LOCUS       sequence-39538-e        9542 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9542)
  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 9542)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-L3-M1T3D
FEATURES             Location/Qualifiers
     source          1..9542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4096..4143
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             5324..5335
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      6688..6735
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(6772..6790)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6773..6790)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7081..7100)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7311..7330)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7430..7452
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7490..7508)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7576..7680
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7681..7749,7750..8541)
                     /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(7899..7918)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8712..9300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9201..9220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9454..9471
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgatcccg ggaattctaa tacgactcac tatagctaat cgtcaggatg aagcggattc
       61 caaggaagac aaagggcaaa tccagcggaa agggcaatga ctcaacagag agagcggacg
      121 atggctcgag ccaattaaga gacaagcaaa acaataaggc tggccccgcc actacggagc
      181 ctggcacatc caaccgagag caatacaaag ctcgaccagg tattgcatct gtgcagaggg
      241 ccactgaaag tgcagaaatg cccatgaaga ataatgacga agggacgcca gataagaaag
      301 gaaatactaa gggcgaccta gttaatgagc atagtgaggc taaagacgag gcggatgaag
      361 cgacgaagaa gcaggcaaag gatacagaca aaagtaaagc gcaagtcaca tattcagaca
      421 ctggtatcaa taatgctaat gaactgtcaa gatctgggaa tgtggataat gagggtggaa
      481 gtaatcagaa gccgatgtct accagaatag ctgaggcaac gtctgctata gtgtcgaaac
      541 atcctgcgcg tgttgggctg ccacctaccg ctagcagtgg tcatgggtat cagtgccatg
      601 tctgttctgc agtcctgttt agtcctttag acctagatgc ccacgtcgcc tcacatggtt
      661 tgcatggtaa catgacatta acatcgagtg atatccagcg acatataact gagttcatca
      721 gctcatggca aaatcatcct attgttcaag tttcggctga tgtcgaaaat aagaaaactg
      781 ctcaattgct tcacgctgac actcctcgac tcgtcacttg ggatgctggt ttgtgtactt
      841 cattcaaaat cgtcccgatt gtgccagctc aggtgccgca ggatgtactg gcctatacgt
      901 ttttcacctc ttcatacgct atccaatcac cgtttccaga ggcggcagtg tctaggattg
      961 tggtgcatac gagatgggca tctaatgttg actttgaccg agactcgtct gtcatcatgg
     1021 cgccacctac agaaaacaat atccatttgt ttaaacagtt actaaatact gaaaccctgt
     1081 ctgtaagggg ggctaatccg ctaatgttca gggcgaatgt gttgcatatg ttgctagagt
     1141 tcgtattaga taacttgtat ctgaacagac atacgggatt ctctcaagac cacacgccat
     1201 ttactgaggg tgctaatttg cgttcacttc ctggccccga tgctgagaaa tggtactcga
     1261 ttatgtatcc aacgcgcatg ggaacgccga atgtatccaa aatatgtaat ttcgtcgcct
     1321 cttgtgtgcg aaatcgggtt ggacggtttg atcgagcaca gatgatgaac ggagctatgt
     1381 cagagtgggt ggatgtcttc gagacttcag acgcgctaac cgtctccatt cgaggtcgat
     1441 ggatggctag actagctcgc atgaacataa atccaacaga gatcgaatgg gcattgactg
     1501 aatgtgcaca aggatatgtg actgtcacaa gtccttacgc tcctatcgta aatagattga
     1561 tgccctatcg tatctccaac gctgagcggc aaatatcaca gataatcagg atcatgaaca
     1621 ttggcaataa cgcgacggtg atacaacctg ttctgcaaga tatttcggta ctccttcaac
     1681 gcatatcacc actccaaata gatccaacta ttatttccaa cactatgtca acagtctcgg
     1741 agtctactac tcagaccctc agccccgcgt cctcaatttt gggtaaacta cgaccaagca
     1801 actcagattt ttctagtttt agagtcgcgt tggctggatg gctttataat ggggttgtga
     1861 cgacggtgat tgatgatagt tcatatccaa aagacggcgg cagcgtgacc tcacttgaaa
     1921 atctgtggga tttcttcatc cttgcgcttg ctctaccact gacaactgac ccctgtgcac
     1981 ctgtgaaagc attcatgacc ctagccaaca tgatggttgg tttcgagaca atccctatgg
     2041 ataatcagat ctatactcaa tcgagacgcg cgagtgcttt ctcaacgcct cacacgtggc
     2101 cacgatgctt tatgaacatc cagttaattt ctccaatcga cgctcccatc ttgcgacagt
     2161 gggctgaaat tattcataga tactggccta acccttcaca gattcgttat ggtgcaccga
     2221 acgttttcgg ctcggcaaat ttgttcactc cacctgaggt gctgttattg ccaatcgatc
     2281 atcaaccagc taatgtaaca acgccaacgc tggacttcac caatgagtta actaattggc
