LOCUS       sequence-39531-e        9413 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 9413)
  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 9413)
  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-L1-M2T1L
FEATURES             Location/Qualifiers
     source          1..9413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4044..4091
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      6528..6575
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(6643..6661)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6644..6661)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6952..6971)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7182..7201)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7301..7323
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7361..7379)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7447..7551
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7552..7620,7621..8412)
                     /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(7770..7789)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8583..9171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9072..9091
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9325..9342
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgatcccg ggtaatacga ctcactatag ctacacgttc cacgacaatg tcatccatga
       61 tactgactca gtttagaccg ttcattgaaa gcatttcagg tatcactgac caatcgaatg
      121 acgtgtttga agatgcagca aaagcattct ctatgtttac tcgcagcgac gtctacaagg
      181 cgctggatga aatacctttc tctgatgatg cgatgcttcc catccctcca actatatata
      241 ctaaaccatc tcacgattca tattattaca ttgatgctct aaaccgtgtg cgtcgcaaaa
      301 catatcaggg ccctgatgac gtgtacgtac ctaattgttc tattgttgaa ttgctggagc
      361 cacatgagac tctgacatct tatgggcgtt tgtctgaggc catcgagaat cgtgccaagg
      421 atggagacag ccaagccaga atcgcaacga cgtatggtag aatcgctgaa tctcaagctc
      481 gacagattaa ggctccattg gagaagtttg tgttggcact actagtgtcc gaagcggggg
      541 ggtctttata tgatccagtt ctgcagaagt atgatgagat tccagatcta tcgcataatt
      601 gccctttatg gtgttttaga gaaatctgtc gtcatatatc tggtccatta ccagaccgag
      661 caccttatct ttacttatct gcaggggttt tctggttaat gtcaccacga atgacgtctg
      721 caatccctcc gttactatcc gatcttgtta atttagctat tttacaacag actgcgggtt
      781 tggatccatc attagtgaaa ttgggagtgc agatatgcct tcatgcggca gctagctcaa
      841 gttatgcatg gtttatctta aagactaagt ctatttttcc tcaaaacacg ttacatagta
      901 tgtatgaatc tctagaaggg gggtattgtc ctaaccttga atggttagag cctagatcgg
      961 actataaatt catgtacatg ggagtcatgc cattgtccac taagtatgct aggtcggcgc
     1021 cgtcaaatga taagaaagcg cgggaacttg gcgagaaata tggactgagc tcagtcgtca
     1081 gtgagcttcg taaacggaca aagacttatg ttaaacatga ctttgcttcg gtaaggtaca
     1141 ttcgtgacgc tatggcatgt actagcggta ttttcttggt aagaacaccc accgaaacgg
     1201 tattgcaaga atatactcag agtccggaga ttaaggttcc catcccccaa aaagattgga
     1261 caggcccagt aggtgagatc agaattctaa aagacacaac aagctccatc gcgcgctact
     1321 tatatagaac atggtactta gcagcggcaa gaatggcggc tcaaccacgt acgtgggatc
     1381 cattgtttca ggcgattatg agatctcaat acgtgacagc taggggtgga tctggcgcag
     1441 cactccgcga gtctctgtac gcaattaatg tgtcgttacc tgattttaag ggcttaccag
     1501 tgaaggcagc aactaagata tttcaggcgg cacaattagc gaacttgccg ttctcccaca
     1561 catcagtggc tatactagct gacacttcga tgggattgcg aaaccaggtg cagaggcggc
     1621 cacgatccat tatgccctta aatgtgcccc aacagcaggt ttcggcgcct catacattga
     1681 ccgcggatta catcaattac cacatgaacc tatcaaccac gtctggtagc gcggtcattg
     1741 agaaagtgat tcccttaggt gtatacgctt cgagccctcc taaccaatcg attaacattg
     1801 acatatctgc atgtgacgca agtattactt gggatttctt tctatccgtg attatggcgg
     1861 ctatacacga aggtgtcgct agtagctcca ttggaaaacc gtttatgggg gttcctgcat
     1921 ccattgtaaa tgatgagtct gtcgttggag tgagagctgc taggccgata tcgggaatgc
     1981 agaacatgat tcagcatcta tcaaaactgt ataaacgtgg attttcatac agagtgaacg
     2041 actctttttc tccaggtaac gactttaccc atatgactac cactttcccg tcaggttcaa
     2101 cagccacctc tactgagcat actgctaata atagtacgat gatggaaact ttcctgacag
     2161 tatggggacc cgaacatact gacgaccccg acgtcttacg tctaatgaag tctttaacta
     2221 ttcaaaggaa ttacgtgtgt caaggtgatg atggattgat gattatcgat gggaatactg
     2281 ctggtaaggt gaacagtgaa actattcaga agatgctgga gttaatctca aaatatggtg
