LOCUS       sequence-204155-        8816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human PFAS-EGFP fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8816)
  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 8816)
  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: pFGAMS-EGFP
FEATURES             Location/Qualifiers
     source          1..8816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4757..4766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4763..4765,4766..4768,4769..5482)
                     /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(4808..4829)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5069..5088)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    5605..5726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5642..5661)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5696..5715
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5733..6188)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5870..5891)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6082..6101
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6215..6319
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6321..6678
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6321..6512
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6344..6364)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6529..6664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6713..7507
                     /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             6713..7507
                     /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(6767..6786)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7377..7396
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7695..7714)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7739..7786
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8115..8703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8604..8623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcatg
      601 cgcgagtgca tcttccacga acctaattca tctctccagc aaaggacaca tctctccagc
      661 aaaggacacc tctctccagc aaaggacacc tgcagagatg tccccagtcc ttcacttcta
      721 tgttcgtccc tctggccatg agggggcagc ccctggacac actcggagga aactgcaagg
      781 gaaactgcca gagctgcagg gcgtcgagac tgaactgtgc tacaacgtga actggacagc
      841 tgaggccctc cccagtgctg aggagacaaa gaagctgatg tggctgtttg gttgcccctt
      901 actgctggat gatgttgctc gggagtcctg gctccttcct ggctccaatg acctgctgct
      961 ggaggtcggg cccaggctga acttctccac cccaacatcc accaacatcg tgtcagtgtg
     1021 ccgcgccact gggctggggc ctgtggatcg tgtggagacc acccggcgct accggctctc
     1081 gtttgcccac cccccgtcag ctgaggtgga agccattgct ctggctaccc tgcacgaccg
     1141 gatgacagag cagcacttcc cccatcccat ccagagtttc tcccctgaga gcatgccgga
     1201 acccctcaat ggccctatca atatactggg tgagggccgg cttgcgctgg agaaggccaa
     1261 ccaggagctt ggtctggctt tagactcttg ggacctagac ttctacacca agcgcttcca
     1321 ggagctacag cggaacccga gcactgtgga ggcctttgac ttggcgcagt ccaatagcga
     1381 gcacagccga cactggttct tcaagggcca gctccacgtg gatgggcaga agctggtgca
     1441 ctcactgttt gagtccatca tgagcaccca ggaatcctcg aaccccaaca acgtcctcaa
     1501 attctgtgat aacagcagtg caatccaggg aaaggaagtc cgattcctac ggcctgagga
     1561 ccccacacgg ccaagccgct tccagcaaca gcaagggctg agacatgttg tcttcacagc
     1621 agagactcac aactttccca caggagtatg cccctttagt ggtgcaacca ctggcacagg
     1681 gggccggatt cgagatgtcc agtgcacagg ccgcggggcc cacgtggtgg ctggcactgc
     1741 cggctattgc tttggaaatc tgcatattcc aggttacaat ctgccctggg aggatccaag
     1801 cttccagtat cctgggaatt ttgcccggcc cctggaggtt gccattgaag ccagtaatgg
     1861 agcttctgac tatggcaaca agtttgggga accagtgctg gctggcttcg cccgctcctt
     1921 gggcctccag ctcccagacg gccagcggcg tgagtggatc aagcccatca tgtttagtgg
     1981 gggcattggg tccatggaag ctgaccacat aagcaaggag gccccagagc caggcatgga
     2041 agttgtaaag gttggaggtc ccgtctacag gattggagtt ggaggtggag ctgcttcatc
     2101 tgtgcaggtg cagggagata acaccagtga cctggacttt ggggctgtgc agcgaggaga
     2161 cccggagatg gaacagaaga tgaaccgtgt gatcagggct tgtgtggagg cccccaaggg
     2221 aaaccccatc tgcagccttc atgatcaggg cgctggtggc aatggcaatg tcctaaaaga
     2281 gctgagtgac ccagctggag ccatcattta caccagccgc ttccagcttg gggacccaac
     2341 cctgaatgcc ctggaaatct ggggggctga gtaccaggaa tcaaatgctc ttctgctgag
     2401 gtcccccaac cgggacttcc tgactcatgt cagtgcccgt gaacgttgcc cggcttgctt
     2461 cgtgggcacc atcactggag accggagaat agtgctggtg gacgatcggg agtgtcctgt
     2521 cagaagaaat ggccaggggg atgccccccc gacacccctg ccaacccctg tggacctgga
     2581 gctcgaatgg gtgctgggca agatgcctcg gaaggagttc ttcctgcaga ggaagccccc
     2641 catgctgcag cctctggcct tgcccccagg gctgagcgtg caccaggctc tggagagggt
     2701 tctgaggctg cccgccgtgg ccagcaagcg ctacctcacc aataaggtgg accgctccgt
     2761 gggaggcctg gtggcccagc agcagtgcgt ggggcccctg caaactcctc tggcagatgt
     2821 agcggttgtg gcactgagcc atgaggagct cataggggct gccacagcct tgggagaaca
     2881 gccagtcaag agcctgctgg acccaaaagt cgccgcccgg ctggccgtgg ccgaagccct
     2941 caccaacctg gtgtttgctc tggtcactga cctccgggat gtgaagtgta gcgggaactg
     3001 gatgtgggca gccaagctcc caggggaggg cgcagctttg gcggatgcct gtgaggctat
     3061 ggtggcagtg atggcagccc tgggtgtggc agtggatggt ggcaaggact ccctcagcat
     3121 ggctgctcgg gttggcactg agaccgtgcg ggctcctggg tcactggtca tctcagccta
     3181 tgccgtctgc ccagacatca cagccactgt gaccccagac ctcaagcatc ctgaagggag