     2341 gcgctcgtgt ctgtgagctt atgaagaatc tcgttgataa tcaaagatat caacctggat
     2401 ggacacaaag tctagtctcg tcaatgcgcg gaacgctaga caaattgaag ttgattaaat
     2461 cgatgacacc aatgtatctg caacagctgg ctccggtaga gttagcagtg atagctccca
     2521 tgttgccttt tccacctttc caggtgccat acgtccgtct cgatcgtgac agagttccaa
     2581 caatggttgg agtaacacga cagtcacgag atactattac tcagccggcg ctatcgctgt
     2641 cgacaaccaa tactactgtt ggcgtgccac tagctctaga cgcgagggct atcaccgttg
     2701 cgctgttgtc agggaaatat ccgccggatt tggtgacaaa tgtatggtac gctgatgcca
     2761 tttacccaat gtatgcagac acggaggtgt tctctaatct tcagagagac atgattacct
     2821 gcgaggccgt gcagacatta gtgactctgg tggcgcaaat atcagagacc cagtatcctg
     2881 tagataggta tcttgattgg atcccatcac tgagagcatc ggcggcgacg gcggcgacat
     2941 ttgctgagtg ggttaatact tcaatgaaga cggcgtttga tttgtctgat atgctgttag
     3001 agcctctcct aagcggtgat ccgaggatga ctcaactagc gattcagtat cagcagtaca
     3061 atggcagaac gtttaatatc atacctgaaa tgccaggttc agtaattgct gactgcgttc
     3121 aattaacagc agaagtcttt aatcacgaat ataacctgtt tgggattgcg cggggtgata
     3181 tcatcattgg ccgtgttcag tcgacacatt tgtggtcacc gctggctcct ccacctgacc
     3241 tggtgtttga tcgtgatacc cctggtgttc acatcttcgg acgagattgc cgtatatcgt
     3301 ttggaatgaa tggcgccgcg ccaatgatta gagatgagac tggactgatg gtgccttttg
     3361 aaggaaattg gattttccca ctggcgcttt ggcaaatgaa tacacgatat tttaatcaac
     3421 agttcgacgc gtggattaag acaggagagt tgcgaatccg cattgagatg ggcgcgtatc
     3481 catatatgtt gcattactat gatccacgtc agtacgctaa tgcatggaat ttaacatccg
     3541 cctggcttga agaaattacg ccgacgagca tcccatccgt gcctttcatg gtgcccattt
     3601 caagtgatca tgacatttcc tctgccccag ctgtccaata tatcatttca actgaatata
     3661 atgatcggtc tctgttctgc actaactcat catctcccca aaccatcgct ggaccagaca
     3721 aacacattcc agttgagaga tataacattc tgaccaaccc cgacgctcca cccacgcaga
     3781 tacaactgcc tgaagtcgtt gacttgtaca acgtcgtcac acgctatgcg tatgagactc
     3841 cgcctattac cgctgttgtt atgggtgttc cttgatcctc atcctcccaa caggtgctag
     3901 agcattgcgc tcaatgctag ttgggccgat tcatcgggtc ggcatggcat ctccacctcc
     3961 tcgcggtccg acctgggcat ccgaaggagg acgtcgtcca ctcggatggc taagggagag
     4021 ctcaaaaaaa aggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     4081 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     4141 ttgctgaaag gaggaactat atccggatcg agatcctcta gagtcgacct gcaggcatgc
     4201 aagctttaat acgactcact atagctattc gcggtcatgg cttacatcgc agttcctgcg
     4261 gtggtggatt cacgttcgag tgaggctatt ggactgctag aatcgtttgg agtagacgct
     4321 ggggctgacg cgaatgacgt ttcatatcaa gatcatgact atgtgttgga tcagttacag
     4381 tacatgttag atggatatga ggctggtgac gttatcgatg cactcgtcca caagaattgg
     4441 ttacatcact ctgtctattg cttgttgcca cccaaaagtc aactattaga gtattggaaa
     4501 agtaatcctt cagcgatacc ggacaacgtt gatcgtcggc ttcgtaaacg actaatgcta
     4561 aagaaagatc tcaggaaaga tgatgaatac aatcagctag cgcgtgcttt caagatatcg
     4621 gatgtctacg cacctctcat ctcatccacg acgtcaccga tgacaatgat acagaacttg
     4681 aatcaaggcg agatcgtgta caccacgacg gacagggtaa taggggctag aatcttgtta
     4741 tatgctccta gaaagtacta tgcgtcaact ctgtcattta ctatgactaa gtgcatcatt
     4801 ccgtttggta aagaggtggg tcgtgttcct cactctcgat ttaatgttgg cacattttcg
     4861 tcaattgcta ccccgaaatg ttttgtcatg agtggggttg atattgagtc catcccaaat
     4921 gaatttatca agttgtttta ccagcgcgtc aagagtgttc acgctaacat actaaatgac
     4981 atatctcctc agatcgtctc tgacatgata aacagaaagc gtctgcgcgt tcatactcca
     5041 tcagatcgtc gagccgcgca gttgatgcat ttgccttacc atgttaaacg aggagcgtct
     5101 cacgtcgacg tttacaaggt ggatgttgta gacatgttgt tcgaggtagt ggatgtggcc
     5161 gatgggttgc gcaacgtatc taggaaacta actatgcata ccgttccggt atgtattctt
     5221 gaaatgttgg gtattgagat tgcggactat tgcattcgtc gagaggatgg aatgctcaca
     5281 gattggttcc tacttttaac catgctatct gatggcttga ctgatagaag gacgcattgt
     5341 caatacttga ttaatccgtc aagtgtgcct cctgatgtga tacttaacat ctcaattact
     5401 ggatttataa atagacatac aatcgatgtc atgcctgaca tatatgactt tgttaaaccc
     5461 attggcgctg tgctgcctaa gggatcattt aaatcaacaa ttatgagagt tcttgattca
     5521 atatcaatat taggaatcca aatcatgccg cgcgcgcatg tagttgactc agatgaggtg
     5581 ggcgagcaaa tggagcctac gtttgagcag gcggttatgg agatatacaa agggattgct
     5641 ggcgttgact cgctggatga tctcatcaag tgggtgctga actcggatct cattccgcat
     5701 gatgacaggc ttggtcaatt atttcaagcg tttttgcctc tcgcaaagga cttattagct
