LOCUS       sequence-256585-        8551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human GART protein with a C-terminal 2XStrep affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8551)
  AUTHORS   Liu C, Knudsen GM, Pedley AM, He J, Johnson JL, Yaron TM, Cantley 
            LC, Benkovic SJ
  TITLE     Mapping Post-Translational Modifications of de Novo Purine 
            Biosynthetic Enzymes: Implications for Pathway Regulation.
  JOURNAL   J Proteome Res. 2019 May 3;18(5):2078-2087. doi: 
            10.1021/acs.jproteome.8b00969. Epub 2019 Apr 18.
  PUBMED    30964683
REFERENCE   2  (bases 1 to 8551)
  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: 
            "10.1021/acs.jproteome.8b00969"; journalName: "J Proteome Res"; 
            date: "2019-05-3- 3"; volume: "18"; issue: "5"; pages: "2078-2087"
COMMENT     Sequence Label: pGART-2XStrep
FEATURES             Location/Qualifiers
     source          1..8551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4130..4135,4136..4159,4160..4195,4196..4219)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    4272..4393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4283..4302)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4337..4356
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4579..5034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4666..4685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4876..4897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5151..5508
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5151..5342
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5174..5194)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5359..5494
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5613..5632)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6206..6225
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6400..6448
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6489..6508)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6608..6630
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6668..6686)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6754..6858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6859..6927,6928..7719)
                     /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(7077..7096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7890..8478
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8379..8398
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc atggctgctc gtgtacttat aattggcagt ggaggaaggg aacatacgct
     1141 ggcctggaaa cttgcacagt ctcatcatgt caaacaagtg ttggttgccc caggaaacgc
     1201 aggcactgcc tgctctgaaa agatttcaaa taccgccatc tcaatcagtg accacactgc
     1261 ccttgctcaa ttctgcaaag agaagaaaat tgaatttgta gttgttggac cagaagcacc
     1321 tctggctgct gggattgttg ggaacctgag gtctgcagga gtgcaatgct ttggcccaac
     1381 agcagaagcg gctcagttag agtccagcaa aaggtttgcc aaagagttta tggacagaca
     1441 tggaatccca accgcacaat ggaaggcttt caccaaacct gaagaagcct gcagcttcat
     1501 tttgagtgca gacttccctg ctttggttgt gaaggccagt ggtcttgcag ctggaaaagg
     1561 ggtgattgtt gcaaagagca aagaagaggc ctgcaaagct gtacaagaga tcatgcagga
     1621 gaaagccttt ggggcagctg gagaaacaat tgtcattgaa gaacttcttg acggagaaga
     1681 ggtgtcgtgt ctgtgtttca ctgatggcaa gactgtggcc cccatgcccc cagcacagga
     1741 ccataagcga ttactggagg gagatggtgg ccctaacaca gggggaatgg gagcctattg
     1801 tccagcccct caggtttcta atgatctatt actaaaaatt aaagatactg ttcttcagag
     1861 gacagtggat ggcatgcagc aagagggtac tccatataca ggtattctct atgctggaat
     1921 aatgctgacc aagaatggcc caaaagttct agagtttaat tgccgttttg gtgatccaga
     1981 gtgccaagta atcctcccac ttcttaaaag tgacctttat gaagtgattc agtccacctt
     2041 agatggactg ctctgcacat ctctgcctgt ttggctagaa aaccacaccg ccctaactgt
     2101 tgtcatggca agtaaaggtt atcctggaga ctacaccaag ggtgtagaga taacagggtt
     2161 tcctgaggct caagctctag gactggaggt gttccatgca ggcactgccc tcaaaaatgg
     2221 caaagtagta actcatgggg gtagagttct tgcagtcaca gccatccggg aaaatctcat
     2281 atcagccctt gaggaagcca agaaaggact agctgctata aagtttgagg gagcaattta
     2341 taggaaagac gtcggctttc gtgccatagc tttcctccag cagcccagga gtttgactta
     2401 caaggaatct ggagtagata tcgcagctgg aaatatgctg gtcaagaaaa ttcagccttt
     2461 agcaaaagcc acttccagat caggctgtaa agttgatctt ggaggttttg ctggtctttt
     2521 tgatttaaaa gcagctggtt tcaaagatcc ccttctggcc tctggaacag atggcgttgg
     2581 aactaaacta aagattgccc agctatgcaa taaacatgat accattggtc aagatttggt
     2641 agcaatgtgt gttaatgata ttctggcaca aggagcagag cccctcttct tccttgatta
     2701 cttttcctgt ggaaaacttg acctcagtgt aactgaagct gttgttgctg gaattgctaa
     2761 agcttgtgga aaagctggat gtgctctcct tggaggtgaa acagcagaaa tgcctgacat
     2821 gtatccccct ggagagtatg acctagctgg gtttgccgtt ggtgccatgg agcgagatca
     2881 gaaactccct cacctggaaa gaatcactga gggtgatgtt gttgttggaa tagcttcatc
     2941 tggtcttcat agcaatggat ttagccttgt gaggaaaatc gtggcaaaat cttccctcca
     3001 gtactcctct ccagcacctg atggttgtgg tgaccagact ttaggggact tacttctcac
     3061 gcctaccaga atctacagcc attcactgtt acctgtccta cgttcaggac atgtcaaagc
     3121 ctttgcccat attactggtg gaggattact agagaacatc cccagagtcc tccctgagaa
     3181 acttggggta gatttagatg cccagacctg gaggatcccc agggtcttct catggttgca
     3241 gcaggaagga cacctctctg aggaagagat ggccagaaca tttaactgtg gggttggcgc
     3301 tgtccttgtg gtatcaaagg agcagacaga gcagattctg agggatatcc agcagcacaa
     3361 ggaagaagcc tgggtgattg gcagtgtggt tgcacgagct gaaggttccc cacgtgtgaa
