LOCUS       sequence-39504-e        6996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6996)
  AUTHORS   Kobayashi T, Antar AA, Boehme KW, Danthi P, Eby EA, Guglielmi KM, 
            Holm GH, Johnson EM, Maginnis MS, Naik S, Skelton WB, Wetzel JD, 
            Wilson GJ, Chappell JD, Dermody TS
  TITLE     A plasmid-based reverse genetics system for animal double-stranded 
            RNA viruses.
  JOURNAL   Cell Host Microbe. 2007 Apr 19;1(2):147-57.
  PUBMED    18005692
REFERENCE   2  (bases 1 to 6996)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Host 
            Microbe."; date: "2007-04-19"; volume: "1(2)"; pages: "147-57"
COMMENT     Sequence Label: pT7-L2T3D
FEATURES             Location/Qualifiers
     source          1..6996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             779..787
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(4097..4115)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4111..4158
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(4226..4244)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4227..4244)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4535..4554)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4765..4784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4884..4906
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4944..4962)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5030..5134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5135..5203,5204..5995)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5353..5372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6166..6754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6655..6674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6908..6925
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgatcccg ggaattctaa tacgactcac tatagctaaa aggcgcgatg gcgaacgttt
       61 ggggggtgag acttgcagac tcgttatctt cacccactat tgagacacga acgcgtcagt
      121 ataccttaca cgatctttgc tcagacctag atgctaatcc ggggagggaa ccgtggaaac
      181 ctctgcgtaa tcagcgtact aataatattg tggctgtgca attattcaga ccattgcagg
      241 gtttagtttt agatacccag ctttatggat ttccaggagc atttgatgac tgggagcgat
      301 tcatgagaga gaagctgcgt gtgctaaagt atgaagtatt gcgcatctat ccaatcagca
      361 actatagcaa tgaacatgtc aacgtcttcg tggccaatgc tttggtgggc gctttcctgt
      421 cgaatcaagc tttctatgac ctgctaccgt tgttgataat taatgacact atgattggtg
      481 atctacttgg cacgggggca tcgctatcac agttctttca atctcatgga gatgtgctgg
      541 aagtcgcagc tggtcgtaag tatctgcaga tggaaaacta ctccaacgat gacgatgatc
      601 ctccattatt tgcgaaagac ctgtcagatt atgctaaagc attctacagt gacacatatg
      661 aagtgttgga caggttcttt tggacgcatg actcttcagc gggggtctta gtgcattatg
      721 ataagccaac gaatggtcat cactatctgc tgggtacttt gactcagatg gtcagtgcac
      781 ctccttatat tattaacgct actgacgcaa tgttgcttga atcctgtcta gaacagttct
      841 cagctaatgt gcgtgcgaga cctgcgcaac ccgttacacg cttagaccaa tgctatcatt
      901 taagatgggg agcacaatat gtaggagaag attcactgac atatcggttg ggggtgttat
      961 ccttgctggc taccaatgga tatcaattag ctagaccgat tccaagacag ttgacgaatc
     1021 gatggttgtc gagctttgtg agtcaaatta tgtctgacgg cgtcaacgag actccactgt
     1081 ggccccaaga aaggtatgtg cagatcgctt atgattcacc atccgttgtt gatggggcta
     1141 cgcaatatgg ctatgtcagg aagaatcaac tcagactcgg catgagaata tcggcgctgc
     1201 aatcgctgag tgatacgccc tcgccggtac agtggcttcc acaatacacc atcgaccagg
     1261 cagcgatgga cgaaggcgat ctgatggtta gtcggcttac gcaactcccg ttacgtcctg
     1321 attatggtaa tatctgggtc ggcgatgcgc tatcctatta tgtggactac aatcggagtc
     1381 atcgagtcgt gctttcatcg gaacttcctc agcttccgga cacatatttt gatggcgatg
     1441 aacagtatgg gcgcagcctg ttctcactag ctcgtaagat tggtgaccgc tcgttagtga
     1501 aagatacggc tgtcttgaag cacgcttacc aagccatcga tccaaatact ggtaaggggt
     1561 atctgagatc tgggcaatct gtcgcatatt ttggtgcatc agcgggtcat tctggtgccg
     1621 accagccgtt agtcatagag ccctggattc aagggaaaat cagtggtgtg ccgccaccct
     1681 cctcagtgcg acagttcggc tatgatgttg cccgtggcgc gatcgtcgat ctggcgagac
     1741 catttccttc tggagattat caatttgtct attcggatgt tgaccaggtg gtcgatggcc
     1801 atgacgatct gagtatatca tctggactgg tggagagcct tttgtcttca tgcatgcacg
     1861 ccacagcacc cgggggctca tttgttgtta agataaattt tccgactaga cccgtatggc
     1921 actacatcga acagaagatc ttgcccaata ttacgtcata catgttgatc aagcctttcg
     1981 tcaccaacaa cgtcgaattg ttcttcgtcg ctttcggtgt gcatcaacac tcatcactta
     2041 cttggacatc tggagtgtac ttcttcttgg tggaccattt ttatcgttat gagactttat
     2101 ctacgatctc acgacaattg ccgtcttttg ggtatgttga tgatgggtct tccgtgactg
     2161 gtatcgagac aattagtatt gagaaccctg gcttctcgaa tatgacccag gccgctcgca
     2221 ttggtatctc aggattgtgt gctaatgtag gtaacgcgcg taagtccatt gccatttacg
     2281 aatcccatgg ggccagagta ttaactatca catcaaggag atctccggca tcagctagaa
     2341 gaaagtctag gttgcgatat ttgccattaa tagaccctag gtcgttagag gtacaggcgc
     2401 gcactattct gccagctgat ccagtgttat ttgaaaacgt gagcggagcg tcaccccatg
     2461 tttgtctgac aatgatgtac aacttcgaag tgtcgtcagc ggtatatgat ggagacgttg
