LOCUS       sequence-190557-        8816 bp    DNA     circular SYN 14-APR-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 Apr 14, 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        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      4756..4765
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4762..4764,4765..4767,4768..5481)
                     /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(4807..4828)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5068..5087)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    5604..5725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5641..5660)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5695..5714
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5732..6187)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5869..5890)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6081..6100
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6214..6318
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6320..6677
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6320..6511
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6343..6363)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6528..6663
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6712..7506
                     /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             6712..7506
                     /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(6766..6785)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7376..7395
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7694..7713)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7738..7785
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8114..8702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8603..8622
                     /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 gctagcatgc
      601 gcgagtgcat cttccacgaa cctaattcat ctctccagca aaggacacat ctctccagca
      661 aaggacacct ctctccagca aaggacacct gcagagatgt ccccagtcct tcacttctat
      721 gttcgtccct ctggccatga gggggcagcc cctggacaca ctcggaggaa actgcaaggg
      781 aaactgccag agctgcaggg cgtcgagact gaactgtgct acaacgtgaa ctggacagct
      841 gaggccctcc ccagtgctga ggagacaaag aagctgatgt ggctgtttgg ttgcccctta
      901 ctgctggatg atgttgctcg ggagtcctgg ctccttcctg gctccaatga cctgctgctg
      961 gaggtcgggc ccaggctgaa cttctccacc ccaacatcca ccaacatcgt gtcagtgtgc
     1021 cgcgccactg ggctggggcc tgtggatcgt gtggagacca cccggcgcta ccggctctcg
     1081 tttgcccacc ccccgtcagc tgaggtggaa gccattgctc tggctaccct gcacgaccgg
     1141 atgacagagc agcacttccc ccatcccatc cagagtttct cccctgagag catgccggaa
     1201 cccctcaatg gccctatcaa tatactgggt gagggccggc ttgcgctgga gaaggccaac
     1261 caggagcttg gtctggcttt agactcttgg gacctagact tctacaccaa gcgcttccag
     1321 gagctacagc ggaacccgag cactgtggag gcctttgact tggcgcagtc caatagcgag
     1381 cacagccgac actggttctt caagggccag ctccacgtgg atgggcagaa gctggtgcac
     1441 tcactgtttg agtccatcat gagcacccag gaatcctcga accccaacaa cgtcctcaaa
     1501 ttctgtgata acagcagtgc aatccaggga aaggaagtcc gattcctacg gcctgaggac
     1561 cccacacggc caagccgctt ccagcaacag caagggctga gacatgttgt cttcacagca
     1621 gagactcaca actttcccac aggagtatgc ccctttagtg gtgcaaccac tggcacaggg
     1681 ggccggattc gagatgtcca gtgcacaggc cgcggggccc acgtggtggc tggcactgcc
     1741 ggctattgct ttggaaatct gcatattcca ggttacaatc tgccctggga ggatccaagc
     1801 ttccagtatc ctgggaattt tgcccggccc ctggaggttg ccattgaagc cagtaatgga
     1861 gcttctgact atggcaacaa gtttggggaa ccagtgctgg ctggcttcgc ccgctccttg
     1921 ggcctccagc tcccagacgg ccagcggcgt gagtggatca agcccatcat gtttagtggg
     1981 ggcattgggt ccatggaagc tgaccacata agcaaggagg ccccagagcc aggcatggaa
     2041 gttgtaaagg ttggaggtcc cgtctacagg attggagttg gaggtggagc tgcttcatct
     2101 gtgcaggtgc agggagataa caccagtgac ctggactttg gggctgtgca gcgaggagac
     2161 ccggagatgg aacagaagat gaaccgtgtg atcagggctt gtgtggaggc ccccaaggga
     2221 aaccccatct gcagccttca tgatcagggc gctggtggca atggcaatgt cctaaaagag
     2281 ctgagtgacc cagctggagc catcatttac accagccgct tccagcttgg ggacccaacc
     2341 ctgaatgccc tggaaatctg gggggctgag taccaggaat caaatgctct tctgctgagg
     2401 tcccccaacc gggacttcct gactcatgtc agtgcccgtg aacgttgccc ggcttgcttc
     2461 gtgggcacca tcactggaga ccggagaata gtgctggtgg acgatcggga gtgtcctgtc
     2521 agaagaaatg gccaggggga tgcccccccg acacccctgc caacccctgt ggacctggag
     2581 ctcgaatggg tgctgggcaa gatgcctcgg aaggagttct tcctgcagag gaagcccccc
     2641 atgctgcagc ctctggcctt gcccccaggg ctgagcgtgc accaggctct ggagagggtt
     2701 ctgaggctgc ccgccgtggc cagcaagcgc tacctcacca ataaggtgga ccgctccgtg
     2761 ggaggcctgg tggcccagca gcagtgcgtg gggcccctgc aaactcctct ggcagatgta
     2821 gcggttgtgg cactgagcca tgaggagctc ataggggctg ccacagcctt gggagaacag
     2881 ccagtcaaga gcctgctgga cccaaaagtc gccgcccggc tggccgtggc cgaagccctc
     2941 accaacctgg tgtttgctct ggtcactgac ctccgggatg tgaagtgtag cgggaactgg
     3001 atgtgggcag ccaagctccc aggggagggc gcagctttgg cggatgcctg tgaggctatg
     3061 gtggcagtga tggcagccct gggtgtggca gtggatggtg gcaaggactc cctcagcatg
     3121 gctgctcggg ttggcactga gaccgtgcgg gctcctgggt cactggtcat ctcagcctat
     3181 gccgtctgcc cagacatcac agccactgtg accccagacc tcaagcatcc tgaagggaga
     3241 ggccatctgc tctatgtggc tctgagccct gggcagcacc ggctcggggg cacagctctg
     3301 gcccagtgct tctcccagct tggggaacac cctccagacc tggaccttcc tgagaacttg
     3361 gtgcgggcct tcagcatcac tcaggggctg ctgaaagacc gcctcctctg ctcaggccac
     3421 gatgtcagtg acggaggcct cgtcacatgc ctgctggaga tggcctttgc tggaaattgc
     3481 gggctacagg tggatgtgcc tgtccccagg gttgatgtcc tgtctgtgct gttcgctgag
     3541 gagccaggcc tcgtgctgga ggtgcaggag ccagacctgg cccaggtgct gaagcgttac
