LOCUS       V016234                 8081 bp    DNA     circular SYN 14-MAR-2021
DEFINITION  Exported.
ACCESSION   V016234
VERSION     V016234
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8081)
  AUTHORS   Bernhards CB, Lux MW, Katoski SE, Goralski TDP., Liem AT, Gibbons
            HS.
  TITLE     barCoder: a tool to generate unique, orthogonal genetic tags for
            qPCR detection
  JOURNAL   BMC Bioinformatics 22 (1), 98 (2021)
   PUBMED   33648451
REFERENCE   2  (bases 1 to 8081)
  AUTHORS   Bernhards CB, Lux MW, Katoski SE, Goralski TDP., Liem AT, Gibbons
            HS.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2020) US Army CCDC Chemical Biological Center,
            8198 Blackhawk Rd, Gunpowder, MD 21010, USA
REFERENCE   3  (bases 1 to 8081)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "BMC
            Bioinformatics"; date: "2021"; volume: "22"; issue: "1"; pages: "98"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-SEP-2020) US Army CCDC Chemical Biological Center, 8198
            Blackhawk Rd, Gunpowder, MD 21010, USA"
FEATURES             Location/Qualifiers
     source          1..8081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    12..29
                     /label="I-SceI site"
                     /note="I-SceI site"
     oriT            180..288
                     /label="oriT"
                     /note="incP origin of transfer"
     promoter        complement(303..321)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(331..347)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      909..2231
                     /label="ts rep from pJRS233, pWV01"
                     /note="ts rep from pJRS233, pWV01"
     gene            complement(909..1058)
                     /gene="orfD"
                     /label="orfD"
     CDS             complement(909..1058)
                     /codon_start=1
                     /transl_table=11
                     /gene="orfD"
                     /product="OrfD"
                     /label="orfD"
                     /protein_id="QSS48602.1"
                     /translation="MTNKEKELFAENEELKKEIKDLKERIERYREMEVELSTTIDLLRG
                     GIIE"
     gene            complement(1055..1753)
                     /gene="repA"
                     /label="repA"
     CDS             complement(1055..1753)
                     /codon_start=1
                     /transl_table=11
                     /gene="repA"
                     /product="RepA"
                     /label="repA"
                     /protein_id="QSS48609.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     misc_difference 1578..1595
                     /note="mutations in repA leading to temperature
                     sensitivity"
     gene            complement(1820..1981)
                     /gene="orfC"
                     /label="orfC"
     CDS             complement(1820..1981)
                     /codon_start=1
                     /transl_table=11
                     /gene="orfC"
                     /product="OrfC"
                     /label="orfC"
                     /protein_id="QSS48603.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     gene            complement(2022..2231)
                     /gene="orfB"
                     /label="orfB"
     CDS             complement(2022..2231)
                     /codon_start=1
                     /transl_table=11
                     /gene="orfB"
                     /product="OrfB"
                     /label="orfB"
                     /protein_id="QSS48604.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     CDS             2365..2376
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     regulatory      2551..2556
                     /label="Ptrc"
                     /note="Ptrc"
                     /regulatory_class="minus_35_signal"
     regulatory      2574..2578
                     /label="Ptrc"
                     /note="Ptrc"
                     /regulatory_class="minus_10_signal"
     regulatory      2594..2603
                     /regulatory_class="ribosome_binding_site"
     CDS             2611..3390
                     /gene="ant1"
                     /label="Spectinomycin 9-adenylyltransferase"
                     /note="Spectinomycin 9-adenylyltransferase from
                     Staphylococcus aureus (strain N315). Accession#: P0A0D1"
     regulatory      3419..3424
                     /label="PrrnB"
                     /note="PrrnB"
