LOCUS       sequence-81251-e        7164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression (Tet-Off system) of the rainbow trout antiviral
            Mx1 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7164)
  AUTHORS   Lester K, Hall M, Urquhart K, Gahlawat S, Collet B
  TITLE     Development of an in vitro system to measure the sensitivity to the 
            antiviral Mx protein of fish viruses.
  JOURNAL   J Virol Methods. 2012 Jun;182(1-2):1-8. doi: 
            10.1016/j.jviromet.2012.01.014. Epub 2012 Mar 1.
  PUBMED    22405879
REFERENCE   2  (bases 1 to 7164)
  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: "J Virol 
            Methods."; date: "2012-06"; volume: "182(1-2):1-8. doi"; pages: " 
            10.1016/j.jviromet.2012.01.014. Epub 2012 Mar 1"
COMMENT     Sequence Label: pTRE2-Hyg-omMx1
FEATURES             Location/Qualifiers
     source          1..7164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(15..33,34..56,57..75,76..98,99..117,118..140,141..159,
                     160..182,183..201,202..224,225..243,244..266,267..285)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        318..356
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     353..377
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          2393..2965
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3097..3116)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3162..3217
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3216..3235)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3608..3625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3779..4367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3859..3878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4538..5329,5330..5398))
                     /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     5161..5180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5399..5503)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5571..5589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    5604..5685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5629..5648
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5802..6827)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(5802..6827)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(join(6836..7164,1))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6850..6985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ctcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
       61 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
      121 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
      181 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      241 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
      301 ctcggtaccc gggtcgaggt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt
      361 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
      421 accgggaccg atccagcctc cgcggccccg aattcgagct cggtacccgg ggatcctcta
      481 gtcagctgac gcgtatgaat aatacgctca accaacatta cgaagagaag gtgcggccct
      541 gtatcgacct catcgactcc ctgcgctccc ttggcgtaga gaaggatctt gcgctgccag
      601 ccatcgccgt gataggggac cagagttcgg ggaagagctc cgtgttggag gcgctgtctg
      661 gggtggcttt gccaaggggt agcggtattg taacacgatg ccctctcgag ctgaagatga
      721 aaaggaagaa agaaggagag gaatggcacg gaaaaatcag ctaccaagac catgaggagg
      781 agattgagga tccctctgat gtggagaaga aaattcgtga agcccaggat gaaatggcag
      841 gtgtgggggt gggtatcagt gatgacctca tcagcctaga gattggctcc cctgacgtcc
      901 cagacctcac actcatcgac ctgccaggca tcgcccgggt agctgtcaag ggtcaacctg
      961 agaacattgg tgaacagatt aagagactga tacggaaatt catcatgaag caagaaacaa
     1021 tcagcttggt ggttgtgcca tgcaacgttg acattgcaac cacagaggct ttgaagatgg
     1081 cacaagaggt ggaccctgaa ggggaaagga cattaggcat cctgaccaag cctgacctgg
     1141 tagacaaagg cacagaggag acggtggtgg acatagttca taatgaggtt atccacctga
     1201 ctaagggcta catgatagtc aagtgcaggg gccagaagga aatcatggag cgagtctcac
     1261 tgaccgaggc cacagagagg gagaaggctt tcttcaaaga gcacgctcat ctcagcacac
     1321 tttatgatga gggccatgcc accatcccta aactggcaga gaaattaact cttgaattgg
     1381 tgcatcatat cgagaaatcc ctgcctcgtc tagaagagca gattgaggca aagctgtcag
     1441 agacacatgc cgagctggaa agatatggta ccggaccccc tgaggactcg gcagagaggc
     1501 tgtatttcct gatcgataaa gtgactgcat tcacccaaga tgccatcaac ctgagcactg
     1561 gggaggagat gaaaagcgga gttcgtctca acgtcttctc cacactcaga aaagagtttg
     1621 ggaaatggaa gttacacctg gaacgctctg gagaaatctt taaccagagg attgagggag
     1681 aagtggatga ttatgagaag acgtaccgtg gaagggagct cccagggttc atcaactaca
     1741 agacatttga ggtgatggtg aaagaccaga tcaaacaact ggagggacca gcagtcaaga
     1801 aactgaagga gatttcagat gccgttagga aggtgttctt actgctggct cagagcagct
     1861 tcactggatt tcctaacctc ctgaaatcag cgaagacaaa gattgaggcc attaagcagg
     1921 tgaatgagtc tacagctgag tccatgttga ggactcagtt caagatggag ctgatagtgt
     1981 acacacagga cagcacctac agccacagtc tgtgtgagag gaagagggag gaggacgaag
     2041 accaaccctt aactgagata aggagtacga tctttagcac agacaaccat gccaccctac
     2101 aggagatgat gctgcacctc aagtcctact actggatatc cagtcagcgt ctggctgatc
     2161 agattcccat ggtgatccgc tacctggtgc tgcaggagtt tgcttcccag ctgcagaggg
     2221 agatgcttca gactctgcag gagaaggaca acatcgagca gctgctgaag gaggacatcg
     2281 acatcggcag caaaagggct gcactgcaga gcaagctcaa acgcctgatg aaggcacgca
     2341 gctacctagt tgagttctag tcgacgatat ctctagagct gagaacttca gggtgagttt
     2401 ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat atggaggggg
     2461 caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc atggaccctc
     2521 atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc ctcttatttt
     2581 cttttcattt tctgtaactt ttttcgttaa actttagctt gcatttgtaa cgaattttta
     2641 aattcacttt cgtttatttg tcagattgta agtactttct ctaatcactt ttttttcaag
     2701 gcaatcaggg taattatatt gtacttcagc acagttttag agaacaattg ttataattaa
     2761 atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata ttcttattgg
     2821 tagaaacaac tacatcctgg taatcatcct gcctttctct ttatggttac aatgatatac
     2881 actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg ctaaccatgt
     2941 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca
     3001 tcattttggc aaagaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     3061 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     3121 tggggacatc atgaagcccc ttgagcatct gacttctggg taataaagga aatttatttt
     3181 cattgcaata gtgtgtggga attttttgtg tctctcactc ggaaggacat atgggagggc
     3241 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgccatatg
     3301 ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa cagccccctg
     3361 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     3421 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     3481 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatgaactc
     3541 gactgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3601 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3661 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3721 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3781 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3961 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4201 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4381 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4441 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4501 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4561 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4621 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4681 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4741 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4801 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4861 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4921 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4981 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5041 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5101 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5161 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5221 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5281 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5341 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5401 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5461 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5521 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5581 tcacgaggcc ctttcgtctc gaggcagtga aaaaaatgct ttatttgtga aatttgtgat
     5641 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     5701 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     5761 ctctacaaat gtggtatggc tgattatgat cctctagaac tctattcctt tgccctcgga
     5821 cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag
     5881 acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg
     5941 attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc
     6001 tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca
     6061 agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc
     6121 ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag
     6181 tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg
     6241 tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc
     6301 tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga
     6361 tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg
     6421 aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catggcctcc
     6481 gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc
     6541 tgtgcacggc gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact
     6601 tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa
     6661 ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa
     6721 gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg
     6781 atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatgga agctttttgc
     6841 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     6901 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     6961 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     7021 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     7081 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     7141 actttccaca ccctaactga caca
//
