LOCUS       sequence-39539-e        9688 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 9688)
  AUTHORS   Kobayashi T, Ooms LS, Ikizler M, Chappell JD, Dermody TS
  TITLE     An improved reverse genetics system for mammalian orthoreoviruses.
  JOURNAL   Virology. 2010 Mar 15;398(2):194-200. Epub 2009 Dec 29.
  PUBMED    20042210
REFERENCE   2  (bases 1 to 9688)
  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: "Virology.";
            date: "2010-03-15"; volume: "398(2)"; pages: "194-200. Epub 2009 Dec
            29"
COMMENT     Sequence Label: pT7-M2-S2-S3-S4T3D
FEATURES             Location/Qualifiers
     source          1..9688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1702..1749
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3170..3217
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4623..4670
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      7083..7130
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(7227..7246)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7457..7476)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7576..7598
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7636..7654)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7722..7826
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7827..7895,7896..8687)
                     /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(8045..8064)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8858..9446
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9347..9366
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9600..9617
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctaggttgtt ctctatcttt gtgatgtggc tctgaaacaa accaggtgtg attacagtaa
       61 caatttcaat ttaaattccg atataaattt caatgagagg aaaattatta atcttcctca
      121 tagttattcg cacacaaaag atcctaaaaa ttcttctttc tttttgtgtg tccggggtcg
      181 gcatggctct agataatacg actcactata gctattcgct ggtcagttat ggctcgcgct
      241 gcgttcctat tcaagactgt tgggtttggt ggtctgcaaa atgtgccaat taacgacgaa
      301 ctatcttcac atctactccg agctggtaat tcaccatggc agttaacaca gtttttagac
      361 tggataagcc ttgggagggg tttagctaca tcggctctcg ttccgacggc tgggtcaaga
      421 tactatcaaa tgagttgcct tctaagtggc actctccaga ttccgttccg tcctaaccac
      481 cgatggggag acattaggtt cttacgctta gtgtggtcag ctcctactct cgatggatta
      541 gtcgtagctc caccacaagt tttggctcag cccgctttgc aagcacaggc agatcgagtg
      601 tacgactgcg atgattatcc atttctagcg cgtgatccaa gattcaaaca tcgggtgtat
      661 cagcaattga gtgctgtaac tctacttaac ttgacaggtt ttggcccgat ttcctacgtt
      721 cgagtggatg aagatatgtg gagtggagat gtgaaccagc ttctcatgaa ctatttcggg
      781 cacacgtttg cagagattgc atacacattg tgtcaagcct cggctaatag gccttgggaa
      841 tatgacggta catatgctag gatgactcag attgtgttat ccttgttctg gctatcgtat
      901 gtcggtgtaa tccatcagca gaatacgtat cggacattct attttcagtg taatcggcga
      961 ggtgacgccg ctgaggtgtg gattctttct tgttcgttga accattccgc acaaattaga
     1021 ccgggtaatc gtagcttatt cgttatgcca actagcccag attggaacat ggacgtcaat
     1081 ttgatcctga gttcaacgtt gacggggtgt ttgtgttcgg gttcacagct gccactgatt
     1141 gacaataatt cagtacctgc agtgtcgcgt aacatccatg gctggactgg tagagctggt
     1201 aaccaattgc atgggttcca ggtgagacga atggtgactg aattttgtga caggttgaga
     1261 cgcgatggtg tcatgaccca agctcagcag aatcaagttg aagcgttggc agatcagact
     1321 caacagttta agagggacaa gctcgaaacg tgggcgagag aagacgatca atataatcag
     1381 gctcatccca actccacaat gttccgtacg aaaccattta cgaatgcgca atggggacga
     1441 ggtaatacgg gggcgactag tgccgcgatt gcagccctta tctgatcgtc ttggagtgag
     1501 ggggtccccc cacacccctc acgactgacc acacattcat cgggtcggca tggcatctcc
     1561 acctcctcgc ggtccgacct gggcatccga aggaggacgt cgtccactcg gatggctaag
     1621 ggagagctca aaaaaaagga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     1681 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1741 ggttttttgc tgaaaggagg aactatatcc ggatcgagat cctctagagt cgactaatac
     1801 gactcactat agctaaagtc acgcctgtcg tcgtcactat ggcttcctca ctcagagctg
     1861 cgatctccaa gatcaagagg gatgacgtcg gtcagcaagt ttgtcctaat tatgtcatgc
     1921 tgcggtcctc tgtcacaaca aaggtggtac gaaatgtggt tgagtatcaa attcgtacgg
     1981 gcggattctt ttcgtgctta gctatgctaa ggccactcca gtacgctaag cgtgagcgtt
     2041 tgcttggtca gaggaatctg gaacgtatat cgactaggga tatccttcag actcgtgatt
     2101 tacactcact atgtatgcca actcctgatg cgccaatgtc taatcatcaa gcatccacca
     2161 tgagagagct gatttgcagt tacttcaagg tcgatcatgc ggatgggttg aaatatatac