                     /regulatory_class="minus_35_signal"
     regulatory      3442..3447
                     /label="PrrnB"
                     /note="PrrnB"
                     /regulatory_class="minus_10_signal"
     regulatory      3467..3472
                     /label="PFP2"
                     /note="PFP2"
                     /regulatory_class="minus_35_signal"
     regulatory      3490..3495
                     /label="PFP2"
                     /note="PFP2"
                     /regulatory_class="minus_10_signal"
     regulatory      3508..3517
                     /regulatory_class="ribosome_binding_site"
     CDS             3544..4236
                     /label="TurboRFP"
                     /note="red fluorescent protein from Entacmaea quadricolor"
     CDS             complement(4767..5714)
                     /label="Rep101"
                     /note="RepA protein needed for replication with the pSC101
                     origin"
     regulatory      complement(5745..5750)
                     /regulatory_class="minus_10_signal"
     rep_origin      complement(5762..5984)
                     /direction=LEFT
                     /label="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     misc_feature    complement(6095..6463)
                     /label="par"
                     /note="par"
     misc_feature    complement(6478..8073)
                     /label="Btk_IntergenicTarget1_Polylinker"
                     /note="Btk_IntergenicTarget1_Polylinker"
     misc_feature    complement(join(6484..7169,7206..8069))
                     /note="similar to Bacillus thuringiensis serovar kurstaki
                     sequence in accession NZ_CP010005.1"
     misc_feature    6484..6988
                     /note="similar to fatty acid--CoA ligase; BG08_RS29910"
     misc_feature    7170..7205
                     /label="polylinker"
                     /note="polylinker"
     misc_feature    7170..7175
                     /label="XhoI restriction site"
                     /note="XhoI restriction site"
     misc_feature    7176..7181
                     /label="SmaI restriction site"
                     /note="SmaI restriction site"
     misc_feature    7182..7187
                     /label="SphI restriction site"
                     /note="SphI restriction site"
     misc_feature    7188..7193
                     /label="NheI restriction site"
                     /note="NheI restriction site"
     misc_feature    7194..7199
                     /label="NcoI restriction site"
                     /note="NcoI restriction site"
     misc_feature    7200..7205
                     /label="SacI restriction site"
                     /note="SacI restriction site"
     CDS             complement(7573..7770)
                     /codon_start=1
                     /transl_table=11
                     /product="hypothetical protein"
                     /label="hypothetical protein"
                     /note="similar to BG08_RS29905"
                     /protein_id="QSS48608.1"
                     /translation="MNTTAKLINWKEHGDMIILECELNGKRFEISTYKQRIYNAHLLSD
                     DVYIRLDSSDNIIGINIYKK"
     misc_feature    complement(7905..8069)
                     /note="similar to hypothetical protein; BG08_RS29900"
ORIGIN
        1 gcatggtacc aattaccctg ttatccctat agtaccggat cgcgcttttc cgctgcataa
       61 ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat
      121 acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg
      181 ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat
      241 tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccgcg tacccaattc
      301 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
      361 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagggg
      421 atagactgta acattctcac gcataaaatc ccctttcatt ttctaatgta aatctattac
      481 cttattatta attcaattcg ctcataatta atcctttttc ttattacgca aaatggcccg
      541 atttaagcac accctttatt ccgttaatgc gccatgacag ccatgataat tactaatact
      601 aggagaagtt aataaatacg taaccaacat gattaacaat tattagaggt catcgttcaa
      661 aatggtatgc gttttgacac atccactata tatccgtgtc gttctgtcca ctcctgaatc
      721 ccattccaga aattctctag cgattccaga agtttctcag agtcggaaag ttgaccagac
      781 attacgaact ggcacagatg gtcataacct gaaggaagat cattttgttt attgcaattg
      841 tattgctatt aatcgcaaca tcaaaccaaa ataaaaaccc ccttcgactt tcgtcagggg
      901 gcttttattt attcaataat ccctcctctc aataaatcta ttgttgtact taattcaact
