LOCUS       sequence-39522-e        5313 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 5313)
  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 5313)
  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-M3T1L
FEATURES             Location/Qualifiers
     source          1..5313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2125..2136)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2414..2432)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2428..2475
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2543..2561)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2544..2561)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2852..2871)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3082..3101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3201..3223
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3261..3279)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3347..3451
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3452..3520,3521..4312)
                     /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(3670..3689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4483..5071
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4972..4991
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5225..5242
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgatccct aatacgactc actatagcta aagtgaccgt ggtcatggct tcattcaagg
       61 gattctccgt caacactgtt ccagtttcca aggccaaacg tgatatatca tctcttgccg
      121 ctactcctgg aattcgctca caacccttca ctccgtctgt agacatgtct caatcgcgtg
      181 aattccttac aaaggcaatt gagcaagggt ctatgtctat accttatcag catgtgaatg
      241 ttccaaaagt tgatcgtaag gttgttagtt tggtagtgcg accgttttct tcaggtgctt
      301 tttctatctc tggagtgatt tcaccggcgc acgcttatct attggattgt ctacctcagc
      361 ttgaacaggc gatggcgttt gttgcttcac ctgaatcttt ccaggcttct gatgtcgcaa
      421 aacgctttgc tataaagccg ggcatgagcc ttcaggatgc catcactgcc ttcatcaact
      481 ttgtgtccgc gatgctgaaa atgacggtga ctcgtcaaaa ttttgatgtc atcgtggctg
      541 agatcgagag gcttgcttca accagcgtat ctgtccgaac tgaagaggca aaggttgccg
      601 atgaagagct gatgctgttc gggctagatc atagagggcc acagcagctt gatatttcta
      661 atgctaaagg gataatgaaa gctgcagata ttcaggcaac tcatgacgtt catttggcac
      721 caggcgtcgg taatatcgat cctgagatct acaatgaggg acggttcatg tttatgcagc
      781 acaaaccact cgcggcagat cagtcgtatt tcacgttgga gactgcggat tatttcaaga
      841 tctacccaac atatgatgag catgatagca gaatggctga tcagaagcag tcagggttga
      901 tattgtgcac taaagatgag gtgttggctg agcaaactat ttttaagcta gatgcccctg
      961 atgataaaac cgttcatcta ttggatcgcg acgacgatca cgttgttgct agatttacta
     1021 aggtgtttat agaagatgtg gcccccgggc atcatgctgc gcaaaggtct ggccaacgct
     1081 ctgtgcttga tgacctatat gccaacaccc aagtggtttc aatcacttct gctgctctga
     1141 agtgggtagt taaacacggt gtgtccgacg ggattgtgaa tagaaagaat gtcaaagtgt
     1201 gtgttggttt tgaccctttg tacactctat ctacgcataa tggagtgtct ttgtgcgccc
     1261 tgctgatgga tgaaaaactt tccgtgctga acagcgcatg tcgtatgacg ctgcgttcac
     1321 taatgaaaac tggacgtgat gctgatgcac atagggcttt tcagcgagtg ctctctcagg
     1381 gatacgcatc attaatgtgc tattatcacc cttcacggaa gttagcatat ggtgaggtgc
     1441 tttttctaga gcgatccagt gacatggtag acgggattaa actacagttg gacgcatcta
     1501 gacagtgtca tgagtgtccc gtgttgcagc agaaagtagt tgagttggaa aaacagatta
     1561 tcatgcagaa gtccattcag tcagatccta ccccaatggc gctgcaacca ctgttatctc
     1621 agttgcgtga actgtctagt gaggtcaccc gactccagat ggagttaagt cggactcagt
     1681 ccctgaatgc tcagttggaa gcggatgcta agtcagctca agcatgtagt ctggatatgt
     1741 atttgagaca ccacacctgc attaatggtc atacaaaaga agatgaactg cttgatgctg
     1801 tacgtgtcgc tccagatgtg aggaaagaaa tcatggaaaa gaggggcgaa gtgagaaggg
     1861 gctggtgcga acgtatctct aaggaagcgg ctgccaaatg ccaaactgtt attgatgact
     1921 tgactcagat gaatggaaag caggcacgag agataacaga attacgcgag tcagccgaga
     1981 attatgagaa gcagattgcg gaattggtgg gcactattac tcaaaaccag atgacgtatc
     2041 agcaagagct acaagctttg gtagcgaaga atgtggaact ggatacgatg aaccaacgtc
     2101 aggctaaatc attgcgtatt actccctccc ttctatcagc cactcctatc gattcagtcg
     2161 acggcgctgc tgacctgatt gatttctccg ttccaactga tgagctgtaa atgagctgtg
     2221 acgcagtgtt gccctaatcc cttaagcctt cccgacccct attcatcggg tcggcatggc
     2281 atctccacct cctcgcggtc cgacctgggc atccgaagga ggacgtcgtc cactcggatg
     2341 gctaagggag agctcaaaaa aaaggatccg gctgctaaca aagcccgaaa ggaagctgag
     2401 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2461 ttgaggggtt ttttgctgaa aggaggaact atatccggat cgagatcctc tagagtcgac
     2521 ctgcaggcat gcaagcttgt attctatagt gtcacctaaa tcgtatgtgt atgatacata
     2581 aggttatgta ttaattgtag ccgcgttcta acgacaatat gtacaagcct aattgtgtag
     2641 catctggctt actgaagcag accctatcat ctctctcgta aactgccgtc agagtcggtt
     2701 tggttggacg aaccttctga gtttctggta acgccgttcc gcaccccgga aatggtcagc
     2761 gaaccaatca gcagggtcat cgctagccag atcctctacg ccggacgcat cgtggccggc
     2821 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     2881 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     2941 cccgtggccg ggggactgtt gggcgccatc tccttgcacc attccttgcg gcggcggtgc
     3001 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
     3061 gtcgatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     3121 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     3181 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     3241 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     3301 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3361 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3421 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3481 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3541 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3601 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3661 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3721 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3781 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3841 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3901 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3961 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4021 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4081 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4141 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4201 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4261 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4321 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4381 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4441 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4501 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4561 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4621 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4681 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4741 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4801 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4861 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4921 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4981 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5041 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5101 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5161 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5221 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     5281 cgcgcgttgg ccgattcatt aatgcagggg gat
//
