LOCUS       sequence-193176-        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        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     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     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             1118..1831
                     /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(1148..1176)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1771..1791
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    3678..3799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3689..3708)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3743..3762
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3985..4440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4072..4091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4282..4303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4557..4914
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4557..4748
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4580..4600)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4765..4900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4965..5759
                     /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(5019..5038)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5629..5648
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5823..5871
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5912..5931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6031..6053
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6091..6109)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6177..6281
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6282..6350,6351..7142)
                     /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(6500..6519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7313..7901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7802..7821
                     /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 ctcgaggcaa gtactggttc ggaattcatg agtaaaggag aagaactttt
     1141 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
     1201 tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg
     1261 cactactgga aaactacctg ttccatggcc aacacttgtc actactttca cttatggtgt
     1321 tcaatgcttt tcaagatacc cagatcacat gaaacagcat gactttttca agagtgccat
     1381 gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac
     1441 acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat
     1501 tgattttaaa gaagatggaa acattcttgg acacaaattg gaatataact ataactcaca
     1561 caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact tcaaaattag
     1621 acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     1681 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     1741 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     1801 gattacacat ggcatggatg aactatacaa agtaataagg atccggtcag tgatggctcc
     1861 cggccagctc gccttattta gtgtctctga caaaaccggc cttgtggaat ttgcaagaaa
     1921 cctgaccgct cttggtttga atctggtcgc ttccggaggg actgcaaaag ctctcaggga
     1981 tgctggtctg gcagtcagag atgtctctga gttgacggga tttcctgaaa tgttgggggg
     2041 acgtgtgaaa actttgcatc ctgcagtcca tgctggaatc ctagctcgta atattccaga
     2101 agataatgct gacatggcca gacttgattt caatcttata agagttgttg cctgcaatct
     2161 ctatcccttt gtaaagacag tggcttctcc aggtgtaact gttgaggagg ctgtggagca
     2221 aattgacatt ggtggagtaa ccttactgag agctgcagcc aaaaaccacg ctcgagtgac
     2281 agtggtgtgt gaaccagagg actatgtggt ggtgtccacg gagatgcaga gctccgagag
     2341 taaggacacc tccttggaga ctagacgcca gttagccttg aaggcattca ctcatacggc
     2401 acaatatgat gaagcaattt cagattattt caggaaacag tacagcaaag gcgtatctca
     2461 gatgcccttg agatatggaa tgaacccaca tcagacccct gcccagctgt acacactgca
     2521 gcccaagctt cccatcacag ttctaaatgg agcccctgga tttataaact tgtgcgatgc
     2581 tttgaacgcc tggcagctgg tgaaggaact caaggaggct ttaggtattc cagccgctgc
     2641 ctctttcaaa catgtcagcc cagcaggtgc tgctgttgga attccactca gtgaagatga
     2701 ggccaaagtc tgcatggttt atgatctcta taaaaccctc acacccatct cagcggcgta
     2761 tgcaagagca agaggggctg ataggatgtc ttcatttggt gattttgttg cattgtccga
     2821 tgtttgtgat gtaccaactg caaaaattat ttccagagaa gtatctgatg gtataattgc
     2881 cccaggatat gaagaagaag ccttgacaat actttccaaa aagaaaaatg gaaactattg
     2941 tgtccttcag atggaccaat cttacaaacc agatgaaaat gaagttcgaa ctctctttgg
     3001 tcttcattta agccagaaga gaaataatgg tgtcgtcgac aagtcattat ttagcaatgt
     3061 tgttaccaaa aataaagatt tgccagagtc tgccctccga gacctcatcg tagccaccat
     3121 tgctgtcaag tacactcagt ctaactctgt gtgctacgcc aagaacgggc aggttatcgg
     3181 cattggagca ggacagcagt ctcgtataca ctgcactcgc cttgcaggag ataaggcaaa
     3241 ctattggtgg cttagacacc atccacaagt gctttcgatg aagtttaaaa caggagtgaa
     3301 gagagcagaa atctccaatg ccatcgatca atatgtgact ggaaccattg gcgaggatga
     3361 agatttgata aagtggaagg cactgtttga ggaagtccct gagttactca ctgaggcaga
     3421 gaagaaggaa tgggttgaga aactgactga agtttctatc agctctgatg ccttcttccc
     3481 tttccgagat aacgtagaca gagctaaaag gagtggtgtg gcgtacattg cggctccctc
     3541 cggttctgct gctgacaaag ttgtgattga ggcctgcgac gaactgggaa tcatcctcgc
     3601 tcatacgaac cttcggctct tccaccacta agcggccgct tccctttagt gagggttaat
     3661 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3721 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3781 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3841 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatccgata
     3901 aggatcgatc cgggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3961 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4021 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4081 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4141 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4201 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4261 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4321 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4381 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4441 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc
     4501 tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg taccttctga
     4561 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4621 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4681 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4741 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4801 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4861 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     4921 gattcttctg acacaacagt ctcgaactta aggctagagc caccatgatt gaacaagatg
     4981 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5041 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5101 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5161 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5221 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5281 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5341 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5401 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5461 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5521 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5581 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5641 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5701 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5761 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc
     5821 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     5881 cgataaggat ccgcgtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5941 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     6001 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     6061 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     6121 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     6181 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6241 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6301 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6361 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6421 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6481 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6541 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6601 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6661 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6721 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6781 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6841 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6901 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6961 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7021 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7081 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     7141 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     7201 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7261 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7321 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7381 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7441 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7501 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7561 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7621 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7681 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7741 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7801 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7861 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7921 ttttacggtt cctggccttt tgctggcctt ttgctcacat ggctcgacag atct
//