     2341 aggaatttgg atggaaatat gacatagcgt acgatgggac tgccgaatac ctaaagctgt
     2401 atttcatatt tggctgtcga attccaaatc ttagtcgtca tccaatcgtg gggaaagaac
     2461 gggcgaattc ttcagcagag gagccatggc cagcaattct agaccagatt atgggtatct
     2521 tctttaatgg cgttcatgac gggttacagt ggcagcggtg gatacgttat tcatgggctc
     2581 tatgctgtgc tttctcacgt caaagaacaa tgattggtga gagcgtaggt taccttcaat
     2641 atcctatgtg gtcttttgtc tactggggat taccactggt taaagtgttc gggtcagacc
     2701 catggatatt ttcttggtac atgcctactg gggacctggg aatgtatagt tggattagcc
     2761 tgatacgccc tctaatgaca agatggatgg tggctaatgg ttacgcaact gacagatgct
     2821 cacccgtatt cgggaacgca gattatcgca ggtgtttcaa tgaaattaaa ctatatcaag
     2881 ggtattatat ggcacaattg cctaggaatc ctacgaaatc tggacgagcg gcccctcggg
     2941 aggtaagaga acaattcact caggcattat ccgattatct gatgcaaaat ccagagctga
     3001 agtcacgtgt gctacgtggt cgtagtgagt gggagaagta tggagcgggg ataattcaca
     3061 atcctccatc attattcgat gtgccccata agtggtatct gggtgcgcaa gaggcggcaa
     3121 ccgctacgag agaagagctg gcagaaatgg atgagacatt aatgcgcgct cgaaggcaca
     3181 gttattcaag cttttcaaag ttgttggagg catacctgct cgtgaaatgg cgaatgtgcg
     3241 aggcccgcga accgtcggtt gatttgcgat taccattatg tgcgggtatt gacccactaa
     3301 actcagatcc ttttctcaag atggtaagcg ttgggccaat gctccagagt acgagaaagt
     3361 actttgctca gacactattc atggcaaaga cggtgtcggg tctcgacgtt aacgcgattg
     3421 acagcgcgtt attacgactg cgaacattgg gtgctgataa gaaagcatta acagcgcagt
     3481 tattaatggt ggggcttcag gagtcagaag ctgacgcgtt ggctgggaag ataatgctac
     3541 aagatgtaag tactgtgcaa ttagccagag tggtcaactt agctgtgcca gatacttgga
     3601 tgtcgttgga ctttgactct atgttcaaac accatgttaa gttgcttccc aaagatggac
     3661 gtcatctaaa tactgatatt cctcctcgaa tgggatggtt acgggccatt ttacgattct
     3721 taggtgctgg aatggtaatg actgcgactg gagttgctgt cgacatatat ctggaggata
     3781 tacacggcgg tggtcgagca cttggacaga gattcatgac ttggatgcga caggaaggac
     3841 ggtcagcgtg agtctaccat gggtcgtggt gcgtcaactc atcgggtcgg catggcatct
     3901 ccacctcctc gcggtccgac ctgggcatcc gaaggaggac gtcgtccact cggatggcta
     3961 agggagagct caaaaaaaag gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     4021 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     4081 ggggtttttt gctgaaagga ggaactatat ccggatcgag atcctctaga gtcgacctgc
     4141 aggcatgcta atacgactca ctatagctaa tctgctgacc gtcactctgc aaagatgggg
     4201 aacgcttcct ctatcgttca gacgatcaac gttactggag atggcaatgt attcaaaccc
     4261 tcagctgaga cttcatccac cgctgtaccg tcactgagtc tatcacctgg aatgttaaat
     4321 cctggaggag taccatggat tgcgattgga gatgaaacat ccgttacttc accgggtgca
     4381 ttgcggcgaa tgacttcgaa ggatatccca gagacagcaa taatcaatac ggataattca
     4441 tcaggcgcgg tgccaagtga atcagcatta gtgccctata atgatgagcc attggtagtg
     4501 gtgacggagc acgctatcgc aaacttcacc aaagccgaga tggcacttga gttcaatcgt
     4561 gagttcctcg ataaattgcg cgtgctgtca gtgtcaccaa aatattccga ccttctgacg
     4621 tatgttgatt gttacgttgg tgtgtcggcc cgtcaggccc taaataattt ccagaagcag
     4681 gtgcctgtga ttacacctac tagacaaaca atgtatgttg actccataca ggcggccttg
     4741 aaagccttag agaaatggga aattgatttg agagtggctc agacactgtt gcctacaaat
     4801 gtcccaattg gggaggtctc ttgtccaatg cagtcagtag tgaaactatt agacgatcag
     4861 ctgccggacg atagccttat acggaggtat cctaaggagg ccgctgttgc tttggctaaa
     4921 aggaatggag ggatacagtg gatggacgta tcagaaggaa ctgtgatgaa tgaggccgtg
     4981 aatgctgtcg cagcaagtgc cctggcacct tccgcatcag ccccaccctt agaagagaaa
     5041 tcgaaattga ctgagcaagc gatggatctt gtaactgcag ctgaacctga gataatcgca
     5101 tctctcgtac cagttccagc gcccgtgttt gccattccac ctaagccagc cgactataat
     5161 gtgcgtactt tgaagatcga tgaggccaca tggttacgaa tgatcccaaa aactatgggt
     5221 acgccttttc aaatccaagt gactgacaac acaggaacta actggcacct taacttgaga
     5281 ggagggacac gcgtagtgaa cttggatcag attgctccga tgagattcgt tctggatctg
     5341 gggggaaaaa gctataagga gacaagttgg gatccaaatg gtaagaaggt tgggtttatc
     5401 gtatttcaat ctaagattcc ttttgagctt tggaccgccg cctcacagat tggccaagct
     5461 acagtggtta actatgttca gctatatgct gaagacagct catttaccgc ccagtccatc
     5521 atcgccacta catcgttggc ttataattat gaacctgagc aattgaataa gactgaccct
     5581 gagatgaact attacctttt agcgactttc atagactcag ctgctataac accgacaaac