      961 tccatttctc tgtatctttc aatacgctct tttaagtcct taatttcttt ttttaattcc
     1021 tcattttcag caaataactc tttttctttg tttgtcattt tatttccccc gtttcagcat
     1081 caagaacctt tgcataactt gctctatatc cacactgata attgccctca aaccataatc
     1141 taaaggcgct agagtttgtt gaaacaatat cttttacatc attcgtattt aaaattccaa
     1201 actccgctcc cctaaggcga ataaaagcca ttaaatcttt tgtatttacc aaattatagt
     1261 catccactat atctaaaagt aaattcttca attctctttt ttggctttca tcaagtgtta
     1321 tatagcggtc aatatcaaaa tcattaatgt tcaaaatatc ttttttgtcg tatatatgtt
     1381 tattcttagc aatagcgtcc tttgattcat gagtcaaata ttcatatgaa cctttgatat
     1441 aatcaagtat ctcaacatga gcaactgaac tattccccaa ttttcgctta atcttgttcc
     1501 taacgctttc tattgttaca ggatttcgtg caatatatat aacgtgatag tgtggttttt
     1561 tatagtgctt tccattttgt ataatattac tattattcca tgtatcttta tctttttttt
     1621 cgtccatatc gtgtaaagga ctgacagcca tagatacgcc caaactctct aatttttctt
     1681 tccaatcatt aggaattgag tcaggatata ataaaaatcc aaaatttcta gctttagtat
     1741 ttttaatagc catgatataa ttaccttatc aaaaacaagt agcgaaaact cgtatccttc
     1801 taaaaacgcg agctttcgct tatttttttt gttctgattc ctttcttgca tattcttcta
     1861 tagctaacgc cgcaaccgca gattttgaaa aacctttttg tttcgccata tctgttaatt
     1921 ttttatcttg ctcttttgtc agagaaatca taactctttt tttcgattct gaaatcacca
     1981 tttaaaaaac tccaatcaaa taattttata aagttagtgt atcactttgt aatcataaaa
     2041 acaacaataa agctacttaa atatagattt ataaaaaacg ttggcgaaaa cgttggcgat
     2101 tcgttggcga ttgaaaaacc ccttaaaccc ttgagccagt tgggatagag cgtttttggc
     2161 acaaaaattg gcactcggca cttaatgggg ggtcgtagta cggaagcaaa attcgcttcc
     2221 tttcccccca tttttttcca aattccaaat ttttttcaaa aattttccag cgctaccgct
     2281 cggcaaaatt gcaagcaatt tttaaaatca aacccatgag ggaatttcat tccctcatac
     2341 tcccttgagc ctcctccaac cgaaatagaa gggcgctgcg cttattattt cattcagtca
     2401 tcggctttca taatctaaca gacaacatct tcgctgcaaa gccacgctac gctcaagggc
     2461 ttttacgcta cgataacgcc tgttttaacg attatgccga taactaaacg aaataaacgc
     2521 taaaacgtct cagaaacgcg gccgcagctg ttgacaatta atcatccggc tcgtataacg
     2581 tgtggaattg tgaagaaagg aggtgaaatc atgagcaatt tgattaacgg aaaaatacca
     2641 aatcaagcga ttcaaacatt aaaaatcgta aaagatttat ttggaagttc aatagttgga
     2701 gtatatctat ttggttcagc agtaaatggt ggtttacgca ttaacagcga tgtagatgtt
     2761 ctagtcgtcg tgaatcatag tttacctcaa ttaactcgaa aaaaactaac agaaagacta
     2821 atgactatat caggaaagat tggaaatacg gattctgtta gaccacttga agttacggtt
     2881 ataaatagga gtgaagttgt cccttggcaa tatcctccaa aaagagaatt tatatacggt
     2941 gagtggctca ggggtgaatt tgagaatgga caaattcagg aaccaagcta tgatcctgat
     3001 ttggctattg ttttagcaca agcaagaaag aatagtattt ctctatttgg tcctgattct
     3061 tcaagtatac ttgtctccgt acctttgaca gatattcgaa gagcaattaa ggattctttg
     3121 ccagaactaa ttgaggggat aaaaggtgat gagcgtaatg taattttaac cctagctcga
     3181 atgtggcaaa cagtgactac tggtgaaatt acctcgaaag atgtcgctgc agaatgggct
     3241 atacctcttt tacctaaaga gcatgtaact ttactggata tagctagaaa aggctatcgg
     3301 ggagagtgtg atgataagtg ggaaggacta tattcaaagg tgaaagcact cgttaagtat
     3361 atgaaaaatt ctatagaaac ttctctcaat tgatggcaaa acaacttgaa aaaagttgtt
     3421 gacaaaaaag aagctgaatg ttatattagt aaagtctcga tgtttgttga cagtataaag
     3481 ttagaaactt ataatgataa gaattttaga aaggaggtaa tttagatgcg tggatcagga
     3541 tctatgagtg aattgattaa agaaaatatg catatgaaat tatatatgga aggaactgtt
     3601 aataatcatc atttcaaatg tacatcagag ggagaaggaa aaccatatga gggaacgcaa
     3661 actatgaaaa taaaggtggt agaaggaggg cctttaccat ttgcattcga tattttagct
     3721 acgagtttta tgtatggaag taaagcattt attaatcata ctcaggggat accggatttc
     3781 tttaaacaat catttcctga aggatttaca tgggaacgta ttacaacata tgaagatggg
     3841 ggagtattaa cggcaactca agatacgtca tttcaaaatg gatgcatcat atacaatgtt
     3901 aaaatcaatg gggttaactt tccatctaat ggacctgtga tgcagaagaa aacacgtggg
     3961 tgggaagcga atactgagat gttatatcca gctgatggag ggttaagagg acatagtcaa
     4021 atggcattga aattagtagg agggggatac ttacattgtt cttttaaaac tacatatcgt
     4081 tctaagaaac cagctaaaaa cttaaagatg ccggggtttc attttgtaga ccatagatta
     4141 gaacgtatta aagaagcgga taaagaaaca tatgttgagc aacatgaaat ggctgtagca
     4201 aaatattgtg atttacctag taaattaggg catagataag aattcccacg ggttttgctg
     4261 cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga tccggcttcg
     4321 ccaggtttgc cggctgaaag cgctatttct tccagaattg ccatgatttt ttccccacgg
     4381 gaggcgtcac tggctcccgt gttgtcggca gctttgattc gataagcagc atcgcctgtt
     4441 tcaggctgtc tatgtgtgac tgttgagctg taacaagttg tctcaggtgt tcaatttcat
     4501 gttctagttg ctttgtttta ctggtttcac ctgttctatt aggtgttaca tgctgttcat