     3241 aggccatctg ctctatgtgg ctctgagccc tgggcagcac cggctcgggg gcacagctct
     3301 ggcccagtgc ttctcccagc ttggggaaca ccctccagac ctggaccttc ctgagaactt
     3361 ggtgcgggcc ttcagcatca ctcaggggct gctgaaagac cgcctcctct gctcaggcca
     3421 cgatgtcagt gacggaggcc tcgtcacatg cctgctggag atggcctttg ctggaaattg
     3481 cgggctacag gtggatgtgc ctgtccccag ggttgatgtc ctgtctgtgc tgttcgctga
     3541 ggagccaggc ctcgtgctgg aggtgcagga gccagacctg gcccaggtgc tgaagcgtta
     3601 ccgggatgct ggcctccatt gcctggagct gggccacaca ggcgaggccg ggccccacgc
     3661 catggtccgg gtgtcagtga acggggctgt ggttctggag gagcctgttg gggagctgcg
     3721 agccctctgg gaggagacga gtttccagct ggaccggcta caggcagagc ctcgctgtgt
     3781 ggcagaggag gaacggggcc tgagggagcg gatggggccc agctattgcc tgccccccac
     3841 ctttcccaaa gcctccgtgc cccgtgagcc tggtggtccc agcccccgag tcgccatctt
     3901 gcgagaggag ggcagtaatg gagaccggga gatggccgat gccttccact tagctgggtt
     3961 tgaggtatgg gacgtgacca tgcaggacct ctgctctggg gcaattgggc tggacacttt
     4021 ccgtggcgtg gccttcgtgg gcggcttcag ctatgcagat gtcctgggct ctgccaaagg
     4081 gtgggcagct gctgtgacct ttcatcccag ggctggggct gagctgaggc gcttccggaa
     4141 gcggccagac accttcagcc tgggcgtgtg taatggctgt caactgctgg ctctgctcgg
     4201 ctgggtggga ggcgacccca atgaggatgc tgcagagatg ggccctgact cccagccagc
     4261 ccggccaggc cttctgctac gccacaacct gtctgggcgc tacgagtctc gctgggccag
     4321 cgtgcgtgtg gggcctgggc cagccctgat gctgcgaggg atggagggcg ccgtgctgcc
     4381 cgtgtggagt gcgcacgggg aaggttacgt agcattttct tctccggaac tccaagctca
     4441 gattgaggcc aggggcttgg ctccactgca ctgggctgat gatgacggga accccacaga
     4501 gcagtaccct ctgaatccca atgggtcccc agggggcgtg gctggcatct gctcctgtga
     4561 tggccgccac ctggctgtca tgcctcaccc tgagcgggcc gttaggcctt ggcagtgggc
     4621 atggcgaccc cctccatttg atactctgac cacctccccc tggctccagc tctttatcaa
     4681 tgcccgaaac tggaccctgg aagggagctg cggaattctg cagtcgacgg taccgcgggc
     4741 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     4801 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     4861 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     4921 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     4981 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     5041 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     5101 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     5161 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     5221 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     5281 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggtcc
     5341 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     5401 cgagaagcgc gatcacatgg tcctgaagga gcgcgtgacc gccgccggga tcactctcgg
     5461 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     5521 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5581 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5641 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5701 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     5761 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5821 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5881 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5941 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6001 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6061 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6121 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6181 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6241 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6301 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     6361 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6421 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6481 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6541 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6601 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6661 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     6721 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6781 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     6841 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     6901 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     6961 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     7021 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     7081 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     7141 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     7201 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     7261 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     7321 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     7381 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     7441 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     7501 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     7561 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     7621 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     7681 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     7741 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     7801 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     7861 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     7921 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     7981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8221 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgca
//
