LOCUS       sequence-200317-        9236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human GART-GFP fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9236)
  AUTHORS   An S, Kumar R, Sheets ED, Benkovic SJ
  TITLE     Reversible compartmentalization of de novo purine biosynthetic 
            complexes in living cells.
  JOURNAL   Science. 2008 Apr 4;320(5872):103-6. doi: 10.1126/science.1152241.
  PUBMED    18388293
REFERENCE   2  (bases 1 to 9236)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2008-04-4- 4"; volume: "320"; issue: 
            "5872"; pages: "103-6"
COMMENT     Sequence Label: pGART-GFP
FEATURES             Location/Qualifiers
     source          1..9236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..711
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     708..732
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          872..1004
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     987..1006
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1049..1067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4130..4843
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4160..4188)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4783..4803
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    4939..5060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4950..4969)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5004..5023
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5246..5701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5333..5352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5543..5564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5818..6175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5818..6009
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5841..5861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6026..6161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6280..6299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6873..6892
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7067..7115
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(7156..7175)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7275..7297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7335..7353)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7421..7525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7526..7594,7595..8386)
                     /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(7744..7763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8557..9145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9046..9065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catattattc attggttata tagcataaat caatattggc tattggccat tgcatacgtt
       61 gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg
      121 gcattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tgcgatcgcc cgccccgttg
      661 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      721 aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat tgctaacgca
      781 gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gtgactctct taaggtagcc
      841 ttgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      901 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      961 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt
     1021 caattacagc tcttaaggct agagtactta atacgactca ctataggcta gcatggctgc
     1081 tcgtgtactt ataattggca gtggaggaag ggaacatacg ctggcctgga aacttgcaca
     1141 gtctcatcat gtcaaacaag tgttggttgc cccaggaaac gcaggcactg cctgctctga
     1201 aaagatttca aataccgcca tctcaatcag tgaccacact gcccttgctc aattctgcaa
     1261 agagaagaaa attgaatttg tagttgttgg accagaagca cctctggctg ctgggattgt
     1321 tgggaacctg aggtctgcag gagtgcaatg ctttggccca acagcagaag cggctcagtt
     1381 agagtccagc aaaaggtttg ccaaagagtt tatggacaga catggaatcc caaccgcaca
     1441 atggaaggct ttcaccaaac ctgaagaagc ctgcagcttc attttgagtg cagacttccc
     1501 tgctttggtt gtgaaggcca gtggtcttgc agctggaaaa ggggtgattg ttgcaaagag
     1561 caaagaagag gcctgcaaag ctgtacaaga gatcatgcag gagaaagcct ttggggcagc
     1621 tggagaaaca attgtcattg aagaacttct tgacggagaa gaggtgtcgt gtctgtgttt
     1681 cactgatggc aagactgtgg cccccatgcc cccagcacag gaccataagc gattactgga
     1741 gggagatggt ggccctaaca cagggggaat gggagcctat tgtccagccc ctcaggtttc
     1801 taatgatcta ttactaaaaa ttaaagatac tgttcttcag aggacagtgg atggcatgca
     1861 gcaagagggt actccatata caggtattct ctatgctgga ataatgctga ccaagaatgg
     1921 cccaaaagtt ctagagttta attgccgttt tggtgatcca gagtgccaag taatcctccc
     1981 acttcttaaa agtgaccttt atgaagtgat tcagtccacc ttagatggac tgctctgcac
     2041 atctctgcct gtttggctag aaaaccacac cgccctaact gttgtcatgg caagtaaagg
     2101 ttatcctgga gactacacca agggtgtaga gataacaggg tttcctgagg ctcaagctct
     2161 aggactggag gtgttccatg caggcactgc cctcaaaaat ggcaaagtag taactcatgg
     2221 gggtagagtt cttgcagtca cagccatccg ggaaaatctc atatcagccc ttgaggaagc
     2281 caagaaagga ctagctgcta taaagtttga gggagcaatt tataggaaag acgtcggctt
     2341 tcgtgccata gctttcctcc agcagcccag gagtttgact tacaaggaat ctggagtaga
     2401 tatcgcagct ggaaatatgc tggtcaagaa aattcagcct ttagcaaaag ccacttccag
     2461 atcaggctgt aaagttgatc ttggaggttt tgctggtctt tttgatttaa aagcagctgg
     2521 tttcaaagat ccccttctgg cctctggaac agatggcgtt ggaactaaac taaagattgc
     2581 ccagctatgc aataaacatg ataccattgg tcaagatttg gtagcaatgt gtgttaatga
     2641 tattctggca caaggagcag agcccctctt cttccttgat tacttttcct gtggaaaact
     2701 tgacctcagt gtaactgaag ctgttgttgc tggaattgct aaagcttgtg gaaaagctgg
     2761 atgtgctctc cttggaggtg aaacagcaga aatgcctgac atgtatcccc ctggagagta
     2821 tgacctagct gggtttgccg ttggtgccat ggagcgagat cagaaactcc ctcacctgga
     2881 aagaatcact gagggtgatg ttgttgttgg aatagcttca tctggtcttc atagcaatgg
     2941 atttagcctt gtgaggaaaa tcgtggcaaa atcttccctc cagtactcct ctccagcacc
     3001 tgatggttgt ggtgaccaga ctttagggga cttacttctc acgcctacca gaatctacag
     3061 ccattcactg ttacctgtcc tacgttcagg acatgtcaaa gcctttgccc atattactgg
     3121 tggaggatta ctagagaaca tccccagagt cctccctgag aaacttgggg tagatttaga
     3181 tgcccagacc tggaggatcc ccagggtctt ctcatggttg cagcaggaag gacacctctc
     3241 tgaggaagag atggccagaa catttaactg tggggttggc gctgtccttg tggtatcaaa
     3301 ggagcagaca gagcagattc tgagggatat ccagcagcac aaggaagaag cctgggtgat
