LOCUS       sequence-372175-        8597 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 8597)
  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 8597)
  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-S1T3D
FEATURES             Location/Qualifiers
     source          1..8597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4087..4134
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5791..5838
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(5875..5893)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5876..5893)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6183..6202)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6412..6431)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6531..6553
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6591..6609)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6677..6781
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6782..6850,6851..7642)
                     /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(7000..7019)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7813..8401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8302..8321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8555..8572
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ggggatgggt aatacgactc
       61 actatagcta cacgttccac ttccacgaca atgtcatcca tgatactgac tcagtttgga
      121 ccgttcattg agagcatttc aggtatcact gatcaatcga atgacgtgtt tgaagatgca
      181 gcaaaagcat tctctatgtt tactcgcagc gatgtctaca aggcgctgga tgaaatacct
      241 ttctctgatg atgcgatgct tccaatccct ccaactatat atacgaaacc atctcacgat
      301 tcatattatt acattgatgc tctaaaccgt gtgcgtcgca aaacatatca gggccctgat
      361 gacgtgtacg tacctaattg ttctattgtt gaattgctgg agccacatga gactctgaca
      421 tcttatgggc ggttgtccga ggccatcgag aatcgtgcca aggatgggga cagccaagcc
      481 agaatcgcca caacgtatgg tagaatcgct gaatctcaag ctcgacagat taaggctcca
      541 ttggagaagt ttgtgttggc actattagtg gccgaagcag gggggtcttt atatgatcca
      601 gttttgcaga agtatgatga gattccagat ctatcgcata attgcccttt atggtgtttt
      661 agagagatct gtcgtcacat atctggtcca ttaccagatc gggcacctta tctttactta
      721 tctgcagggg ttttctggtt aatgtcacca cgaatgacgt ctgcaatccc tccgctacta
      781 tccgatcttg ttaatttagc tattttgcaa caaactgcgg gtttagatcc atcattagtg
      841 aaattgggag tacagatatg ccttcatgca gcagctagct caagttatgc atggtttatc
      901 ttaaagacta agtctatttt tcctcaaaac acgttgcaca gtatgtatga atctctagaa
      961 gggggatact gtcctaatct tgaatggtta gagcctagat cagactataa gttcatgtac
     1021 atgggagtca tgccattgtc cgctaagtat gctaggtcgg cgccgtccaa tgataagaaa
     1081 gcgcgggaac ttggcgagaa atatggactg agctcagtcg tcggtgagct tcgtaaacgg
     1141 acaaagacgt atgttaaaca tgactttgct tcagtgaggt acattcgtga cgctatggca
     1201 tgtactagcg gtattttctt ggtaagaaca cccaccgaaa cggtattgca agaatatacg
     1261 cagagtccgg agattaaggt tcccattccc cagaaagact ggacaggccc aataggtgaa
     1321 atcagaattc taaaagatac aacaagttcc atcgcgcgtt acttatatag aacatggtac
     1381 ttggcagcgg cgagaatggc ggctcaacca cgtacgtggg atccattgtt tcaagcgatt
     1441 atgagatctc aatacgtgac agctaggggt ggatctggcg cagcactccg cgaatctttg
     1501 tatgcgatca atgtgtcgtt acctgatttc aagggcttac cagtgaaggc agcaactaag
     1561 atattccagg cggcacaatt agcgaacttg ccgttctccc acacatcagt ggctatacta
     1621 gctgacactt caatgggatt gcgaaatcag gtgcagaggc ggccacgatc cattatgcca
     1681 ttaaatgtgc cccagcagca ggtttcggcg ccccatacat tgacagcgga ttacattaac
     1741 taccacatga atctatcaac cacgtctggt agtgcggtca ttgagaaggt gattccttta
     1801 ggtgtatacg cttcgagccc tcctaaccag tcgatcaaca ttgacatatc tgcgtgtgac
     1861 gctagtatta cttgggattt ctttctgtca gtgattatgg cggctataca cgaaggtgtc
     1921 gctagtagct ccattggaaa accatttatg ggggttcctg catccattgt aaatgatgag
     1981 tctgtcgttg gagtgagagc tgctaggccg atatcgggaa tgcagaacat gattcagcat