     2521 tgctagatct tgggacggga ccagaggcta aaatccttga actgataccc gcaacctctc
     2581 cagtcacatg cgtggacata cggcctacag cgcagcctag tggatgttgg aacgttcgta
     2641 ccacgttcct tgagttagat tatttgagcg atggatggat cactggggtg cgtggggaca
     2701 tagttacttg tatgttatct ttgggggccg ctgccgctgg aaaatcaatg acttttgacg
     2761 ctgcgtttca gcaattaatc aaagtattat ccaagagtac ggctaatgtt gtgctggtgc
     2821 aggttaactg ccctacagac gtggtgagga gcattaaggg ctacctagag atagattcga
     2881 ctaacaagag gtataggttc cccaaatttg gtcgagacga gccgtactct gacatggatg
     2941 cgctggagaa aatatgtcgt accgcctggc caaactgctc aattacctgg gttccattgt
     3001 catacgactt gcggtggact agactggcat tattagagtc cacgacattg agtagcgcgt
     3061 cgattagaat tgctgagctg atgtataaat acatgcctat tatgaggatt gacattcatg
     3121 gactacccat ggaaaagcga ggtaacttca tagtggggca gaactgctca ttagtaatcc
     3181 ctggttttaa tgcgcaggat gtctttaact gttatttcaa ttccgccctc gctttctcga
     3241 ctgaagatgt caatgctgcg atgattcccc aagtgtctgc gcagtttgat gcgactaagg
     3301 gtgagtggac gttggatatg gtcttctccg acgcaggaat ctataccatg caggctctag
     3361 tgggatctaa tgctaatcca gtctctttgg gttcctttgt agttgattct ccagatgtag
     3421 atataactga cgcttggcca gctcagttag actttacgat cgcgggaact gatgtcgata
     3481 taacagttaa tccttattac cgtctgatga cctttgtaag gatcgatgga cagtggcaga
     3541 ttgccaatcc agacaaattt caattctttt cgtcggcgtc tgggacgtta gtgatgaacg
     3601 tcaaattaga tatcgcagat aaatatctac tatactatat acgagatgtc cagtctcgag
     3661 atgttggctt ttacattcag catccacttc aacttttgaa tacgatcaca ttgccaacca
     3721 acgaggacct ttttctgagc gcacctgaca tgcgagagtg ggcagttaag gaaagcggta
     3781 acacgatatg tatactcaat agtcaagggt ttgtgctacc tcaagattgg gatgtgttaa
     3841 cagataccat aagttggtcc ccatcgatac ccacatacat tgtgccaccg ggtgattata
     3901 ccttgactcc tctgtaactc actgtccctc gtgagcgcgc ctaattcatc gggtcggcat
     3961 ggcatctcca cctcctcgcg gtccgacctg ggcatccgaa ggaggacgtc gtccactcgg
     4021 atggctaagg gagagctcaa aaaaaaggat ccggctgcta acaaagcccg aaaggaagct
     4081 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     4141 gtcttgaggg gttttttgct gaaaggagga actatatccg gatcgagatc ctctagagtc
     4201 gacctgcagg catgcaagct tgtattctat agtgtcacct aaatcgtatg tgtatgatac
     4261 ataaggttat gtattaattg tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg
     4321 tagcatctgg cttactgaag cagaccctat catctctctc gtaaactgcc gtcagagtcg
     4381 gtttggttgg acgaaccttc tgagtttctg gtaacgccgt tccgcacccc ggaaatggtc
     4441 agcgaaccaa tcagcagggt catcgctagc cagatcctct acgccggacg catcgtggcc
     4501 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     4561 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     4621 ggccccgtgg ccgggggact gttgggcgcc atctccttgc accattcctt gcggcggcgg
     4681 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
     4741 agcgtcgata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4801 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4861 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4921 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4981 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5041 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5101 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5161 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5221 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5281 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5341 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5401 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5461 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5521 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5581 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5641 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5701 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5761 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5821 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5881 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5941 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6001 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6061 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6121 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6181 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6241 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6301 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6361 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6421 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6481 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6541 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6601 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6661 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6721 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6781 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6841 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6901 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6961 ccccgcgcgt tggccgattc attaatgcag ggggat
//
