LOCUS       sequence-39536-e       10440 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 10440)
  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 10440)
  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-M1-S1-S2-S4T1L
FEATURES             Location/Qualifiers
     source          1..10440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1521..1568
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3248..3295
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4714..4761
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(4829..4847)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             6191..6202
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      7555..7602
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(7670..7688)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7979..7998)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(8209..8228)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8328..8350
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8388..8406)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8474..8578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8579..8647,8648..9439)
                     /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(8797..8816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9610..10198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10099..10118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10352..10369
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgatcccg ggtaatacga ctcactatag ctattcgctg gtcagttatg gctcgcgctg
       61 cgttcctatt caagactgtt ggatttggtg gtctgcaaaa tgtgccaatt aatgatgagt
      121 tatcgtcaca tctacttcga gccggtaatt cgccatggca gctgactcag ttcttagatt
      181 ggataagtct tggaagagga ttagctacat cagctcttgt tccaaccgct ggttcaagat
      241 attatcagat gagttgttta ttgagtggta ccctccagat tccatttcgt cctaatcatc
      301 gatggggaga tattaggttt ctgcgtctag tgtggtcagc tcctacgctt gacgggttgg
      361 ttgttgcccc accgcaggtc ttagctcagc cagcgttaca ggcgcaggca gatcgagtgt
      421 atgattgtga tgattaccca ttcttagcac gtgacccgag atttaagcat cgagtgtatc
      481 aacaattgag cgccgtaact ctgcttaact tgacggggtt tggtccaatt tcctatgttc
      541 gagtagacga ggatatgtgg agtggggatg tgaaccagct tctcatgaac tacttcgggc
      601 atacgtttgc ggaaattgcg tatacattat gtcaagcttc agccaataga ccttgggagc
      661 atgatggtac gtatgcgagg atgactcaaa ttatactgtc cttattttgg ttatcatatg
      721 tcggtgtaat tcatcagcag aacacgtacc ggacgttcta tttccaatgc aatcggcgcg
      781 gtgatgctgc tgaagtatgg attctttcct gctcattaaa ccactccgcc cagattagac
      841 cgggtaatcg cagtctattc gtcatgccaa caagtccaga ttggaatatg gacgtcaatt
      901 tgattttaag ttcgacgttg acagggtgct tgtgttcggg ctctcagtta ccgcttattg
      961 acaataactc agtgcctgcg gtttcgcgta acatccatgg ttggactggc agagctggca
     1021 accagctgca tggtttccaa gtgcgacgaa tggtgactga attctgtgac agattaagac
     1081 gcgatggagt tatgactcaa gctcagcaaa atcaaattga agcgttggca gatcaaactc
     1141 aacagtttaa gagggataaa cttgaggcgt gggctaggga agatgatcag tataatcagg
     1201 ctaatccgaa ttctacgatg ttccgtacga agccatttac gaatgcgcaa tggggacgag
     1261 gaaataccgg agcgactagt gccgcaattg cagcccttat ctaatcgtct tggagtgagg
     1321 ggatcccccc acacccctca cgactgacca cacattcatc gggtcggcat ggcatctcca
     1381 cctcctcgcg gtccgacctg ggcatccgaa ggaggacgtc gtccactcgg atggctaagg
     1441 gagagctcaa aaaaaaggat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     1501 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     1561 gttttttgct gaaaggagga actatatccg gatcgagatc ctctagataa tacgactcac
     1621 tatagctatt cgcgcctatg gatgcatctc tcattacaga gatacggaaa atagtactcc
     1681 aactatctgt atcaagcaat ggctcccagt caaaagaaat cgaggaaatc aagaaacaag
     1741 tccaggtcaa cgttgatgat atcagggctg ccaatattaa actcgacgga cttggaagac
     1801 agattgctga catcagcaat agcatctcaa ccattgagtc aagattgggt gagatggata
     1861 atcgacttgt gggtatctcg agtcaggtca cgcaattatc taactcagtt agccagaaca
     1921 ctcagagcat atcctcattg ggtgacagaa tcaatgctgt cgaaccacga gttgacagtc
     1981 tggatacggt cacgtctaat ctcactggac gaacatccac tttggaggca gatgttggaa
     2041 gcttacggac agaactagca gcgctaacaa cacgggtgac aactgaggtt acaaggttag
     2101 atggtctaat caatagtggc cagaattcga ttggtgagct atccacaaga ctatccaatg
     2161 tggagacgtc tatggtgacg acggctggac ggggactgca gaaaaacgga aacaccttga
     2221 acgtcattgt aggtaatgga atgtggttta atagttctaa tcaattgcag ctcgaccttt
     2281 cggggcaatc aaaaggggtg ggatttgtcg gcacaggaat ggtggttaag attgatacta
     2341 attattttgc ttacaatagt aatggagaga ttacattggt gagtcaaatc aatgaattgc
     2401 catcgcgcgt atcaacactg gaatcagcga aaatcgattc agttttacct ccattaaccg
     2461 tacgcgaagc gagcggcgta cgtaccctga gctttggtta tgatacgagc gattttacaa
