LOCUS       sequence-193178-        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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1917..1926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1923..1925,1926..1928,1929..2642)
                     /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(1968..1989)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2229..2248)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2765..2886
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2802..2821)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2856..2875
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2893..3348)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3030..3051)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3242..3261
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3375..3479
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3481..3838
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3481..3672
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3504..3524)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3689..3824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3873..4667
                     /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             3873..4667
                     /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(3927..3946)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4537..4556
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4855..4874)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4899..4946
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5275..5863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5764..5783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 cgacagctga ggtactgaac attggtaaaa aattatatga gggtaaaaca aaagaagtct
      661 acgaattgtt agacagtcca ggaaaagtcc tcctgcagtc caaggaccag attacagcag
      721 gaaatgcagc tagaaaaaac cacctggaag gaaaagctgc aatctcaaat aaaatcacca
      781 gttgtatttt tcagttatta caggaagcag gtattaaaac tgccttcacc agaaaatgtg
      841 gggagacagc tttcattgca ccgcagtgtg aaatgattcc aattgaatgg gtttgcagaa
      901 gaatagcaac tggttctttt ctcaaaagaa atcctggtgt caaggaagga tataagtttt
      961 acccacctaa agtggagttg tttttcaagg atgatgccaa taatgaccca cagtggtctg
     1021 aggaacagct gattgctgca aaattttgct ttgctggact tcttataggc cagactgaag
     1081 tggatatcat gagtcatgct acacaggcta tatttgaaat actggagaaa tcctggttgc
     1141 cccagaattg tacactggtt gatatgaaga ttgaatttgg tgttgatgta accaccaaag
     1201 aaattgttct tgctgatgtt attgacaatg attcctggag actctggcca tcaggagatc
     1261 gaagccaaca gaaagacaaa cagtcttatc gggacctcaa agaagtaact cctgaagggc
     1321 tccaaatggt aaagaaaaac tttgagtggg ttgcagagag agtagagttg cttttgaaat
     1381 cagaaagtca gtgcagggtt gtagtgttga tgggctctac ttctgatctt ggtcactgtg
     1441 aaaaaatcaa gaaggcctgt ggaaattttg gcattccatg tgaacttcga gtaacatctg
     1501 cgcataaagg accagatgaa actctgagga ttaaagctga gtatgaaggg gatggcattc
     1561 ctactgtatt tgtggcagtg gcaggcagaa gtaatggttt gggaccagtg atgtctggga
     1621 acactgcata tccagttatc agctgtcctc ccctcacacc agactgggga gttcaggatg
     1681 tgtggtcttc tcttcgacta cccagtggtc ttggctgttc aaccgtactt tctccagaag
     1741 gatcagctca atttgctgct cagatatttg ggttaagcaa ccatttggta tggagcaaac
     1801 tgcgagcaag cattttgaac acatggattt ccttgaagca ggctgacaag aaaatcagag
     1861 aatgtaattt aggaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     1921 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1981 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2041 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2101 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     2161 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2221 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2281 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2341 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     2401 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     2461 ccgaccacta ccagcagaac acccccatcg gcgacggtcc cgtgctgctg cccgacaacc
     2521 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     2581 tcctgaagga gcgcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     2641 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2701 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2761 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2821 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2881 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2941 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3001 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3061 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3121 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3181 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3241 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3301 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3361 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3421 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3481 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3541 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3601 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3661 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3721 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3781 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3841 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3901 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3961 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4021 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4081 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4141 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4201 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4261 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4321 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4381 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4441 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4501 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4561 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4621 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4681 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4741 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4801 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4861 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4921 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4981 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5041 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5101 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5161 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5221 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5281 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5341 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5401 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5461 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5521 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5581 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5641 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5701 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5761 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5821 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5881 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5941 ccctgattct gtggataacc gtattaccgc catgcat
//