     3361 tggcagtgtg gttgcacgag ctgaaggttc cccacgtgtg aaagtcaaga atctgattga
     3421 aagcatgcaa ataaatgggt cagtgttgaa gaatggctcc ctgacaaatc atttctcttt
     3481 tgaaaaaaaa aaggccagag tggctgtctt aatatctgga acaggatcga acctgcaagc
     3541 acttatagac agtactcggg aaccaaatag ctctgcacaa attgatattg ttatctccaa
     3601 caaagccgca gtagctgggt tagataaagc ggaaagagct ggtattccca ctagagtaat
     3661 taatcataaa ctgtataaaa atcgtgtaga atttgacagt gcaattgacc tagtccttga
     3721 agagttctcc atagacatag tctgtcttgc aggattcatg agaattcttt ctggcccctt
     3781 tgtccaaaag tggaatggaa aaatgctcaa tatccaccca tccttgctcc cttcttttaa
     3841 gggttcaaat gcccatgagc aagccctgga aaccggagtc acagttactg ggtgcactgt
     3901 acactttgta gctgaagatg tggatgctgg acagattatt ttgcaagaag ctgttcccgt
     3961 gaagaggggt gatactgtcg caactctttc tgaaagagta aaattagcag aacataaaat
     4021 atttcctgca gcccttcagc tggtggccag tggaactgta cagcttggag aaaatggcaa
     4081 gatctgttgg gttaaagagg aactcgaggc aagtactggt tcggaattca tgagtaaagg
     4141 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
     4201 gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct
     4261 taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt
     4321 cacttatggt gttcaatgct tttcaagata cccagatcac atgaaacagc atgacttttt
     4381 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
     4441 gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga
     4501 gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat tggaatataa
     4561 ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa
     4621 cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca
     4681 aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca
     4741 atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt
     4801 aacagctgct gggattacac atggcatgga tgaactatac aaataataag gatcccagga
     4861 gctcggcaag cttgcggccg cactcgaccc gggcggccgc ttccctttag tgagggttaa
     4921 tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     4981 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     5041 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     5101 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat
     5161 aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     5221 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5281 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5341 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5401 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5461 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5521 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5581 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5641 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     5701 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     5761 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     5821 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5881 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5941 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6001 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6061 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6121 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     6181 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     6241 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6301 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6361 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     6421 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6481 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6541 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6601 cttgatccgg ctacctgccc attcgaccac caagcgagca cgtactcgga tggaagccgg
     6661 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6721 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6781 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6841 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6901 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6961 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     7021 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
     7081 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgcgt
     7141 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     7201 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     7261 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     7321 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     7381 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     7441 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     7501 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     7561 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     7621 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     7681 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     7741 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     7801 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     7861 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     7921 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     7981 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8041 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     8101 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     8161 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8221 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     8281 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8341 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     8401 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8461 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8521 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     8581 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     8641 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     8701 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     8761 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     8821 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     8881 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     8941 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     9001 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     9061 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     9121 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     9181 cttttgctgg ccttttgctc acatggctcg acagatcttc aatattggcc attagc
//