     5641 atgacacagc ctgatgtttg ggatgctctg ttgacgatgt ctccactgtc cgctggagag
     5701 gtgactgtga agggtgcggt agtaagcgag gtggtcccag cggagttgat agggagctat
     5761 actccagagt cactaaatgc ctcacttccg aatgacgctg ctagatgtat gatcgataga
     5821 gcttcgaaga tagccgaggc tataaaaatt gatgatgacg ccgggccaga cgaatactct
     5881 cccaactctg taccgatcca aggtcagctt gctatttctc aacttgagac tgggtatggt
     5941 gtacggatat tcaatcccaa aggaattctc tcaaaaatcg catctagagc tatgcaggct
     6001 tttattggtg atccaagtac aattattacg caggcagcac cggtgctgtc agataagaat
     6061 aattggattg cattggcaca aggagttaag actagtttgc gtactaaaag tttatcggcg
     6121 ggggtgaaga cggcggtgag taagctgagc tcatccgagt ctattcaaaa ttggactcaa
     6181 ggattcttgg ataaggtatc gacgcacttt ccagcgccta agcccgactg tccgaccaac
     6241 ggagatggca gtgaaccgtc tgctcggcga gtgaagcgcg actcatacgc aggagtggtt
     6301 aagcgtgggt atacacgcta agctgctcgc cctggtgacg cggggttaag ggtgcaggca
     6361 catcatcggg tcggcatggc atctccacct cctcgcggtc cgacctgggc atccgaagga
     6421 ggacgtcgtc cactcggatg gctaagggag agctcaaaaa aaaggatccg gctgctaaca
     6481 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     6541 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     6601 cgagatcctc tagagtcgac ctgcaggcat gcaagcttgt attctatagt gtcacctaaa
     6661 tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta acgacaatat
     6721 gtacaagcct aattgtgtag catctggctt actgaagcag accctatcat ctctctcgta
     6781 aactgccgtc agagtcggtt tggttggacg aaccttctga gtttctggta acgccgttcc
     6841 gcaccccgga aatggtcagc gaaccaatca gcagggtcat cgctagccag atcctctacg
     6901 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     6961 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
     7021 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcacc
     7081 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
     7141 ggagtcgcat aagggagagc gtcgatatgg tgcactctca gtacaatctg ctctgatgcc
     7201 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     7261 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     7321 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     7381 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     7441 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7501 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7561 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7621 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7681 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     7741 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     7801 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     7861 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     7921 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     7981 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     8041 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     8101 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     8161 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     8221 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     8281 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     8341 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     8401 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     8461 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     8521 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8581 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     8641 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     8701 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     8761 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     8821 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     8881 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     8941 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     9001 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     9061 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     9121 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     9181 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     9241 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     9301 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     9361 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagggg gat
//
