LOCUS       sequence-200295-        7974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human GFP-ATIC fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7974)
  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 7974)
  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: pGFP-ATIC
FEATURES             Location/Qualifiers
     source          1..7974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        131..510
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        511..722
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     673..693
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     719..743
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          883..1015
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     998..1017
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1060..1078
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1111..1824
                     /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(1141..1169)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1764..1784
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    3671..3792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3682..3701)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3736..3755
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3978..4433
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4065..4084)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4275..4296
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4550..4907
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4550..4741
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4573..4593)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4758..4893
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4958..5752
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5012..5031)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5622..5641
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5816..5864
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5905..5924)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6024..6046
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6084..6102)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6170..6274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6275..6343,6344..7135)
                     /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(6493..6512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7306..7894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7795..7814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccattag ccatattatt cattggttat atagcataaa tcaatattgg ctattggcca
       61 ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga
      121 ccgccatgtt ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      181 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      241 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      301 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      361 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa
      421 tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac
      481 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg
      541 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      601 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc
      661 ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      721 ctcgtttagt gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa
      781 ttgctaacgc agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc
      841 ttaaggtagc cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca
      901 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt
      961 ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc
     1021 cactcccagt tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct
     1081 agcctcgagg caagtactgg ttcggaattc atgagtaaag gagaagaact tttcactgga
     1141 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
     1201 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
     1261 ggaaaactac ctgttccatg gccaacactt gtcactactt tcacttatgg tgttcaatgc
     1321 ttttcaagat acccagatca catgaaacag catgactttt tcaagagtgc catgcccgaa
     1381 ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct
     1441 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
     1501 aaagaagatg gaaacattct tggacacaaa ttggaatata actataactc acacaatgta
     1561 tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac
     1621 attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat
     1681 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
     1741 cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca
     1801 catggcatgg atgaactata caaagtaata aggatccggt cagtgatggc tcccggccag
     1861 ctcgccttat ttagtgtctc tgacaaaacc ggccttgtgg aatttgcaag aaacctgacc
     1921 gctcttggtt tgaatctggt cgcttccgga gggactgcaa aagctctcag ggatgctggt
     1981 ctggcagtca gagatgtctc tgagttgacg ggatttcctg aaatgttggg gggacgtgtg
     2041 aaaactttgc atcctgcagt ccatgctgga atcctagctc gtaatattcc agaagataat
     2101 gctgacatgg ccagacttga tttcaatctt ataagagttg ttgcctgcaa tctctatccc
     2161 tttgtaaaga cagtggcttc tccaggtgta actgttgagg aggctgtgga gcaaattgac
     2221 attggtggag taaccttact gagagctgca gccaaaaacc acgctcgagt gacagtggtg
     2281 tgtgaaccag aggactatgt ggtggtgtcc acggagatgc agagctccga gagtaaggac
     2341 acctccttgg agactagacg ccagttagcc ttgaaggcat tcactcatac ggcacaatat
     2401 gatgaagcaa tttcagatta tttcaggaaa cagtacagca aaggcgtatc tcagatgccc
     2461 ttgagatatg gaatgaaccc acatcagacc cctgcccagc tgtacacact gcagcccaag
     2521 cttcccatca cagttctaaa tggagcccct ggatttataa acttgtgcga tgctttgaac
     2581 gcctggcagc tggtgaagga actcaaggag gctttaggta ttccagccgc tgcctctttc
     2641 aaacatgtca gcccagcagg tgctgctgtt ggaattccac tcagtgaaga tgaggccaaa
     2701 gtctgcatgg tttatgatct ctataaaacc ctcacaccca tctcagcggc gtatgcaaga
     2761 gcaagagggg ctgataggat gtcttcattt ggtgattttg ttgcattgtc cgatgtttgt
     2821 gatgtaccaa ctgcaaaaat tatttccaga gaagtatctg atggtataat tgccccagga
     2881 tatgaagaag aagccttgac aatactttcc aaaaagaaaa atggaaacta ttgtgtcctt
     2941 cagatggacc aatcttacaa accagatgaa aatgaagttc gaactctctt tggtcttcat
     3001 ttaagccaga agagaaataa tggtgtcgtc gacaagtcat tatttagcaa tgttgttacc
     3061 aaaaataaag atttgccaga gtctgccctc cgagacctca tcgtagccac cattgctgtc
     3121 aagtacactc agtctaactc tgtgtgctac gccaagaacg ggcaggttat cggcattgga
     3181 gcaggacagc agtctcgtat acactgcact cgccttgcag gagataaggc aaactattgg
     3241 tggcttagac accatccaca agtgctttcg atgaagttta aaacaggagt gaagagagca
     3301 gaaatctcca atgccatcga tcaatatgtg actggaacca ttggcgagga tgaagatttg
     3361 ataaagtgga aggcactgtt tgaggaagtc cctgagttac tcactgaggc agagaagaag
     3421 gaatgggttg agaaactgac tgaagtttct atcagctctg atgccttctt ccctttccga
     3481 gataacgtag acagagctaa aaggagtggt gtggcgtaca ttgcggctcc ctccggttct
     3541 gctgctgaca aagttgtgat tgaggcctgc gacgaactgg gaatcatcct cgctcatacg
     3601 aaccttcggc tcttccacca ctaagcggcc gcttcccttt agtgagggtt aatgcttcga
     3661 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3721 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3781 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     3841 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatccg ataaggatcg
     3901 atccgggctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3961 gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4021 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4081 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4141 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4201 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4261 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4321 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4381 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg
     4441 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag
     4501 caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa
     4561 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4621 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4681 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4741 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4801 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4861 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgattctt
     4921 ctgacacaac agtctcgaac ttaaggctag agccaccatg attgaacaag atggattgca
     4981 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5041 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5101 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5161 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5221 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5281 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5341 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5401 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5461 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5521 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5581 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5641 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5701 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5761 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     5821 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     5881 catccgcgta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     5941 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6001 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6061 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     6121 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6181 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6241 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6301 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6361 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6421 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6481 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6541 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6601 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6661 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6721 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6781 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6841 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6901 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6961 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7021 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7081 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7141 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7201 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7321 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7381 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7441 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7501 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7561 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7621 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7681 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7741 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7801 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7861 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7921 gttcctggcc ttttgctggc cttttgctca catggctcga cagatcttca atat
//
