LOCUS       sequence-39537-e        9494 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 9494)
  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 9494)
  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-L2-M3T3D
FEATURES             Location/Qualifiers
     source          1..9494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             779..787
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      4111..4158
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(4226..4244)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4227..4244)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      6889..6936
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(7033..7052)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7263..7282)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7382..7404
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7442..7460)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7528..7632
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7633..7701,7702..8493)
                     /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(7851..7870)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8664..9252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9153..9172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9406..9423
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgatcccg ggaattctaa tacgactcac tatagctaaa aggcgcgatg gcgaacgttt
       61 ggggggtgag acttgcagac tcgttatctt cacccactat tgagacacga acgcgtcagt
      121 ataccttaca cgatctttgc tcagacctag atgctaatcc ggggagggaa ccgtggaaac
      181 ctctgcgtaa tcagcgtact aataatattg tggctgtgca attattcaga ccattgcagg
      241 gtttagtttt agatacccag ctttatggat ttccaggagc atttgatgac tgggagcgat
      301 tcatgagaga gaagctgcgt gtgctaaagt atgaagtatt gcgcatctat ccaatcagca
      361 actatagcaa tgaacatgtc aacgtcttcg tggccaatgc tttggtgggc gctttcctgt
      421 cgaatcaagc tttctatgac ctgctaccgt tgttgataat taatgacact atgattggtg
      481 atctacttgg cacgggggca tcgctatcac agttctttca atctcatgga gatgtgctgg
      541 aagtcgcagc tggtcgtaag tatctgcaga tggaaaacta ctccaacgat gacgatgatc
      601 ctccattatt tgcgaaagac ctgtcagatt atgctaaagc attctacagt gacacatatg
      661 aagtgttgga caggttcttt tggacgcatg actcttcagc gggggtctta gtgcattatg
      721 ataagccaac gaatggtcat cactatctgc tgggtacttt gactcagatg gtcagtgcac
      781 ctccttatat tattaacgct actgacgcaa tgttgcttga atcctgtcta gaacagttct
      841 cagctaatgt gcgtgcgaga cctgcgcaac ccgttacacg cttagaccaa tgctatcatt
      901 taagatgggg agcacaatat gtaggagaag attcactgac atatcggttg ggggtgttat
      961 ccttgctggc taccaatgga tatcaattag ctagaccgat tccaagacag ttgacgaatc
     1021 gatggttgtc gagctttgtg agtcaaatta tgtctgacgg cgtcaacgag actccactgt
     1081 ggccccaaga aaggtatgtg cagatcgctt atgattcacc atccgttgtt gatggggcta
     1141 cgcaatatgg ctatgtcagg aagaatcaac tcagactcgg catgagaata tcggcgctgc
     1201 aatcgctgag tgatacgccc tcgccggtac agtggcttcc acaatacacc atcgaccagg
     1261 cagcgatgga cgaaggcgat ctgatggtta gtcggcttac gcaactcccg ttacgtcctg
     1321 attatggtaa tatctgggtc ggcgatgcgc tatcctatta tgtggactac aatcggagtc
     1381 atcgagtcgt gctttcatcg gaacttcctc agcttccgga cacatatttt gatggcgatg
     1441 aacagtatgg gcgcagcctg ttctcactag ctcgtaagat tggtgaccgc tcgttagtga
     1501 aagatacggc tgtcttgaag cacgcttacc aagccatcga tccaaatact ggtaaggggt
     1561 atctgagatc tgggcaatct gtcgcatatt ttggtgcatc agcgggtcat tctggtgccg
     1621 accagccgtt agtcatagag ccctggattc aagggaaaat cagtggtgtg ccgccaccct
     1681 cctcagtgcg acagttcggc tatgatgttg cccgtggcgc gatcgtcgat ctggcgagac
     1741 catttccttc tggagattat caatttgtct attcggatgt tgaccaggtg gtcgatggcc
     1801 atgacgatct gagtatatca tctggactgg tggagagcct tttgtcttca tgcatgcacg
     1861 ccacagcacc cgggggctca tttgttgtta agataaattt tccgactaga cccgtatggc
     1921 actacatcga acagaagatc ttgcccaata ttacgtcata catgttgatc aagcctttcg
     1981 tcaccaacaa cgtcgaattg ttcttcgtcg ctttcggtgt gcatcaacac tcatcactta
     2041 cttggacatc tggagtgtac ttcttcttgg tggaccattt ttatcgttat gagactttat
     2101 ctacgatctc acgacaattg ccgtcttttg ggtatgttga tgatgggtct tccgtgactg
     2161 gtatcgagac aattagtatt gagaaccctg gcttctcgaa tatgacccag gccgctcgca
     2221 ttggtatctc aggattgtgt gctaatgtag gtaacgcgcg taagtccatt gccatttacg
     2281 aatcccatgg ggccagagta ttaactatca catcaaggag atctccggca tcagctagaa
