LOCUS       sequence-318393-        8717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses soluble IL6 receptor in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8717)
  AUTHORS   Khatib A, Solaimuthu B, Ben Yosef M, Abu Rmaileh A, Tanna M, Oren G,
            Schlesinger Frisch M, Axelrod JH, Lichtenstein M, Shaul YD
  TITLE     The glutathione peroxidase 8 (GPX8)/IL-6/STAT3 axis is essential in 
            maintaining an aggressive breast cancer phenotype.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Sep 1;117(35):21420-21431. doi: 
            10.1073/pnas.2010275117. Epub 2020 Aug 18.
  PUBMED    32817494
REFERENCE   2  (bases 1 to 8717)
  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"; doi: 
            "10.1073/pnas.2010275117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-09-1- 1"; volume: "117"; issue: "35"; pages: 
            "21420-21431"
COMMENT     Sequence Label: sIL6R PLAK1
FEATURES             Location/Qualifiers
     source          1..8717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             185..229
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     primer_bind     527..547
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        836..1039
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     990..1010
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2177..2242
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2290..2407
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2456..2966
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2885..2903
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     LTR             3930..4163
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4235..4356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4272..4291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4326..4345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4396..4531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4551..4570)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4552..4570)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4580..4597)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4580..4596)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4589..4611)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4738..5193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4825..4844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5035..5056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5219..5323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5324..5392,5393..6184)
                     /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(5542..5561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6355..6943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6844..6863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7097..7114
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7231..7252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7267..7272,7273..7290,7291..7297)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7305..7321
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7310..7332
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7329..7345
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7329..7345
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7364..7384
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7366..7384
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7412..7638
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7639..7819
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7866..7991
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8484..8717
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
       61 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
      121 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
      181 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
      241 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
      301 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
      361 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
      421 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg
      481 acggtatcga tcacgagact agcctcgagc agcagccccc ttcaccgagg gcctatttcc
      541 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa
      601 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
      661 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc caagtacgcc
      721 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      781 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      841 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      901 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      961 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     1021 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtttaa
     1081 ttaatccaac atgctggccg tcggctgcgc gctgctggct gccctgctgg ccgcgccggg
     1141 agcggcgctg gccccaaggc gctgccctgc gcaggaggtg gcgagaggcg tgctgaccag
     1201 tctgccagga gacagcgtga ctctgacctg cccgggggta gagccggaag acaatgccac
     1261 tgttcactgg gtgctcagga agccggctgc aggctcccac cccagcagat gggctggcat
     1321 gggaaggagg ctgctgctga ggtcggtgca gctccacgac tctggaaact attcatgcta
     1381 ccgggccggc cgcccagctg ggactgtgca cttgctggtg gatgttcccc ccgaggagcc
     1441 ccagctctcc tgcttccgga agagccccct cagcaatgtt gtttgtgagt ggggtcctcg
     1501 gagcacccca tccctgacga caaaggctgt gctcttggtg aggaagtttc agaacagtcc
     1561 ggccgaagac ttccaggagc cgtgccagta ttcccaggag tcccagaagt tctcctgcca
     1621 gttagcagtc ccggagggag acagctcttt ctacatagtg tccatgtgcg tcgccagtag
     1681 tgtcgggagc aagttcagca aaactcaaac ctttcagggt tgtggaatct tgcagcctga
     1741 tccgcctgcc aacatcacag tcactgccgt ggccagaaac ccccgctggc tcagtgtcac
     1801 ctggcaagac ccccactcct ggaactcatc tttctacaga ctacggtttg agctcagata
     1861 tcgggctgaa cggtcaaaga cattcacaac atggatggtc aaggacctcc agcatcactg
     1921 tgtcatccac gacgcctgga gcggcctgag gcacgtggtg cagcttcgtg cccaggagga
     1981 gttcgggcaa ggcgagtgga gcgagtggag cccggaggcc atgggcacgc cttggacaga
     2041 atccaggagt cctccagctg agaacgaggt gtccaccccc atgcaggcac ttactactaa
     2101 taaagacgat gataatattc tcttcagaga ttctgcaaat gcgacaagcc tcccaggagg
     2161 cagcgcggcc gcaggcgact acaaagacca tgacggtgat tataaagatc atgacattga
     2221 ttacaaggat gacgatgaca agtaataaga attctcgacc tcgagacaaa tggcagtatt
     2281 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2341 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2401 aaattttcgg gtttattaca gggacagcag agatccactt tggccgcggc tcgagggggt
     2461 tggggttgcg ccttttccaa ggcagccctg ggtttgcgca gggacgcggc tgctctgggc
     2521 gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc acgtccgttc
     2581 gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc cccggcgacg cttcctgctc
     2641 cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga caaacggaag
     2701 ccgcacgtct cactagtacc ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc
     2761 gaccgcgatg ggctgtggcc aatagcggct gctcagcagg gcgcgccgag agcagcggcc
     2821 gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg
     2881 cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg
     2941 accgaatcac cgacctctct ccccaggggg atccaccgga gcttaccatg aaaaagcctg
     3001 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     3061 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     3121 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     3181 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
     3241 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     3301 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     3361 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     3421 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     3481 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     3541 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     3601 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     3661 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     3721 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     3781 atatgctccg cattggtctt gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     3841 cgcacgaccc catgcatcgg tacctttaag accaatgact tacaaggcag ctgtagatct
     3901 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     3961 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     4021 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     4081 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     4141 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     4201 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     4261 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4321 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     4381 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4441 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4501 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     4561 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     4621 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     4681 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     4741 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4801 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4861 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4921 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4981 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5041 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5101 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5161 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     5221 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5281 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5341 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5401 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5461 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5521 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5581 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5641 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5701 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5761 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5821 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5881 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5941 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6001 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6061 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6121 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6181 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6241 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6301 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6361 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6421 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6481 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6541 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6601 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6661 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6721 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6781 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6841 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6901 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6961 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7021 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7081 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     7141 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     7201 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     7261 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     7321 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     7381 aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct
     7441 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
     7501 tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt
     7561 ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc
     7621 ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7681 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     7741 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     7801 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     7861 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     7921 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     7981 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     8041 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     8101 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     8161 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     8221 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     8281 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagca gcagctgatc
     8341 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     8401 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     8461 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     8521 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     8581 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     8641 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     8701 aaggatcaac agctcct
//