     2221 ccatggatga gagatactct ccgtcatcac ttgccagatt gtttaccatg ggcatggctg
     2281 ggctgcacat taccactgag ccatcttata agcgtgttcc gattatgcac ttagctgcgg
     2341 acttggactg tatgacgctg gctctacctt acatgattac gcttgatggt gatactgtgg
     2401 ttcctgtcgc tccaacactg tcagcggaac agcttctgga cgacggactc aaaggattag
     2461 catgcatgga tatctcctat ggatgtgagg tggacgcgaa tagccggccg gctggtgatc
     2521 agagtatgga ctcttcacgc tgcatcaacg agttgtattg cgaggagaca gcagaagcca
     2581 tctgtgtgct taagacatgc cttgtgttaa attgcatgca gtttaaactt gagatggatg
     2641 acctagcaca taacgctgct gagctggaca agatacagat gatgataccc ttcagtgagc
     2701 gtgtttttag gatggcctcg tcctttgcga ctattgatgc ccagtgtttt aggttttgcg
     2761 tgatgatgaa ggataaaaat ctgaaaatag atatgcgtga aacgacgaga ctgtggactc
     2821 gttcagcatc agatgattct gtggccacgt catctttaag tatttccctg gaccggggtc
     2881 gatgggtggc ggctgacgcc agtgatgcta gactgctggt ttttccgatt cgcgtgtaat
     2941 gggtgagtga gctgatgtgg tcgccaagac atgtgccggt gtcttggtgg tgggtgacgc
     3001 ctaatcatcg ggtcggcatg gcatctccac ctcctcgcgg tccgacctgg gcatccgaag
     3061 gaggacgtcg tccactcgga tggctaaggg agagctcaaa aaaaaggatc cggctgctaa
     3121 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     3181 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     3241 atcaagcttt aatacgactc actatagcta tttttgcctc ttcccagacg ttgtcgcaat
     3301 ggaggtgtgc ttgcccaacg gtcatcaggt cgtggacttg attaacaacg cttttgaagg
     3361 tcgtgtatca atctacagcg cgcaagaggg atgggacaaa acaatctcag cacagccaga
     3421 tatgatggta tgtggtggcg ccgtcgtttg catgcattgt ctaggtgttg tcggatctct
     3481 acaacgcaag ctgaagcatt tgcctcacca tagatgtaat caacagatcc gtcatcagga
     3541 ttacgtcgat gtacagttcg cagaccgtgt tactgctcac tggaagcggg gtatgctgtc
     3601 cttcgttgcg cagatgcacg agatgatgaa tgacgtgtcg ccagatgacc tggatcgtgt
     3661 gcgtactgag ggaggttcac tagtggagct gaactggctt caggttgacc caaattcaat
     3721 gtttagatca atacactcaa gttggacaga tcctttgcag gtggtggacg accttgacac
     3781 taagctggat cagtactgga cagccttaaa cctgatgatc gactcatccg acttgatacc
     3841 caactttatg atgagagacc catcacacgc gttcaatggt gtgaaactgg ggggagatgc
     3901 tcgtcaaacc caattctcca ggacttttga ttcgagatcg agtttggaat ggggtgtgat
     3961 ggtttatgat tactctgagc tggagcatga tccatcgaag ggccgtgctt acagaaagga
     4021 attggtgacg ccagctcgag atttcggtca ctttggatta tcccattatt ctagggcgac
     4081 taccccaatc cttggaaaga tgccggccgt attctcagga atgttgactg ggaactgtaa
     4141 aatgtatcca ttcattaaag gaacggctaa gctgaagaca gtgcgcaagc tagtggaggc
     4201 agtcaatcat gcttggggtg tcgagaagat tagatatgct cttgggccag gtggcatgac
     4261 gggatggtac aataggacta tgcaacaggc ccccattgtg ctaactcctg ctgctctcac
     4321 aatgttccca gataccatca agtttgggga tttgaattat ccagtgatga ttggcgatcc
     4381 gatgattctt ggctaaacac ccccatcttc acagcgccgg gcttgaccaa cctggtgtga
     4441 cgtgggacag gcttcattca tcgggtcggc atggcatctc cacctcctcg cggtccgacc
     4501 tgggcatccg aaggaggacg tcgtccactc ggatggctaa gggagagctc aaaaaaaagg
     4561 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     4621 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     4681 gaactatatc cggatcgcta gctaatacga ctcactatag ctaatctgct gaccgttact
     4741 ctgcaaagat ggggaacgct tcctctatcg ttcagacgat caacgtcact ggagatggca
     4801 atgtatttaa accatcagct gaaacttcat ctaccgctgt accatcgtta agcttatcac
     4861 ctggaatgct gaatcccgga ggggtaccat ggattgctgt tggagatgag acatctgtga
     4921 cttcaccagg cgcattacgt cgaatgacgt caaaggacat cccggaaacg gcaataatca
     4981 acacagacaa ttcatcaggc gccgtgccaa gcgaatcagc cttggtgccc tacatcgatg
     5041 agccgctggt agtggttaca gagcatgcta ttaccaactt caccaaagct gagatggcac
     5101 ttgaattcaa tcgtgagttc cttgacaaga tgcgtgtgct gtcagtgtca ccaaaatatt
     5161 cggatcttct gacctatgtt gactgctacg tcggtgtgtc tgctcgtcag gctttaaaca
     5221 attttcagaa acaagtgcct gtgattacac ctactaggca gacgatgtat gtcgactcga
     5281 tacaagcggc cttgaaagct ttagaaaagt gggagattga tctgagagtg gctcaaacgt
     5341 tgctgcctac gaacgttccg attggagaag tctcttgtcc aatgcagtcg gtagtgaaac
     5401 tgctggatga tcagctgcca gatgacagcc tgatacggag gtatcccaag gaagccgccg
     5461 tcgctttggc taaacgaaac gggggaatac aatggatgga cgtatcagaa ggcaccgtga
     5521 tgaacgaggc tgtcaacgct gttgcagcta gtgcactggc accttcagca tcagccccac
     5581 ccttagaaga gaagtcaaag ttaaccgaac aagcgatgga tctcgtgacc gcggctgagc
     5641 ctgagataat tgcctcactc gcgccagttc ccgcacccgt gtttgccata ccacctaaac
     5701 cagcagatta taatgtgcgt actctgagga tcgacgaggc cacttggctg cgaatgattc
