LOCUS       sequence-193578-        6139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual tagged RILP for expression in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6139)
  AUTHORS   Wozniak AL, Long A, Jones-Jamtgaard KN, Weinman SA
  TITLE     Hepatitis C virus promotes virion secretion through cleavage of the 
            Rab7 adaptor protein RILP.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Nov 1;113(44):12484-12489. Epub 2016 
            Oct 17.
  PUBMED    27791088
REFERENCE   2  (bases 1 to 6139)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2016-11-1"; volume: "113(44)"; pages: 
            "12484-12489. Epub 2016 Oct 17"
COMMENT     Sequence Label: pTRE2-Bla(HA–RILP-FLAG)
FEATURES             Location/Qualifiers
     source          1..6139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..335
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      186..321
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     248..267
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        383..430
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             449..847
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    1005..1086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1042..1061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    join(1101..1119,1120..1142,1143..1161,1162..1184,1185..1203,
                     1204..1226,1227..1245,1246..1268,1269..1287,1288..1310,
                     1311..1329,1330..1352,1353..1371)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1404..1442
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1439..1463
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1628..1654
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2867..2890
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     intron          2921..3493
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3625..3644)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3690..3745
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3744..3763)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4143..4160)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4314..4902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4394..4413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5073..5864,5865..5933))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5696..5715
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5934..6038)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6106..6124
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
       61 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
      121 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
      181 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
      241 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
      301 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
      361 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
      421 ataatacgac aaggtgagga actaaaccat ggccaagcct ttgtctcaag aagaatccac
      481 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
      541 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
      601 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
      661 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
      721 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
      781 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
      841 gggctaagca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
      901 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
      961 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     1021 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1081 cactgcctcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
     1141 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     1201 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     1261 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
     1321 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     1381 gtcgagctcg gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag
     1441 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1501 gaagacaccg ggaccgatcc agcctccgcg gccccgaatt cgagctcggt acccggggat
     1561 cctctagtca gctgacgcgt gctagcgcgg ccgcatcgat aagcttgtcg acgaattcat
     1621 gactagttac ccatacgatg ttccagatta cgctactagt gagcccagga gggcggcgcc
     1681 cggggtgcct ggctgggggt ctcgggaggc cgcggggtcg gcatcggccg cggagcttgt
     1741 gtaccatcta gccggggccc tgggcactga gctgcaggat ctggcgcgcc gtttcgggcc
     1801 ggaggcggcg gccgggctgg tgccgctagt ggtgcgggcg ctggagctct tggaacaggc
     1861 tgccgtgggg cccgccccgg actcgctgca ggtgtcggcg cagccggcgg agcaggagct
     1921 gcggcggctg cgggaggaga acgagcgcct ccgcagggag ctgcgcgcgg ggccacagga
     1981 ggagcgcgcg ctgctgcggc agctcaagga ggtcacggac cgacagcggg acgaactccg
     2041 ggcgcacaac cgcgacctgc ggcagcgcgg ccaggagacc gaggcgttgc aggagcagct
     2101 gcagcgcctc ctgctggtga acgctgagct gcggcacaag ctggcggcca tgcagaccca
     2161 gctgcgcgcc gcgcaggacc gcgagaggga gcgccagcag cctggcgaag ccgcgactcc
     2221 gcaggctaaa gagcgagcgc gggggcaggc cgggcggccc gggcaccagc acggacagga
     2281 gcccgaatgg gcgaccgccg gcgcaggcgc cccagggaac cctgaggacc cggcggaggc
     2341 cgcgcagcag ctcgggcgcc cctcggaggc agggcagtgc cgcttcagtc gggaggagtt
     2401 tgagcagatc cttcaggagc ggaatgaact caaagccaaa gtgttcctgc tcaaggagga
     2461 actggcctac ttccagcggg agctgctcac agaccaccgg gtccccggcc ttctgctcga
     2521 ggccatgaag gtggctgtcc ggaagcagcg gaagaagatc aaggccaaga tgttagggac
     2581 accagaggaa gcagagagca gtgaggatga ggctggccca tggatcctgc tctccgatga
     2641 caagggagac catcccccac ccccggagtc caaaatacag agtttctttg gcctatggta
     2701 tcggggtaaa gctgaatcct ctgaggatga gaccagcagc cctgcaccca gcaagctagg
     2761 gggagaagag gaggcccaac cacagtctcc agctcctgat ccgccctgtt ctgccctcca
     2821 cgaacacctt tgtctggggg cctcagccgc cccagaggcc acgcgtgatt ataaagatga
     2881 tgatgataaa acgcgttgat ctagagctga gaacttcagg gtgagtttgg ggacccttga
     2941 ttgttctttc tttttcgcta ttgtaaaatt catgttatat ggagggggca aagttttcag
     3001 ggtgttgttt agaatgggaa gatgtccctt gtatcaccat ggaccctcat gataattttg
     3061 tttctttcac tttctactct gttgacaacc attgtctcct cttattttct tttcattttc
     3121 tgtaactttt tcgttaaact ttagcttgca tttgtaacga atttttaaat tcacttttgt
     3181 ttatttgtca gattgtaagt actttctcta atcacttttt tttcaaggca atcagggtat
     3241 attatattgt acttcagcac agttttagag aacaattgtt ataattaaat gataaggtag
     3301 aatatttctg catataaatt ctggctggcg tggaaatatt cttattggta gaaacaacta
     3361 caccctggtc atcatcctgc ctttctcttt atggttacaa tgatatacac tgtttgagat
     3421 gaggataaaa tactctgagt ccaaaccggg cccctctgct aaccatgttc atgccttctt
     3481 ctctttccta cagctcctgg gcaacgtgct ggttgttgtg ctgtctcatc attttggcaa
     3541 agaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     3601 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     3661 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     3721 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     3781 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     3841 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     3901 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     3961 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     4021 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgactg
     4081 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4141 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4201 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4261 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4321 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4381 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4441 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4501 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4561 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4621 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4681 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4741 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4801 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4861 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4921 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4981 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5041 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5101 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5161 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5221 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5281 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5341 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5401 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5461 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5521 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5581 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5641 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     5701 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5761 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5821 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5881 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5941 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6001 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6061 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     6121 aggccctttc gtcttcaca
//