     2041 ctatcgaaac tatataaacg tggattttca tatagagtaa acgattcttt ttctccaggt
     2101 aacgatttta ctcatatgac taccactttc ccgtcaggtt caacagccac ctctactgag
     2161 catactgcta ataatagtac gatgatggaa actttcctga cagtatgggg acccgaacat
     2221 actgacgacc ctgacgtctt acgtttaatg aagtctttaa ctattcaaag aaattacgta
     2281 tgtcaaggtg atgatggatt aatgattatc gatgggacta ctgctggtaa ggtgaacagt
     2341 gaaactattc agaagatgct agaattaatc tcaaaatatg gtgaggaatt cggatggaaa
     2401 tatgacatag cgtacgatgg gactgccgaa tacttaaagc tatacttcat atttggctgt
     2461 cgaattccaa atcttagtcg ccatccaatc gtggggaaag aacgggcgaa ttcttcagca
     2521 gaggagccat ggccagcaat tctagatcag attatgggtg tcttctttaa tggtgttcat
     2581 gatgggttac agtggcagcg gtggatacgt tattcatggg ctctatgctg tgctttctca
     2641 cgtcaaagaa caatgattgg tgagagcgtg ggttaccttc aatatcctat gtggtctttt
     2701 gtctactggg gattaccact ggttaaagcg tttgggtcag acccatggat attttcttgg
     2761 tacatgccta ctggagatct gggaatgtat agttggatta gcttgatacg ccctctgatg
     2821 acaagatgga tggtggctaa tggttacgta actgacagat gctcacccgt attcgggaac
     2881 gcagattatc gcaggtgttt caatgaactt aaactatatc aaggttatta tatggcacaa
     2941 ttgcccagga atcctaagaa gtctggacga gcggcccctc gggaggtaag agaacaattc
     3001 actcaggcat tatccgacta tctaatgcaa aatccagaac tgaagtcacg tgtgctacgt
     3061 ggtcgtagtg agtgggagaa atatggagcg gggataattc acaatcctcc gtcattattc
     3121 gatgtgcccc ataaatggta tcagggtgcg caagaggcag caatcgctac gagagaagag
     3181 ctggcagaaa tggatgagac attaatgcgc gctcgaaggc acagctattc gaacttttca
     3241 aagttattag aggcgtatct gctcgtgaaa tggcgaatgt gcgaggcccg cgaaccgtcg
     3301 gttgatttgc gattaccatt atgtgcgggt attgacccat taaactcaga tccttttctc
     3361 aagatggtaa gcgttggacc aatgctccag agtacgagaa agtactttgc tcagacacta
     3421 ttcatggcaa agacggtgtc gggtcttgac gttaacgcga ttgatagcgc gttattacga
     3481 ctgcgaacat taggtgctga taagaaagca ttaacggcgc agttattaat ggtggggctt
     3541 caggagtcag aagcggacgc attggccggg aagataatgc tacaggatgt gaatactgtg
     3601 caattagcca gagtggttaa cttagctgtg ccagatactt ggatgtcgtt agactttgac
     3661 tctatgttca aacaccacgt caagctgctt cccaaagatg gacgtcatct aaatactgat
     3721 attcctcctc gaatgggatg gttacgggcc attttacgat tcttaggtgc cggaatggta
     3781 atgactgcga ctggagttgc tgtcgacatc tatctggagg atatacatgg cggtggtcgg
     3841 tcacttggac agagattcat gacttggatg cgacaggaag gacggtcagc gtgagtctac
     3901 catgggtcgt ggtgcgtcaa ctcatcgggt cggcatggca tctccacctc ctcgcggtcc
     3961 gacctgggca tccgaaggag gacgtcgtcc actcggatgg ctaagggaga gctcaaaaaa
     4021 aaggatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     4081 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     4141 ggaggaacta tatccggatc gagatcctct agagtcgacc tgcaggcatg caagctttaa
     4201 tacgactcac tatagctatt ggtcggatgg atcctcgcct acgtgaagaa gtagtacggc
     4261 tgataatcgc attaacgagt gataatggag tatcactgtc aaaagggctt gaatcaaggg
     4321 tctcggcgct cgagaagacg tctcaaatac actctgatac tatcctccgg atcacccagg
     4381 gactcgatga tgcaaacaaa cgaatcatcg ctcttgagca aagtcgggat gacttggttg
     4441 catcagtcag tgatgctcaa cttgcaatct ccagattgga aagctctatc ggagccctcc
     4501 aaacagttgt caatggactt gattcgagtg ttacccagtt gggtgctcga gtgggacaac
     4561 ttgagacagg acttgcagag ctacgcgttg atcacgacaa tctcgttgcg agagtggata
     4621 ctgcagaacg taacattgga tcattgacca ccgagctatc aactctgacg ttacgagtaa
     4681 catccataca agcggatttc gaatctagga tatccacatt agagcgcacg gcggtcacta
     4741 gcgcgggagc tcccctctca atccgtaata accgtatgac catgggatta aatgatggac
     4801 tcacgttgtc agggaataat ctcgccatcc gattgccagg aaatacgggt ctgaatattc
     4861 aaaatggtgg acttcagttt cgatttaata ctgatcaatt ccagatagtt aataataact
     4921 tgactctcaa gacgactgtg tttgattcta tcaactcaag gataggcgca actgagcaaa
     4981 gttacgtggc gtcggcagtg actcccttga gattaaacag tagcacgaag gtgctggata
     5041 tgctaataga cagttcaaca cttgaaatta attctagtgg acagctaact gttagatcga
