LOCUS       sequence-388568-        7026 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses SLC3A2 as an N-terminal His tagged construct in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7026)
  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 7026)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-SLC3A2
FEATURES             Location/Qualifiers
     source          1..7026
                     /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"
     CDS             1777..1800
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3514..3534
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3541..4251
                     /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             4252..4275
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(4381..4400)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4446..4501
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4500..4519)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4862..4878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4862..4878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4875..4897)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4886..4902
                     /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(4910..4916,4917..4934,4935..4940))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4955..4976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5105..5122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5276..5864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5356..5375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6035..6826,6827..6895))
                     /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     6658..6677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6896..7000)
                     /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 tggtaccgcc accatgcacc atcaccatca ccatcaccat
     1801 gagctacagc ctcctgaagc ctcgatcgcc gtcgtgtcga ttccgcgcca gttgcctggc
     1861 tcacattcgg aggctggtgt ccagggtctc agcgcggggg acgactcagg caccatgagc
     1921 caggacaccg aggtggatat gaaggaggtg gagctgaatg agttagagcc cgagaagcag
     1981 ccgatgaacg cggcgtctgg ggcggccatg tccctggcgg gagccgagaa gaatggtctg
     2041 gtgaagatca aggtggcgga agacgaggcg gaggcggcag ccgcggctaa gttcacgggc
     2101 ctgtccaagg aggagctgct gaaggtggca ggcagccccg gctgggtacg cacccgctgg
     2161 gcactgctgc tgctcttctg gctcggctgg ctcggcatgc ttgctggtgc cgtggtcata
     2221 atcgtgcgag cgccgcgttg tcgcgagcta ccggcgcaga agtggtggca cacgggcgcc
     2281 ctctaccgca tcggcgacct tcaggccttc cagggccacg gcgcgggcaa cctggcgggt
     2341 ctgaaggggc gtctcgatta cctgagctct ctgaaggtga agggccttgt gctgggtcca
     2401 attcacaaga accagaagga tgatgtcgct cagactgact tgctgcagat cgaccccaat
     2461 tttggctcca aggaagattt tgacagtctc ttgcaatcgg ctaaaaaaaa gagcatccgt
     2521 gtcattctgg accttactcc caactaccgg ggtgagaact cgtggttctc cactcaggtt
     2581 gacactgtgg ccaccaaggt gaaggatgct ctggagtttt ggctgcaagc tggcgtggat
     2641 gggttccagg ttcgggacat agagaatctg aaggatgcat cctcattctt ggctgagtgg
     2701 caaaatatca ccaagggctt cagtgaagac aggctcttga ttgcggggac taactcctcc
     2761 gaccttcagc agatcctgag cctactcgaa tccaacaaag acttgctgtt gactagctca
     2821 tacctgtctg attctggttc tactggggag catacaaaat ccctagtcac acagtatttg
     2881 aatgccactg gcaatcgctg gtgcagctgg agtttgtctc aggcaaggct cctgacttcc
     2941 ttcttgccgg ctcaacttct ccgactctac cagctgatgc tcttcaccct gccagggacc
     3001 cctgttttca gctacgggga tgagattggc ctggatgcag ctgcccttcc tggacagcct
     3061 atggaggctc cagtcatgct gtgggatgag tccagcttcc ctgacatccc aggggctgta
     3121 agtgccaaca tgactgtgaa gggccagagt gaagaccctg gctccctcct ttccttgttc
     3181 cggcggctga gtgaccagcg gagtaaggag cgctccctac tgcatgggga cttccacgcg
     3241 ttctccgctg ggcctggact cttctcctat atccgccact gggaccagaa tgagcgtttt
     3301 ctggtagtgc ttaactttgg ggatgtgggc ctctcggctg gactgcaggc ctccgacctg
     3361 cctgccagcg ccagcctgcc agccaaggct gacctcctgc tcagcaccca gtcaggccgt
     3421 gaggagggct cccctcttga gctggaacgc ctgaaactgg agcctcacga agggctgctg
     3481 ctccgcttcc cctacgcggc ctgactcgag ggggaaaatt tatattttca aggtcaattt
     3541 tctaaaggtg aagaattatt cactggcgtt gtcccaattt tggttgaatt agatggtgat
     3601 gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgacgctac ttacggtaaa
     3661 ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc aaccttagtc
     3721 actactttcg gttatggtgt tcaatgtttt gcgagatacc cagatcatat gaaacaacat
     3781 gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa
     3841 gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat
     3901 agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg
     3961 gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa gaatggtatc
     4021 aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt agctgaccat
     4081 tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta
     4141 tccactcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta
     4201 gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa acaccaccac
     4261 catcaccatc atcattaagt cgagactagt atcgcgataa ttcactcctc aggtgcaggc
     4321 tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga
     4381 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     4441 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     4501 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     4561 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     4621 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     4681 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     4741 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     4801 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
     4861 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4921 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4981 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atcgatccgc
     5041 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5101 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5161 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5221 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5281 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5341 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5401 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5461 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5521 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5581 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5641 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5701 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5761 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5821 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5881 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5941 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6001 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6061 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6121 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6181 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6241 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6301 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6361 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6421 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6481 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6541 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6601 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6661 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6721 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6781 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6841 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6901 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6961 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7021 cacctg
//