     2341 gaaagtctag gttgcgatat ttgccattaa tagaccctag gtcgttagag gtacaggcgc
     2401 gcactattct gccagctgat ccagtgttat ttgaaaacgt gagcggagcg tcaccccatg
     2461 tttgtctgac aatgatgtac aacttcgaag tgtcgtcagc ggtatatgat ggagacgttg
     2521 tgctagatct tgggacggga ccagaggcta aaatccttga actgataccc gcaacctctc
     2581 cagtcacatg cgtggacata cggcctacag cgcagcctag tggatgttgg aacgttcgta
     2641 ccacgttcct tgagttagat tatttgagcg atggatggat cactggggtg cgtggggaca
     2701 tagttacttg tatgttatct ttgggggccg ctgccgctgg aaaatcaatg acttttgacg
     2761 ctgcgtttca gcaattaatc aaagtattat ccaagagtac ggctaatgtt gtgctggtgc
     2821 aggttaactg ccctacagac gtggtgagga gcattaaggg ctacctagag atagattcga
     2881 ctaacaagag gtataggttc cccaaatttg gtcgagacga gccgtactct gacatggatg
     2941 cgctggagaa aatatgtcgt accgcctggc caaactgctc aattacctgg gttccattgt
     3001 catacgactt gcggtggact agactggcat tattagagtc cacgacattg agtagcgcgt
     3061 cgattagaat tgctgagctg atgtataaat acatgcctat tatgaggatt gacattcatg
     3121 gactacccat ggaaaagcga ggtaacttca tagtggggca gaactgctca ttagtaatcc
     3181 ctggttttaa tgcgcaggat gtctttaact gttatttcaa ttccgccctc gctttctcga
     3241 ctgaagatgt caatgctgcg atgattcccc aagtgtctgc gcagtttgat gcgactaagg
     3301 gtgagtggac gttggatatg gtcttctccg acgcaggaat ctataccatg caggctctag
     3361 tgggatctaa tgctaatcca gtctctttgg gttcctttgt agttgattct ccagatgtag
     3421 atataactga cgcttggcca gctcagttag actttacgat cgcgggaact gatgtcgata
     3481 taacagttaa tccttattac cgtctgatga cctttgtaag gatcgatgga cagtggcaga
     3541 ttgccaatcc agacaaattt caattctttt cgtcggcgtc tgggacgtta gtgatgaacg
     3601 tcaaattaga tatcgcagat aaatatctac tatactatat acgagatgtc cagtctcgag
     3661 atgttggctt ttacattcag catccacttc aacttttgaa tacgatcaca ttgccaacca
     3721 acgaggacct ttttctgagc gcacctgaca tgcgagagtg ggcagttaag gaaagcggta
     3781 acacgatatg tatactcaat agtcaagggt ttgtgctacc tcaagattgg gatgtgttaa
     3841 cagataccat aagttggtcc ccatcgatac ccacatacat tgtgccaccg ggtgattata
     3901 ccttgactcc tctgtaactc actgtccctc gtgagcgcgc ctaattcatc gggtcggcat
     3961 ggcatctcca cctcctcgcg gtccgacctg ggcatccgaa ggaggacgtc gtccactcgg
     4021 atggctaagg gagagctcaa aaaaaaggat ccggctgcta acaaagcccg aaaggaagct
     4081 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     4141 gtcttgaggg gttttttgct gaaaggagga actatatccg gatcgagatc ctctagagtc
     4201 gacctgcagg catgcaagct tgtattctat agtgtcacct aaatcgtatg tgtatgatac
     4261 ataaggttat gtattaattg tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg
     4321 tagcatctgg cttactgaag cagaccctat catctctctc gtaaactgcc gtcagagtcg
     4381 gtttggttgg acgaaccttc tgagtttctg gtaacgccgt tccgcacccc ggaaatggtc
     4441 agcgaaccaa tcagcagggt catcgctagc taatacgact cactatagct aaagtgaccg
     4501 tggtcatggc ttcattcaag ggattctccg ccaacactgt tccagtttct aaggccaagc
     4561 gtgacatatc atctcttgcc gctactcctg gacttcgttc acaatccttc actccgtctg
     4621 tggatatgtc tcaatcgcgt gaattcctca caaaggcaat tgagcaaggg tccatgtcta
     4681 taccttatca gcatgtgaat gtaccgaaag ttgatcgtaa agttgttagc ctggtagtgc
     4741 gacctttctc ttcaggtgct ttctctatct ctggagtgat ttcgccagcc catgcctatc
     4801 tactagagtg tctaccccag cttgagcagg cgatggcttt tgttgcttca cctgagtctt
     4861 tccaggcttc cgacgtcgcg aagcgctttg ccataaagcc aggtatgagc ctccaggatg
     4921 ccatcactgc ctttattaac tttgtgtccg cgatgctgaa aatgacggtg actcgtcaaa
     4981 actttgacgt tattgtggct gagatcgaga ggcttgcttc aaccagcgtg tccgtcagga
     5041 ctaaagaagc gaaggttgct gatgaggagc taatgctatt cgggttagat catagagggc
     5101 cacagcagct ggatgtttct gacgctaaag ggataatgaa ggctgctgat attcagacaa
     5161 ctcatgatgt ccatttggca ccaggcgttg gtaatattga tcctgaaatc tataacgagg
     5221 ggcggttcat gttcatgcag cacaagccac ttgcggcgga tcaatcgtat ttcaccttgg
     5281 agactgcgga ttatttcaag atttatccaa catacgatga acatgatggc aggatggctg
     5341 accaaaagca gtcgggattg atactgtgta ctaaggacga ggtattggct gagcaaacta
     5401 tatttaaact ggacgcccct gatgacaaga ctgttcatct gttggatcgc gatgacgacc
     5461 acgttgttgc cagatttact aaggtattta tagaggacgt ggctcccggg catcatgctg
     5521 ctcaaagatc gggacaacgc tctgtgcttg atgacctata tgcgaatacg caagtgattt
     5581 ccattacttc tgctgcttta aagtgggtgg tcaagcacgg cgtatctgat ggaatcgtga
     5641 acaggaagaa tgtcaaagtg tgtgttggtt ttgaccccct gtacaccttg tctacacata
