LOCUS       sequence-388593-        6828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SLC7A11 with an N-terminal FLAG tag and C-terminal 
            TEV-GFP-His8 tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  AUTHORS   Parker JL, Deme JC, Kolokouris D, Kuteyi G, Biggin PC, Lea SM, 
            Newstead S
  TITLE     Molecular basis for redox control by the human cystine/glutamate 
            antiporter system xc().
  JOURNAL   Nat Commun. 2021 Dec 8;12(1):7147. doi: 10.1038/s41467-021-27414-1.
  PUBMED    34880232
REFERENCE   2  (bases 1 to 6828)
  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: "Nat 
            Commun."; date: "2021-12-8"; volume: "12(1):7147. doi"; pages: " 
            10.1038/s41467-021-27414-1"
COMMENT     Sequence Label: pLexM-SLC7A11_GFP
FEATURES             Location/Qualifiers
     source          1..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1774..1783
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1783..1806
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3316..3336
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3343..4053
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4054..4077
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(4183..4202)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4248..4303
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4302..4321)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4664..4680)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4664..4680)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4677..4699)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4688..4704
                     /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)."
     promoter        complement(join(4712..4718,4719..4736,4737..4742))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4757..4778
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4907..4924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5078..5666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5158..5177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5837..6628,6629..6697))
                     /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     6460..6479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6698..6802)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccrggsg gggsggggsg gggsgrgggg
      541 sggggsgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggkgggggt gccgggcggg
     1261 gcggggccgc ckcgggccgg ggrgggctcg ggggrggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgcggccg
     1741 tctcaggccg aattcaacgt tggtaccgag ctcgccacca tggactacaa agacgatgac
     1801 gacaaggtca gaaagcctgt tgtgtccacc atctccaaag gaggttacct gcagggaaat
     1861 gttaacggga ggctgccttc cctgggcaac aaggagccac ctgggcagga gaaagtgcag
     1921 ctgaagagga aagtcacttt actgagggga gtctccatta tcattggcac catcattgga
     1981 gcaggaatct tcatctctcc taagggcgtg ctccagaaca cgggcagcgt gggcatgtct
     2041 ctgaccatct ggacggtgtg tggggtcctg tcactatttg gagctttgtc ttatgctgaa
     2101 ttgggaacaa ctataaagaa atctggaggt cattacacat atattttgga agtctttggt
     2161 ccattaccag cttttgtacg agtctgggtg gaactcctca taatacgccc tgcagctact
     2221 gctgtgatat ccctggcatt tggacgctac attctggaac cattttttat tcaatgtgaa
     2281 atccctgaac ttgcgatcaa gctcattaca gctgtgggca taactgtagt gatggtccta
     2341 aatagcatga gtgtcagctg gagcgcccgg atccagattt tcttaacctt ttgcaagctc
     2401 acagcaattc tgataattat agtccctgga gttatgcagc taattaaagg tcaaacgcag
     2461 aactttaaag acgccttttc aggaagagat tcaagtatta cgcggttgcc actggctttt
     2521 tattatggaa tgtatgcata tgctggctgg ttttacctca actttgttac tgaagaagta
     2581 gaaaaccctg aaaaaaccat tccccttgca atatgtatat ccatggccat tgtcaccatt
     2641 ggctatgtgc tgacaaatgt ggcctacttt acgaccatta atgctgagga gctgctgctt
     2701 tcaaatgcag tggcagtgac cttttctgag cggctactgg gaaatttctc attagcagtt
     2761 ccgatctttg ttgccctctc ctgctttggc tccatgaacg gtggtgtgtt tgctgtctcc
     2821 aggttattct atgttgcgtc tcgagagggt caccttccag aaatcctctc catgattcat
     2881 gtccgcaagc acactcctct accagctgtt attgttttgc accctttgac aatgataatg
     2941 ctcttctctg gagacctcga cagtcttttg aatttcctca gttttgccag gtggcttttt
     3001 attgggctgg cagttgctgg gctgatttat cttcgataca aatgcccaga tatgcatcgt
     3061 cctttcaagg tgccactgtt catcccagct ttgttttcct tcacatgcct cttcatggtt
     3121 gccctttccc tctattcgga cccatttagt acagggattg gcttcgtcat cactctgact
     3181 ggagtccctg cgtattatct ctttattata tgggacaaga aacccaggtg gtttagaata
     3241 atgtcagaga aaataaccag aacattacaa ataatactgg aagttgtacc agaagaagat
     3301 aagttagtcg agggggaaaa tttatatttt caaggtcaat tttctaaagg tgaagaatta
     3361 ttcactggcg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt
     3421 tctgtctccg gtgaaggtga aggtgacgct acttacggta aattgacctt aaaatttatt
     3481 tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt cggttatggt
     3541 gttcaatgtt ttgcgagata cccagatcat atgaaacaac atgacttttt caagtctgcc
     3601 atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg taactacaag
     3661 accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt
     3721 attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct
     3781 cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt
     3841 agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca
     3901 attggtgatg gtccagtctt gttaccagac aaccattact tatccactca atctgcctta
     3961 tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct
     4021 ggtattaccc atggtatgga tgaattgtac aaacaccacc accatcacca tcatcattaa
     4081 gtcgagacta gtatcgcgat aattcactcc tcaggtgcag gctgcctatc agaaggtggt
     4141 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     4201 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     4261 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     4321 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     4381 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     4441 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     4501 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     4561 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     4621 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     4681 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4741 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4801 ctttccagtc gggaaacctg tcgtgccagc ggatcgatcc gctgcattaa tgaatcggcc
     4861 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4921 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4981 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5041 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5101 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5161 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5221 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5281 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5341 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5401 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5461 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5521 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5581 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5701 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5761 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5821 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5881 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5941 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6001 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6061 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6121 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6181 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6241 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6301 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6361 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6421 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6481 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6541 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6601 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6661 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6721 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6781 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
