gaaccaaaca ttttatctaa ttgatatggc
     2341 tttatttctc catttaacat tttttcaagg atgtcattat aattttccat accttatcac
     2401 ctaagttttt atgttgttat ttgattgacg gtatatacct tcgacgaact actatataaa
     2461 gcttacgata tgaaacggtt ataacaatag ttttacgata tgaatttcta ttatatcttt
     2521 gatatttcaa atttatcata taatttttga caaattctaa aataaaaaag ttacaaacat
     2581 ttgcaacttt tttaaatggt taaaaacgaa ttttgatata tccaaatatt tagtagaaat
     2641 ttataaaacc tatgaggaat aatgttaaaa aagaccaaaa ataaatatta attttggtga
     2701 atatttggcg cgcctttttc accactaaat gtgtgattga tttattcaca ctttaatgct
     2761 atttaataaa actatataaa acttactaat gtgttcatga attacacaca tataaatttc
     2821 ccccaaatgt gtatttatat tttttgatat tttttgaatt gagaattgaa tgttatttta
     2881 ataatattag gtgatagttg tggattacat aactccatta atggagagta tgaaaatctc
     2941 tgcttattat accataagaa tatctattat tgttttaaca acggtgttta ttgttaatta
     3001 cattatgagt actggcatca tgaagaagct aagtaatatg ttatctccaa ttttaagaag
     3061 gcttaaagtt aatcctcttt caatatcctc tactttagca tgctttttca gcccaacagt
     3121 aggatactca attttagcag agggattgaa ggagaataaa gtaaatgaaa gagaggttat
     3181 aggagcttct ttagcaaatt cattcccttc agttgcggcc gccaccgcgg tggagctcca
     3241 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     3301 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3361 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3421 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3481 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3541 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3601 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3661 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3721 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3781 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3841 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3901 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3961 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4021 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4081 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4141 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4201 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4261 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4321 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4381 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4441 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4501 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4561 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4621 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4681 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4741 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4801 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4861 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4921 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4981 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5041 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5101 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5161 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5221 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5281 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5341 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5401 aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