     2521 tcatcaactc cgtactgtcg ttacggtcac gtttgactct tccgacatac aggtaccctc
     2581 tggagctcga cacagcaaat aatagagtgc aggtggcaga tcgttttggc atgcgcacgg
     2641 gtacttggac gggacaattg caatatcagc acccacaatt gagttggaga gcaaatgtca
     2701 ctttgaattt gatgaaggtg gatgattggt tggtgttgag cttttctcag atgacgacta
     2761 actcaataat ggcagatggg aaatttctga ttaattttgt gtctgggtta tcttctggat
     2821 ggcagacggg ggatactgaa ccatcgtcaa ctattgatcc attgtctacg acatttgccg
     2881 cggtccaatt tctaaataac ggtcaacgca ttgatgcgtt taggatcatg ggagtatcgg
     2941 aatggacgga tggagaatta gagattaaga attatggtgg cacatacacc ggtcatactc
     3001 aagtatattg ggctccgtgg acgatcatgt atccatgcaa tgtgaggtga atctagcgcg
     3061 aaccctcggc acaaggggtc aatcatcggg tcggcatggc atctccacct cctcgcggtc
     3121 cgacctgggc atccgaagga ggacgtcgtc cactcggatg gctaagggag agctcaaaaa
     3181 aaaggatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     3241 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     3301 aggaggaact atatccggat cgagatcctc tagagtcgac taatacgact cactatagct
     3361 atttttgcct cttcccaaac gttgtcgcaa tggaggtgtg cttgcccaac ggtcatcaga
     3421 tcgtggactt gattaacaac gcttttgaag gtcgtgtatc aatctacagc gcgcaagagg
     3481 gatgggacaa aacaatctca gcacagccag atatgatggt atgtggtggt gccatcgttt
     3541 gcatgcattg tctaggtgtt gttggatctc tgcaacgcaa gctgaagcat ttgcctcacc
     3601 atagatgtaa tcaacagatt cgtcatcagg attacgtcga tgtacagttc gcagatcgtg
     3661 ttactgctca ctggaagcgg ggtatgctgt cctttgttgc gcagatgcac gcgatgatga
     3721 atgacgtatc accagaggat ctagaccgtg tgcgtactga gggaggttca ctagtggagt
     3781 taaactggct tcaggttgat ccaaattcaa tgtttagatc aatacactca agttggacag
     3841 atcctctgca ggtagtggat gatcttgaca ctaagctgga tcaatactgg acggccctga
     3901 atctgatgat tgattcatcc gacttggtgc ccaacttcat gatgagagac ccatcacatg
     3961 cattcaatgg tgtgagactg gagggagacg cccgccaaac tcaattctct aggactttcg
     4021 attcgagatc gagtttagaa tggggtgtga tggtttacga ttactctgag ttagagcatg
     4081 atccatcgaa gggccgtgct tacaggaagg aattggtgac gccagcacga gacttcggtc
     4141 attttggatt atcccactac tctagggcga ctaccccaat ccttggaaag atgccagctg
     4201 tattctcggg aatgttgact gggaattgta aaatgtatcc attcatcaaa ggaacggcta
     4261 agttaaagac agtgcgcaag ctagtggatt cagtcaatca tgcgtggggc gtcgagaaga
     4321 tcagatatgc gcttggacca ggtggcatga cgggatggta caacaggact atgcagcagg
     4381 cccccattgt actaactccc gctgctctca caatgttctc agacaccacc aagttcgggg
     4441 atttggatta tccggtgatg attggcgatc cgatgattct tggctaaaca cccccatctt
     4501 cacagcgccg ggcctgacca acctggtgtg acgtgggaca ggctccattc atcgggtcgg
     4561 catggcatct ccacctcctc gcggtccgac ctgggcatcc gaaggaggac gtcgtccact
     4621 cggatggcta agggagagct caaaaaaaag gatccggctg ctaacaaagc ccgaaaggaa
     4681 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     4741 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatcgag atcctctaga
     4801 gtcgacctgc aggcatgcaa gcttgtattc tatagtgtca cctaaatcgt atgtgtatga
     4861 tacataaggt tatgtattaa ttgtagccgc gttctaacga caatatgtac aagcctaatt
     4921 gtgtagcatc tggcttactg aagcagaccc tatcatctct ctcgtaaact gccgtcagag
     4981 tcggtttggt tggacgaacc ttctgagttt ctggtaacgc cgttccgcac cccggaaatg
     5041 gtcagcgaac caatcagcag ggtcatcgct agctaatacg actcactata gctattcgcg
     5101 gtcatggctt acatcgcagt tcctgcggtg gtggattcac gttcaagtga ggctattgga
     5161 ctgctagaat cgtttggagt agacgctggg gctgatgcga atgacgtttc atatcaagat
     5221 catgactatg tgttggatca gttacagtat atgttagatg gatatgaggc tggcgacgtt
     5281 atcgatgcac tcgtccacaa gaattggtta catcactccg tctattgctt gttgccaccc
     5341 aaaagtcaac tactagagta ttggaaaagt aatccttcag tgataccgga caacgttgat
     5401 cgtcggcttc gtaaacgact aatgctaaag aaagatctca gaaaagatga tgaatacaat
     5461 caactagcgc gtgctttcaa gatatcggat gtctacgcac ctctcatctc atccacgacg
     5521 tcaccgatga caatgatcca gaacttgaat caaggcgaga tcgtgtacac cacgacggac
     5581 agggtaattg gggctagaat cttgttatat gctcctagaa agtactatgc gtcaactcta
     5641 tcatttacta tgactaagtg catcattccg tttggcaaag aggtgggtcg tgttcctcac
     5701 tctagattta atgttggcac atttccatca attgctaccc cgaaatgttt tgtcatgagt
     5761 ggggttgata ttgagtccat cccaaatgaa ttcatcaagt tgttttacca gcgcgtcaag
     5821 agtgttcacg ccaatatact aaatgacata tcacctcaga tcgtctctga catgataaac
     5881 agaaagcgtt tgcgcgttca tactccatca gatcgtcgag ccgcgcagtt gatgcatttg
     5941 ccctaccatg ttaaacgagg agcgtctcac gtcgacgttt acaaggtgga tgttgtagac
     6001 gtgttgtttg aggtagtgga tgtggccgat gggttgcgca acgtatctag gaaactaact
     6061 atgcataccg ttccggtatg tattcttgaa atgttgggta ttgagattgc ggactattgc
     6121 attcgtcaag aggatggaat gttcacagat tggttcctac ttttaaccat gctatctgat
     6181 ggcttaactg atagaaggac gcattgtcaa tacttgatta atccgtcaag tgtgcctcct
     6241 gatgtgatac ttaacatctc aattactgga tttataaata ggcatacaat cgatgtcatg