     3421 agtcaagaat ctgattgaaa gcatgcaaat aaatgggtca gtgttgaaga atggctccct
     3481 gacaaatcat ttctcttttg aaaaaaaaaa ggccagagtg gctgtcttaa tatctggaac
     3541 aggatcgaac ctgcaagcac ttatagacag tactcgggaa ccaaatagct ctgcacaaat
     3601 tgatattgtt atctccaaca aagccgcagt agctgggtta gataaagcgg aaagagctgg
     3661 tattcccact agagtaatta atcataaact gtataaaaat cgtgtagaat ttgacagtgc
     3721 aattgaccta gtccttgaag agttctccat agacatagtc tgtcttgcag gattcatgag
     3781 aattctttct ggcccctttg tccaaaagtg gaatggaaaa atgctcaata tccacccatc
     3841 cttgctccct tcttttaagg gttcaaatgc ccatgagcaa gccctggaaa ccggagtcac
     3901 agttactggg tgcactgtac actttgtagc tgaagatgtg gatgctggac agattatttt
     3961 gcaagaagct gttcccgtga agaggggtga tactgtcgca actctttctg aaagagtaaa
     4021 attagcagaa cataaaatat ttcctgcagc ccttcagctg gtggccagtg gaactgtaca
     4081 gcttggagaa aatggcaaga tctgttgggt taaagaggaa ctcgagggga gcgcttggag
     4141 ccacccgcag ttcgaaaaag gtggaggttc tggcggtgga tcgggaggtt cagcgtggag
     4201 ccacccgcag ttcgagaaat aaagcgcggc cgcttccctt tagtgagggt taatgcttcg
     4261 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4321 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4381 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     4441 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcc gataaggatc
     4501 gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4561 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4621 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4681 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4741 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4801 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4861 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4921 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4981 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     5041 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     5101 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     5161 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5221 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5281 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5341 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5401 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5461 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct
     5521 tctgacacaa cagtctcgaa cttaaggcta gagccaccat gattgaacaa gatggattgc
     5581 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5641 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5701 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5761 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5821 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5881 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5941 cggctacctg cccattcgac caccaagcga gcacgtactc ggatggaagc cggtcttgtc
     6001 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6061 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6121 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6181 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6241 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6301 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6361 ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca
     6421 ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc
     6481 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     6541 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6601 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     6661 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6721 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6781 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6841 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6901 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6961 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7021 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7081 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     7141 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     7201 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7261 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7321 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7381 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     7441 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7501 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7561 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7621 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7681 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7741 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7801 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7861 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7921 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7981 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     8041 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     8101 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     8161 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     8221 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     8281 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     8341 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8401 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8461 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8521 tggccttttg ctcacatggc tcgacagatc t
//
