LOCUS       sequence-318421-        8280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rescues the expression of GPX8 in GPX8 plenti-CRISPR KO cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8280)
  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 8280)
  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: GPX8 rescue PLAK1
FEATURES             Location/Qualifiers
     source          1..8280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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     1606..1626
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        1915..2118
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2069..2089
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2819..2884
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2932..3049
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3098..3608
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3527..3545
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     LTR             4572..4805
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4877..4998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4914..4933)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4968..4987
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5038..5173
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5193..5212)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5194..5212)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5222..5239)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5222..5238)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5231..5253)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5380..5835
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5467..5486)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5677..5698
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5861..5965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5966..6034,6035..6826)
                     /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(6184..6203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6997..7585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7486..7505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7739..7756
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7873..7894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7909..7914,7915..7932,7933..7939)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7947..7963
                     /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     7952..7974
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7971..7987
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7971..7987
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8006..8026
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8008..8026
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8054..8280
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cagcagctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcgat cacgagacta gcctcgagca gcagccccct tcaccgaggg cctatttccc
     1621 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     1681 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     1741 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgcc aagtacgccc
     1801 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1861 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1921 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1981 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     2041 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     2101 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtttaat
     2161 taagtccaac atggagcctc ttgcagctta cccgctaaaa tgttccgggc cacgcgctaa
     2221 ggtatttgca gttttgctgt ctatagttct atgcacagta acgctatttc ttctacaact
     2281 aaaattcctc aaacctaaaa tcaacagctt ttatgccttt gaagtgaagg atgcaaaagg
     2341 aagaactgtt tctctggaaa agtataaagg caaagtttca ctagttgtaa acgtggccag
     2401 tgactgccaa ctcacagaca gaaattactt agggctgaag gaactgcaca aagagtttgg
     2461 accatcccac ttcagcgtgt tggcttttcc ctgcaatcag tttggagaat cggagccccg
     2521 cccaagcaag gaagtagaat cttttgcaag aaaaaactac ggagtaactt tccccatctt
     2581 ccacaagatt aagattctag gatctgaagg agaacctgca tttagatttc ttgttgattc
     2641 ttcaaagaag gaaccaaggt ggaatttttg gaagtatctt gtcaaccctg agggtcaagt
     2701 tgtgaagttc tggaagccag aggagcccat tgaagtcatc aggcctgaca tagcagctct
     2761 ggttagacaa gtgatcataa aaaagaaaga ggatctagga ggcagcgcgg ccgcaggcga
     2821 ctacaaagac catgacggtg attataaaga tcatgacatt gattacaagg atgacgatga
     2881 caagtaataa gaattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag
     2941 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     3001 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     3061 cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg cgccttttcc
     3121 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
     3181 gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct
     3241 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
     3301 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
     3361 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
     3421 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
     3481 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
     3541 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
     3601 ctccccaggg ggatccaccg gagcttacca tgaaaaagcc tgaactcacc gcgacgtctg
     3661 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     3721 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     3781 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
     3841 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
     3901 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     3961 ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
     4021 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     4081 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     4141 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     4201 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     4261 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     4321 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     4381 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     4441 ttgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc
     4501 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     4561 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     4621 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     4681 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     4741 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     4801 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     4861 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4921 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4981 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     5041 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5101 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5161 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     5221 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     5281 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     5341 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     5401 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5461 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5521 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5581 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5641 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5701 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5761 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5821 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5881 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5941 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6001 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6061 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6121 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6181 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6241 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6301 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6361 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6421 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6481 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6541 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6601 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6661 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6721 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6781 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6841 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6901 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6961 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7021 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7081 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7141 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7201 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7261 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7321 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7381 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7441 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7501 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7561 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7621 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7681 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7741 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     7801 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     7861 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     7921 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     7981 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     8041 gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg
     8101 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta
     8161 aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga
     8221 accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac
//