     5701 acggggtgtc cttatgtgcc ctgctgatgg acgaaaaact ctctgtgctg aacagtgcgt
     5761 gtcgtatgac gttacgctca ctcatgaaga ccggacgcga cgttgatgca cacagagctt
     5821 ttcagcgagt cctctctcaa ggatacacat cgctaatgtg ctactatcat ccttcacgga
     5881 agttggcata tggtgaggtg ctctttctag aacgatccaa tgacgtgaca gatgggatca
     5941 agcttcagtt ggacgcatct agacagtgtc atgaatgtcc tgtgttgcag cagaaagtgg
     6001 ttgagttaga gaaacagatt attatgcaga agtcaatcca gtcagaccct accccagtgg
     6061 cgctgcaacc attgttgtct cagttgcgtg agttgtctag tgaagttact aggctacaga
     6121 tggagttgag tcgagctcag tccctgaatg ctcagttgga ggcggatgtc aagtcagctc
     6181 aatcatgtag cttggatatg tatctgagac accacacttg cattaatggt catgctaaag
     6241 aagatgaatt gcttgacgct gtgcgtgtcg cgccggatgt gaggagagaa atcatggaaa
     6301 agaggagtga agtgagacaa ggttggtgcg aacgtatttc taaggaagca gctgccaaat
     6361 gtcaaactgt tattgatgac ctgactttga tgaatggaaa gcaagcacaa gagataacag
     6421 aattacgtga ttcggctgaa aaatatgaga aacagattgc agagctggtg agtaccatca
     6481 cccaaaacca gataacgtat cagcaagagc tacaagcctt ggtagcgaaa aatgtggaat
     6541 tggacgcgtt gaatcagcgt caggctaagt ctttgcgtat tactccctct cttctatcag
     6601 ccactcctat cgattcagct gatggtgttg ctgacttaat tgatttctct gttccaactg
     6661 atgagttgta aataatccgt gatgcagtgt tgccctaatc ccttaagcct tcccgacccc
     6721 cattcatcgg gtcggcatgg catctccacc tcctcgcggt ccgacctggg catccgaagg
     6781 aggacgtcgt ccactcggat ggctaaggga gagctcaaaa aaaaggatcc ggctgctaac
     6841 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     6901 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     6961 tcgctagcca gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
     7021 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
     7081 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
     7141 tgggcgccat ctccttgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     7201 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgatatg gtgcactctc
     7261 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     7321 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     7381 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     7441 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     7501 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     7561 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     7621 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     7681 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     7741 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     7801 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     7861 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     7921 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     7981 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     8041 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     8101 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     8161 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     8221 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     8281 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     8341 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     8401 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     8461 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     8521 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     8581 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     8641 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     8701 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     8761 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     8821 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     8881 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     8941 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     9001 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     9061 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     9121 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     9181 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     9241 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     9301 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     9361 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     9421 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     9481 taatgcaggg ggat
//
