LOCUS       sequence-197930-        5977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human PAICS-EGFP fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  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 5977)
  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: pPAICS-EGFP
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        361..564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     515..535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1913..1922
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1919..1921,1922..1924,1925..2638)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLK
                     ERVTAAGITLGMDELYK"
     primer_bind     complement(1964..1985)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2225..2244)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2761..2882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2798..2817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2852..2871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2889..3344)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3026..3047)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3238..3257
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3371..3475
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3477..3834
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3477..3668
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3500..3520)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3685..3820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3869..4663
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3869..4663
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3923..3942)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4533..4552
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4851..4870)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4895..4942
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5271..5859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5760..5779
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
       61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      541 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcatggcgac
      601 agctgaggta ctgaacattg gtaaaaaatt atatgagggt aaaacaaaag aagtctacga
      661 attgttagac agtccaggaa aagtcctcct gcagtccaag gaccagatta cagcaggaaa
      721 tgcagctaga aaaaaccacc tggaaggaaa agctgcaatc tcaaataaaa tcaccagttg
      781 tatttttcag ttattacagg aagcaggtat taaaactgcc ttcaccagaa aatgtgggga
      841 gacagctttc attgcaccgc agtgtgaaat gattccaatt gaatgggttt gcagaagaat
      901 agcaactggt tcttttctca aaagaaatcc tggtgtcaag gaaggatata agttttaccc
      961 acctaaagtg gagttgtttt tcaaggatga tgccaataat gacccacagt ggtctgagga
     1021 acagctgatt gctgcaaaat tttgctttgc tggacttctt ataggccaga ctgaagtgga
     1081 tatcatgagt catgctacac aggctatatt tgaaatactg gagaaatcct ggttgcccca
     1141 gaattgtaca ctggttgata tgaagattga atttggtgtt gatgtaacca ccaaagaaat
     1201 tgttcttgct gatgttattg acaatgattc ctggagactc tggccatcag gagatcgaag
     1261 ccaacagaaa gacaaacagt cttatcggga cctcaaagaa gtaactcctg aagggctcca
     1321 aatggtaaag aaaaactttg agtgggttgc agagagagta gagttgcttt tgaaatcaga
     1381 aagtcagtgc agggttgtag tgttgatggg ctctacttct gatcttggtc actgtgaaaa
     1441 aatcaagaag gcctgtggaa attttggcat tccatgtgaa cttcgagtaa catctgcgca
     1501 taaaggacca gatgaaactc tgaggattaa agctgagtat gaaggggatg gcattcctac
     1561 tgtatttgtg gcagtggcag gcagaagtaa tggtttggga ccagtgatgt ctgggaacac
     1621 tgcatatcca gttatcagct gtcctcccct cacaccagac tggggagttc aggatgtgtg
     1681 gtcttctctt cgactaccca gtggtcttgg ctgttcaacc gtactttctc cagaaggatc
     1741 agctcaattt gctgctcaga tatttgggtt aagcaaccat ttggtatgga gcaaactgcg
     1801 agcaagcatt ttgaacacat ggatttcctt gaagcaggct gacaagaaaa tcagagaatg
     1861 taatttagga attctgcagt cgacggtacc gcgggcccgg gatccaccgg tcgccaccat
     1921 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1981 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2041 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2101 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2161 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2221 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2281 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2341 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2401 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2461 ccactaccag cagaacaccc ccatcggcga cggtcccgtg ctgctgcccg acaaccacta
     2521 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2581 gaaggagcgc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     2641 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2701 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2761 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2821 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2881 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2941 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3001 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3061 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3121 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3181 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3241 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3301 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3361 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3421 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3481 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3601 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3661 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3721 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3781 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3841 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3901 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3961 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4021 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4081 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4141 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4201 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4261 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4321 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4381 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4441 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4501 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4561 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4621 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4681 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4741 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4801 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4861 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4921 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4981 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5041 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5101 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5161 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5221 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5281 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5341 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5401 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5461 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5521 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5581 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5641 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5701 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5761 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgaacgtcga
     5821 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5881 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5941 gattctgtgg ataaccgtat taccgccatg cattagt
//