     6301 cctgatatat atgacttcgt taaacccatt ggcgctgtgc tgcctaaggg atcatttaaa
     6361 tcaacaatta tgagagttct tgattcaata tcaatattag gagtccagat catgccgcgc
     6421 gcgcatgtag ttgactcgga tgaggtgggc gagcaaatgg agcctacgtt tgagcatgcg
     6481 gttatggaga tatacaaagg gattgctggc gttgactcgc tggatgatct catcaagtgg
     6541 gtgctgaact cggatctcat tccgcatgat gacaggcttg gccaattatt tcaagcgttt
     6601 ctgcctctcg caaaggactt gttagctcca atggccagaa agttttatga taactcaatg
     6661 agtgagggta gattgctgac attcgctcat gccgacagtg agttgctgaa cgcaaattac
     6721 tttggtcatt tattgcgact aaaaatacca tatattacag aggttaatct gatgattcgc
     6781 aagaatcgtg agggtggaga gctatttcag cttgtgttat cgtatctata taaaatgtat
     6841 gctactagcg cgcagcctaa atggtttgga tcattattgc gattgttaat atgtccctgg
     6901 ttacatatgg agaaattaat aggagaagca gacccggcat ctacgtcggc tgaaattgga
     6961 tggcatatcc ctcgtgaaca gctgatgcaa gatggatggt gtggatgtga agatggattc
     7021 attccctatg ttagcatacg tgcgccaaga ctggttatgg aggagttgat ggagaagaac
     7081 tggggccaat atcatgccca agttattgtc actgatcagc ttgtcgtagg cgaaccgcgg
     7141 agggtatctg ccaaggctgt gatcaagggt aatcacttac cagttaagtt agtttcacga
     7201 tttgcatgtt tcacattgac ggcgaagtat gagatgaggc tctcgtgcgg ccatagcact
     7261 ggacgggggg ctgcatacaa tgcgagacta gctttccgat ctgacttggc gtgatccgtg
     7321 acatgcgtag tgtgacacct gcccctaggt caatgggggt agggggcggg ctaagactac
     7381 gtacgcgctt catcgggtcg gcatggcatc tccacctcct cgcggtccga cctgggcatc
     7441 cgaaggagga cgtcgtccac tcggatggct aagggagagc tcaaaaaaaa ggatccggct
     7501 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     7561 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     7621 tccggatcga gatcctctag agtcgacctg caggcatgca agcttgtatt ctatagtgtc
     7681 acctaaatcg tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg
     7741 acaatatgta caagcctaat tgtgtagcat ctggcttact gaagcagacc ctatcatctc
     7801 tctcgtaaac tgccgtcaga gtcggtttgg ttggacgaac cttctgagtt tctggtaacg
     7861 ccgttccgca ccccggaaat ggtcagcgaa ccaatcagca gggtcatcgc tagccagatc
     7921 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     7981 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     8041 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     8101 ttgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
     8161 taatgcagga gtcgcataag ggagagcgtc gatatggtgc actctcagta caatctgctc
     8221 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     8281 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     8341 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     8401 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     8461 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     8521 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     8581 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     8641 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     8701 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     8761 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     8821 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     8881 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     8941 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     9001 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     9061 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     9121 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     9181 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     9241 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     9301 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     9361 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     9421 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     9481 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     9541 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     9601 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     9661 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     9721 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     9781 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     9841 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     9901 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     9961 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    10021 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    10081 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    10141 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    10201 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
    10261 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
    10321 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
    10381 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagggggat
//
