LOCUS       sequence-39514-e        6990 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 6990)
  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 6990)
  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-L2T1L
FEATURES             Location/Qualifiers
     source          1..6990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4091..4109)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4105..4152
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(4220..4238)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4221..4238)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4529..4548)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4759..4778)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4878..4900
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4938..4956)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5024..5128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5129..5197,5198..5989)
                     /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(5347..5366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6160..6748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6649..6668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6902..6919
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgatcccg ggtaatacga ctcactatag ctattggcgc aatggcgaac gtttggggag
       61 tgagacttgc agactcttta tcatcaccca ctattgagac aagaactcgt cattacacac
      121 tacgcgattt ctgttccgac ctggatgctg tggctggcaa ggaaccctgg agacctttgc
      181 gcaatcagag aacgaatgac attgtcgccg ttcaattgtt taggccactg cagggattgg
      241 tgcttgacac gcagttctat ggattccctg gcattttctc agaatgggag cagtttataa
      301 aggaaaaact ccgcgtatta aaatatgagg ttttgcggat ctacccgatc agtaattata
      361 atcatgaacg tgtcaatgtc ttcgtagcaa atgctcttgt cggtgcgttc ctatccaacc
      421 aagccttcta tgacctgttg cctctattag taataaatga taccatgata aatgacttac
      481 ttgggacagg tgccgccctt tctcaatttt ttcaatccca tggtgaggtt ttagaagttg
      541 ccgcaggaag gaagtacctg caaatgaaga actactcgaa cgatgacgat gatccacctt
      601 tgttcgctaa agatctgtcg gactatgcga aggcgtttta cagtgacacg tttgagactc
      661 tagaccgatt tttctggaca catgactcat ctgcgggcgt cctagtgcac tatgataagc
      721 ccactaacgg gaatcattac atcttgggta ctctgacgca gatggttagt gcgcctccgc
      781 atatcattaa cgctactgac gcattgttgc tcgaatcgtg cttagaacaa tttgcggcga
      841 atgtgagagc caggccagcg cagcctgttg cgagattgga tcaatgttat catttacggt
      901 ggggtgctca atatgttggc gaggactcat tgacgtaccg tttgggggta ctttcgctgc
      961 tggctaccaa cggatatcaa ttagctagac cgatccctaa gcaattgacg aatcgatggc
     1021 tctctagttt tgtcagtcag gtaatgtcgg atggtgtaaa tgagacgcca ttatggcctc
     1081 aagagagata tgtccaaata gcctacgatt caccatctgt agtcgatgga gccacgcact
     1141 atggctatgt taggcgaaat cagttgcggt tgggcatgag ggtgtccgct cttcagtcat
     1201 tgagtgatac tccggctccg atacagtggt taccgcagta tactattgaa caggcagctg
     1261 ttgatgaggg agatctaatg gtttctcgct tgactcaact accattacgt cctgactatg
     1321 gcagcatatg ggttggtgat gccctatcgt attatgttga ttacaatcgc agccatagag
     1381 ttgtactatc atccgaacta ccacagctac cagatacata ctttgacgga gatgagcaat
     1441 acggtcgtag cttgttctct ctagcacgaa aaattggtga tcgatctctc atcaaagata
     1501 cagcagtgct taagcatgcg taccaggcca ttgatccaaa cactggaaag gaataccttc
     1561 gcgcaggaca gtccgttgca tactttgggg catcagctgg tcattcaggg gcggatcagc
     1621 ctctagtaat tgagccatgg acgcagggta agattagtgg tgtgcctccg ccctcctcag
     1681 ttagacagtt tgggtatgat gttgctaaag gtgcgatcgt agacttagca agaccgttcc
     1741 cgtcgggcga ctaccaattt gtatattctg acgtcgatca ggtcgttgat ggccacgatg
     1801 atctcagcat atcctcagga ctggtggaga gtctattaga ctcatgcatg catgccacat
     1861 ccccaggtgg gtcattcgtg atgaagatca acttcccgac acgcactgtc tggcactata
     1921 tagagcagaa gattctccca aatattacct cgtacatgtt gattaaacca ttcgtgacta
     1981 ataatgtaga gttattcttt gtggcttttg gtgtgcatca acaatcagca ttgacatgga
     2041 cgtctggggt gtatttcttc ctggtcgatc acttctatcg atacgagaca ttgtctacga
     2101 tttcacgtca gttgccatcg ttcggatacg ttgatgatgg gtcgtctgtg acaggcattg
     2161 agatgatcag tcttgaaaat ccaggctttt caaacatgac ccaagctgca cgtgttggga
     2221 tatcagggtt gtgtgcgaat gtcggtaatg cgcgtaaatt aatatctatt catgaatccc
     2281 atggggcacg cgtgctcacc atcacatcaa gaagatctcc agcttcggcc aggcggaaag
     2341 ctcgcttacg ctatttgcca ctcgtagacc cacgatcttt ggaagtgcag gcacgtacga
     2401 tactgccatc taacccagtg ttgtttgata acgtaaacgg agcatcgcct cacgtatgtt
     2461 tgacgatgat gtataacttt gaagtgtcta gtgcggtgta tgatggtgat gtagtacttg
     2521 atcttggtac cggtcctgaa gcgaagattc tggagctgat tcctccaaca tccccggtaa
     2581 catgcgtaga cattaggcca acggcacagc cgagtggctg ttggaacgta cgtacgactt
     2641 ttctagagct tgattactta agtgacggtt ggataacggg tatacgtggc gacatcgtca