     4561 ctgttacatt gtcgatctgt tcatggtgaa cagctttaaa tgcaccaaaa actcgtaaaa
     4621 gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac agttttccct
     4681 ttgatatcta acggtgaaca gttgttctac ttttgtttgt tagtcttgat gcttcactga
     4741 tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat gttctctagt
     4801 gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcatg cttactttgc
     4861 atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg
     4921 tgtagtgttt ttcttagtcc gttacgtagg taggaatctg atgtaatggt tgttggtatt
     4981 ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc catttgtcta
     5041 tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc aaccaccaat
     5101 ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca ttggttaagc
     5161 cttttaaact catggtagtt attttcaagc attaacatga acttaaattc atcaaggcta
     5221 atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa ccactcataa
     5281 atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata ttttatgaat
     5341 ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc taatttttcg
     5401 cttgagaact tggcatagtt tgtccactgg aaaatctcaa agcctttaac caaaggattc
     5461 ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac accataagca
     5521 ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga taccgtccgt
     5581 tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca taaaattagc
     5641 ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca
     5701 actaattcag acatacatct caattggtct aggtgatttt aatcactata ccaattgaga
     5761 tgggctagtc aatgataatt actagtcctt ttcctttgag ttgtgggtat ctgtaaattc
     5821 tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa ttccgctaga
     5881 cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat aaagaaagaa
     5941 taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact ttagtcagtt
     6001 ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa
     6061 aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa tattcctttt
     6121 gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt tcgctgcgct
     6181 cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat ggattcatgc
     6241 aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc gttttatggc
     6301 gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc cctctgattt
     6361 tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct aatgcaccca
     6421 gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtcgggaatt gatgcagaag
     6481 cttagaacca gaggatgtat tacctcctta tgaagtggga gaaattattt tgcgcggccc
     6541 aactatgatg gctggctatc ataatcgtga agaagcgaat aaaaaatcga tgtataaagg
     6601 atggtatcat tcgggtgatc taggctattt tgataaagac ggttatttat tcgttgctga
     6661 tcgcgttgat gatatggtaa ttagcggtgg cgtaaatatt tatcctcgtg aaattgaaga
     6721 tttccttcat agtcatcctg gtatattaga tgtggcggta cttggtgagc cagacgaatt
     6781 atggggagag cgtgttgtgg cagttattgt gaagaaagat gaaacaataa cagaagctga
     6841 tttagaaacg tattgcaaag aaagtgatga actagcagat tataaacgtc cgcgtcatta
     6901 tttatttgta gatgaactcc ctcgtaatgc gagcgggaag ttgcagaaat ttgtgctgag
     6961 ggagtcattg aaaggtgcga aaaaataaaa aatgaaggat accccatatc tatggggtac
     7021 ccttcatttt tctatattaa acagaagaaa caatatcttt gttaattctc atttcttctt
     7081 gttgaaggtt aattagcgca tttgaaataa ggtggaaaaa caaaatggat gtaatgtcca
     7141 aaaagaaaag acaccttaag gtgcctctcc tcgagcccgg ggcatgcgct agcccatggg
     7201 agctctccga cttgaaccat ttcatttcat acgaatctat ccgtattacc attacccctt
     7261 tgttatgagt gtaaacacat actataaagc agaatcgtct atatacacta gattgttagt
     7321 tcacaaaaga caagagattt gagatcaata ttgaagtgac gccacatccc ctataataat
     7381 tgatatgtta cgccttttaa aagaatctgc tttatagaac gtgcccacaa gttatatttt
     7441 actacatgta atattcattt agtgtatttt ttgatgaata tgtataatta cataaactgt
     7501 atgaaaaaaa gagagtatac tctgatgttg tgtaataaag ataaagcccg cctacggaat
     7561 agggggcttt tcttactttt tataaatatt tattccaatt atattatcag atgagtctaa
     7621 acgaatataa acgtcatctg ataaaaggtg tgcattatat attctttgtt tatatgttga
     7681 tatctcgaag cgttttccat ttaattcaca ttctagaata atcatatcgc catgttcttt
     7741 ccaatttatt aatttagcag tcgtattcat ataaacctcc tctgaatcat tttatgaaaa
     7801 ttagtagaga tagtgtgaga aaggagttta tataacaaaa aaagacacct taagtgtctt
     7861 ctttgattac tataaaagtc gttgaatgaa ataggtttct atttttattt gcgtgaaaag
     7921 ccttcttcat caaggtattc cgcagcttct ggataagagt tgaatgtacc gtctaagttt
     7981 acaccagcgt acacgttcca catgtcatcc tctgaaataa gagtaagaat atgaccatct
     8041 tcgttctgcc attcttcttc tttcgcagga tccgtcgaca t
//