     5761 caaaatcaat gaacacacct tttcaaatcc aggtgactga taacacagga actaattggc
     5821 atctcaattt gagggggggg actcgtgtag tgaatctgga ccaaatcgct ccgatgcggt
     5881 ttgtattaga tttaggggga aagagttata aagagacgag ctgggatcca aacggcaaga
     5941 aggtcggatt catcgttttt caatcgaaga taccattcga actttggact gctgcttcac
     6001 agatcggtca agccacggtg gttaactatg tccaactata cgctgaagac agctcattta
     6061 ccgcgcagtc tatcattgct actacctctt tggcttataa ctatgagcct gagcagttga
     6121 ataagactga ccctgagatg aattattatc ttttggcgac ctttatagac tcagccgcta
     6181 taacgccaac gaatatgaca cagcctgatg tttgggatgc cttgctgacg atgtccccac
     6241 tatcagctgg cgaggtgaca gtgaagggtg cggtagtgag tgaagtagtc cctgcagact
     6301 tgataggtag ctacactcca gaatccctaa acgcctcact tccgaatgat gctgctagat
     6361 gcatgatcga tagagcttcg aagatagccg aagcaatcaa gattgatgat gatgctggac
     6421 cagatgaata ttccccaaac tctgtaccaa ttcaaggtca gcttgctatc tcgcaactcg
     6481 aaactggata tggtgtgcga atattcaacc ctaaagggat cctttctaaa attgcatcta
     6541 gggcaatgca ggctttcatt ggtgacccga gcacaatcat cacgcaggcg gcgccagtgt
     6601 tatcagacaa gaataattgg attgcattgg cacagggagt gaaaactagt ctgcgtacta
     6661 aaagtctatc agcgggagtg aagactgcag tgagtaagct gagctcatct gagtctatcc
     6721 agaattggac tcaaggattc ttggataaag tgtcagcgca ttttccagca ccaaagcccg
     6781 attgtccgac tagcggagat agtggtgaat cgtctaatcg ccgagtgaag cgcgactcat
     6841 acgcaggagt ggtcaaacgt gggtacacac gttaggccgc tcgccctggt gacgcggggt
     6901 taagggatgc aggcaaatca tcgggtcggc atggcatctc cacctcctcg cggtccgacc
     6961 tgggcatccg aaggaggacg tcgtccactc ggatggctaa gggagagctc aaaaaaaagg
     7021 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     7081 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     7141 gaactatatc cggatcgcta gccagatcct ctacgccgga cgcatcgtgg ccggcatcac
     7201 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     7261 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     7321 ggccggggga ctgttgggcg ccatctcctt gcaccattcc ttgcggcggc ggtgctcaac
     7381 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga
     7441 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     7501 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     7561 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     7621 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     7681 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     7741 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     7801 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     7861 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     7921 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     7981 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     8041 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     8101 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     8161 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     8221 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     8281 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     8341 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     8401 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     8461 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     8521 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     8581 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     8641 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     8701 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     8761 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     8821 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     8881 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     8941 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     9001 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     9061 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     9121 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     9181 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     9241 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     9301 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     9361 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     9421 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     9481 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     9541 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     9601 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     9661 gttggccgat tcattaatgc agggggat
//
