LOCUS       sequence-195316-        6137 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 6137)
  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 6137)
  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..6137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..28)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    join(68..86,87..109,110..128,129..151,152..170,171..193,
                     194..212,213..235,236..254,255..277,278..296,297..319,
                     320..338)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        371..409
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     406..430
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             595..621
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1834..1857
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     intron          1888..2460
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2592..2611)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2657..2712
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2711..2730)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3110..3127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3281..3869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3361..3380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4040..4831,4832..4900))
                     /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     4663..4682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4901..5005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5073..5091
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5110..5439
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5290..5425
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5352..5371
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5487..5534
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5553..5951
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    join(6109..6137,1..53)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcctcgagt
       61 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
      121 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
      181 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct
      241 atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag
      301 tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagctcggta
      361 cccgggtcga ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa
      421 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
      481 ccgatccagc ctccgcggcc ccgaattcga gctcggtacc cggggatcct ctagtcagct
      541 gacgcgtgct agcgcggccg catcgataag cttgtcgacg aattcatgac tagttaccca
      601 tacgatgttc cagattacgc tactagtgag cccaggaggg cggcgcccgg ggtgcctggc
      661 tgggggtctc gggaggccgc ggggtcggca tcggccgcgg agcttgtgta ccatctagcc
      721 ggggccctgg gcactgagct gcaggatctg gcgcgccgtt tcgggccgga ggcggcggcc
      781 gggctggtgc cgctagtggt gcgggcgctg gagctcttgg aacaggctgc cgtggggccc
      841 gccccggact cgctgcaggt gtcggcgcag ccggcggagc aggagctgcg gcggctgcgg
      901 gaggagaacg agcgcctccg cagggagctg cgcgcggggc cacaggagga gcgcgcgctg
      961 ctgcggcagc tcaaggaggt cacggaccga cagcgggacg aactccgggc gcacaaccgc
     1021 gacctgcggc agcgcggcca ggagaccgag gcgttgcagg agcagctgca gcgcctcctg
     1081 ctggtgaacg ctgagctgcg gcacaagctg gcggccatgc agacccagct gcgcgccgcg
     1141 caggaccgcg agagggagcg ccagcagcct ggcgaagccg cgactccgca ggctaaagag
     1201 cgagcgcggg ggcaggccgg gcggcccggg caccagcacg gacaggagcc cgaatgggcg
     1261 accgccggcg caggcgcccc agggaaccct gaggacccgg cggaggccgc gcagcagctc
     1321 gggcgcccct cggaggcagg gcagtgccgc ttcagtcggg aggagtttga gcagatcctt
     1381 caggagcgga atgaactcaa agccaaagtg ttcctgctca aggaggaact ggcctacttc
     1441 cagcgggagc tgctcacaga ccaccgggtc cccggccttc tgctcgaggc catgaaggtg
     1501 gctgtccgga agcagcggaa gaagatcaag gccaagatgt tagggacacc agaggaagca
     1561 gagagcagtg aggatgaggc tggcccatgg atcctgctct ccgatgacaa gggagaccat
     1621 cccccacccc cggagtccaa aatacagagt ttctttggcc tatggtatcg gggtaaagct
     1681 gaatcctctg aggatgagac cagcagccct gcacccagca agctaggggg agaagaggag
     1741 gcccaaccac agtctccagc tcctgatccg ccctgttctg ccctccacga acacctttgt
     1801 ctgggggcct cagccgcccc agaggccacg cgtgattata aagatgatga tgataaaacg
     1861 cgttgatcta gagctgagaa cttcagggtg agtttgggga cccttgattg ttctttcttt
     1921 ttcgctattg taaaattcat gttatatgga gggggcaaag ttttcagggt gttgtttaga
     1981 atgggaagat gtcccttgta tcaccatgga ccctcatgat aattttgttt ctttcacttt
     2041 ctactctgtt gacaaccatt gtctcctctt attttctttt cattttctgt aactttttcg
     2101 ttaaacttta gcttgcattt gtaacgaatt tttaaattca cttttgttta tttgtcagat
     2161 tgtaagtact ttctctaatc actttttttt caaggcaatc agggtatatt atattgtact
     2221 tcagcacagt tttagagaac aattgttata attaaatgat aaggtagaat atttctgcat
     2281 ataaattctg gctggcgtgg aaatattctt attggtagaa acaactacac cctggtcatc
     2341 atcctgcctt tctctttatg gttacaatga tatacactgt ttgagatgag gataaaatac
     2401 tctgagtcca aaccgggccc ctctgctaac catgttcatg ccttcttctc tttcctacag
     2461 ctcctgggca acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcactcct
     2521 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     2581 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     2641 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     2701 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     2761 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     2821 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     2881 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     2941 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     3001 ctcccagtca tagctgtccc tcttctctta tggagatccc tcgactgcat taatgaatcg
     3061 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3181 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3361 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3721 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4621 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     4681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5041 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5101 ttcactcgag tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5161 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5221 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5281 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5341 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     5401 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     5461 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     5521 tagtataata cgacaaggtg aggaactaaa ccatggccaa gcctttgtct caagaagaat
     5581 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     5641 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     5701 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     5761 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     5821 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     5881 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     5941 gggagggcta agcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     6001 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     6061 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     6121 gcagcttata atggtta
//