     5761 ccaatggcca gaaagtttta tgataactca atgagtgagg gtagattgct aacattcgct
     5821 catgccgaca gtgagttgct gaacgcaaat tattttggtc atttattgcg actaaaaata
     5881 ccatatatta cagaggttaa tctgatgatt cgcaagaatc gtgagggtgg agagctattt
     5941 cagcttgtgt tatcttatct atataaaatg tatgctacta gcgcgcagcc taaatggttt
     6001 ggatcattat tgcgattgtt aatatgtccc tggttacata tggagaaatt aataggagaa
     6061 gcagacccgg catctacgtc ggctgaaatt gggtggcata tccctcgtga acagctgatg
     6121 caagatggat ggtgtggatg tgaagacgga ttcattccct atgttagcat acgtgcgcca
     6181 agactggtta tagaggagtt gatggagaag aactggggcc aatatcatgc ccaagttatt
     6241 gtcactgatc agcttgtcgt aggcgaaccg cggagggtat ctgctaaggc tgtgatcaag
     6301 ggtaaccact taccagttaa gttagtttca cgatttgcat gtttcacatt gacggcgaag
     6361 tatgagatga ggctttcgtg cggccatagc actggacgtg gagctgcata cagtgcgaga
     6421 ctagctttcc gatctgactt ggcgtgatcc gtgacatgcg tagtgtgaca cctgctccta
     6481 ggtcaatggg ggtagggggc gggctaagac tacgtacgcg cttcatcggg tcggcatggc
     6541 atctccacct cctcgcggtc cgacctgggc atccgaagga ggacgtcgtc cactcggatg
     6601 gctaagggag agctcaaaaa aaaggatccg gctgctaaca aagcccgaaa ggaagctgag
     6661 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     6721 ttgaggggtt ttttgctgaa aggaggaact atatccggat caagcttgta ttctatagtg
     6781 tcacctaaat cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa
     6841 cgacaatatg tacaagccta attgtgtagc atctggctta ctgaagcaga ccctatcatc
     6901 tctctcgtaa actgccgtca gagtcggttt ggttggacga accttctgag tttctggtaa
     6961 cgccgttccg caccccggaa atggtcagcg aaccaatcag cagggtcatc gctagccaga
     7021 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
     7081 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
     7141 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
     7201 ccttgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     7261 cctaatgcag gagtcgcata agggagagcg tcgatatggt gcactctcag tacaatctgc
     7321 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     7381 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     7441 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     7501 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     7561 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     7621 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     7681 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     7741 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     7801 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     7861 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     7921 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7981 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     8041 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     8101 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     8161 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     8221 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     8281 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     8341 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     8401 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     8461 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     8521 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     8581 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     8641 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     8701 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     8761 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     8821 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     8881 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     8941 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     9001 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     9061 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     9121 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     9181 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     9241 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     9301 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     9361 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     9421 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     9481 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcaggggg
     9541 at
//