     2701 cttgcatgct gtctctgggt gctgctgctg ctggaaaatc catgacgttc gacgcggcat
     2761 tccaacagct agtgaaagtt cttactaaaa gtacagctaa cgtgttgttg attcaagtca
     2821 actgcccaac ggatgtgatc cgaacaatta agggatattt ggagatagat caaactaata
     2881 agcggtacag atttccaaag tttggccgcg atgaaccgta ctctgacatg gattcattag
     2941 agcgcatatg tcgggctgca tggccaaatt gttccatcac gtgggtgccc ttgtcctatg
     3001 acctgcgttg gactaagctt gctctacttg aatcgactac actgagcagt gcatcagtga
     3061 gaattgctga attgatgtac aagtacatgc ccgttatgag gatagatatt catgggttac
     3121 ccatggaaaa gcaaggcaat ttcatagtgg gtcagaactg ctctttgact ataccgggct
     3181 tcaacgcaca ggacgtattc aattgttact tcaactccgc gctcgccttc tctactgagg
     3241 atgttaattc tgcaatgata ccacaggtga cggctcagtt tgatgctaac aaaggagagt
     3301 ggtcattgga catggtgttc tcagacgctg gtatctacac aatgcaggca ttggtcggtt
     3361 ccaacgcaaa tcctgtgtct ctgggttcgt ttgtagtgga ctctccggat gtggacataa
     3421 cagatgcatg gcctgctcag ctggacttta ccatagctgg cactgacgtc gacattacag
     3481 ttaatcctta ttaccgcttg atggcctttg tgaggattga tggacaatgg cagattgcga
     3541 atcctgataa attccaattt ttctcatcaa gtacaggaac gttagtgatg aatgtaaagt
     3601 tagatatagc tgataggtac ttgttatact acattcgtga cgttcaatct agggatgtgg
     3661 gattttacat acagcaccca ttacagttat taaatactat tacgctgcct acaaatgagg
     3721 acttattctt gagcgctcct gacatgcgcg agtgggcagt aaaggagagt ggtaatacca
     3781 tatgcatact taatagtcaa ggttttgtgc cacctcagga ttgggatgtt cttaccgata
     3841 ctattagctg gtctccttcg ctcccaactt atgtggtgcc tccgggtgat tatactctga
     3901 cacctctgta actcattacc cctcgtgagc gtgcctaatt catcgggtcg gcatggcatc
     3961 tccacctcct cgcggtccga cctgggcatc cgaaggagga cgtcgtccac tcggatggct
     4021 aagggagagc tcaaaaaaaa ggatccggct gctaacaaag cccgaaagga agctgagttg
     4081 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4141 aggggttttt tgctgaaagg aggaactata tccggatcga gatcctctag agtcgacctg
     4201 caggcatgca agcttgtatt ctatagtgtc acctaaatcg tatgtgtatg atacataagg
     4261 ttatgtatta attgtagccg cgttctaacg acaatatgta caagcctaat tgtgtagcat
     4321 ctggcttact gaagcagacc ctatcatctc tctcgtaaac tgccgtcaga gtcggtttgg
     4381 ttggacgaac cttctgagtt tctggtaacg ccgttccgca ccccggaaat ggtcagcgaa
     4441 ccaatcagca gggtcatcgc tagccagatc ctctacgccg gacgcatcgt ggccggcatc
     4501 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
     4561 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
     4621 gtggccgggg gactgttggg cgccatctcc ttgcaccatt ccttgcggcg gcggtgctca
     4681 acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc
     4741 gatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4801 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4861 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4921 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4981 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5041 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5101 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5161 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5221 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5281 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5341 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5401 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5461 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5521 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5581 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5641 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5701 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5761 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5821 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5881 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5941 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6001 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6061 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6121 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6181 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6241 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6301 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6361 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6421 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6481 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6541 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6601 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6661 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6721 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6781 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6841 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6901 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6961 gcgttggccg attcattaat gcagggggat
//