     5101 catccccgaa tttgaggtat ccgatagctg atgttagcgg cggtatcgga atgagtccaa
     5161 attataggtt taggcagagc atgtggatag gaattgtctc ctattctggt agtgggctga
     5221 attggagggt acaggtgaac tccgacattt ttattgtaga tgattacata catatatgtc
     5281 ttccagcttt tgacggtttc tctatagctg acggtggaga tctatcgttg aactttgtta
     5341 ccggattgtt accaccgtta cttacaggag acactgagcc cgcttttcat aatgacgtgg
     5401 tcacatatgg agcacagact gtagctatag ggttgtcgtc gggtggtacg cctcagtata
     5461 tgagtaagaa tctgtgggtg gagcagtggc aggatggagt acttcggtta cgtgttgagg
     5521 ggggtggctc aattacgcac tcaaacagta agtggcctgc catgaccgtt tcgtacccgc
     5581 gtagtttcac gtgaggatca gaccaccccg cggcactggg gcatttcatc gggtcggcat
     5641 ggcatctcca cctcctcgcg gtccgacctg ggcatccgaa ggaggacgtc gtccactcgg
     5701 atggctaagg gagagctcaa aaaaaaggat ccggctgcta acaaagcccg aaaggaagct
     5761 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     5821 gtcttgaggg gttttttgct gaaaggagga actatatccg gatcaagctt gtattctata
     5881 gtgtcaccta aatcgtatgt gtatgataca taaggttatg tattaattgt agccgcgttc
     5941 taacgacaat atgtacaagc ctaattgtgt agcatctggc ttactgaagc agaccctatc
     6001 atctctctcg taaactgccg tcagagtcgg tttggttgga cgaaccttct gagtttctgg
     6061 taacgccgtc ccgcacccgg aaatggtcag cgaaccaatc agcagggtca tcgctagcca
     6121 gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     6181 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     6241 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     6301 ctccttgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
     6361 ttcttaatgc aggagtcgca taagggagag cgtcgaatgg tgcactctca gtacaatctg
     6421 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     6481 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     6541 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     6601 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     6661 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6721 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6781 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6841 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6901 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6961 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7021 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7081 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7141 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7201 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7261 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7321 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7381 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7441 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7501 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7561 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7621 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     7681 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     7741 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7801 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7861 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7921 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     7981 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     8041 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     8101 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     8161 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     8221 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     8281 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     8341 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     8401 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     8461 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     8521 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     8581 agagcgccca atacgca
//