     3601 cgggatgctg gcctccattg cctggagctg ggccacacag gcgaggccgg gccccacgcc
     3661 atggtccggg tgtcagtgaa cggggctgtg gttctggagg agcctgttgg ggagctgcga
     3721 gccctctggg aggagacgag tttccagctg gaccggctac aggcagagcc tcgctgtgtg
     3781 gcagaggagg aacggggcct gagggagcgg atggggccca gctattgcct gccccccacc
     3841 tttcccaaag cctccgtgcc ccgtgagcct ggtggtccca gcccccgagt cgccatcttg
     3901 cgagaggagg gcagtaatgg agaccgggag atggccgatg ccttccactt agctgggttt
     3961 gaggtatggg acgtgaccat gcaggacctc tgctctgggg caattgggct ggacactttc
     4021 cgtggcgtgg ccttcgtggg cggcttcagc tatgcagatg tcctgggctc tgccaaaggg
     4081 tgggcagctg ctgtgacctt tcatcccagg gctggggctg agctgaggcg cttccggaag
     4141 cggccagaca ccttcagcct gggcgtgtgt aatggctgtc aactgctggc tctgctcggc
     4201 tgggtgggag gcgaccccaa tgaggatgct gcagagatgg gccctgactc ccagccagcc
     4261 cggccaggcc ttctgctacg ccacaacctg tctgggcgct acgagtctcg ctgggccagc
     4321 gtgcgtgtgg ggcctgggcc agccctgatg ctgcgaggga tggagggcgc cgtgctgccc
     4381 gtgtggagtg cgcacgggga aggttacgta gcattttctt ctccggaact ccaagctcag
     4441 attgaggcca ggggcttggc tccactgcac tgggctgatg atgacgggaa ccccacagag
     4501 cagtaccctc tgaatcccaa tgggtcccca gggggcgtgg ctggcatctg ctcctgtgat
     4561 ggccgccacc tggctgtcat gcctcaccct gagcgggccg ttaggccttg gcagtgggca
     4621 tggcgacccc ctccatttga tactctgacc acctccccct ggctccagct ctttatcaat
     4681 gcccgaaact ggaccctgga agggagctgc ggaattctgc agtcgacggt accgcgggcc
     4741 cgggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     4801 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4861 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4921 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     4981 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     5041 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     5101 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     5161 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     5221 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     5281 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggtccc
     5341 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     5401 gagaagcgcg atcacatggt cctgaaggag cgcgtgaccg ccgccgggat cactctcggc
     5461 atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat
     5521 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5581 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5641 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5701 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     5761 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5821 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5881 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5941 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     6001 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     6061 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6121 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6181 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6241 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6301 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     6361 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6421 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6481 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6541 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6601 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6661 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     6721 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     6781 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     6841 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     6901 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     6961 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     7021 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     7081 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     7141 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     7201 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     7261 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     7321 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     7381 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     7441 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     7501 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     7561 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     7621 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     7681 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     7741 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     7801 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     7861 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     7921 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     7981 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8041 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8101 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8161 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8221 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     8281 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8341 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8401 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8461 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8521 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8581 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8641 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8701 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8761 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
