LOCUS       sequence-454084-        6080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expressed human ADSS with an EGFP fluorescent protein tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6080)
  AUTHORS   Zhao H, Chiaro CR, Zhang L, Smith PB, Chan CY, Pedley AM, Pugh RJ, 
            French JB, Patterson AD, Benkovic SJ
  TITLE     Quantitative analysis of purine nucleotides indicates that 
            purinosomes increase de novo purine biosynthesis.
  JOURNAL   J Biol Chem. 2015 Mar 13;290(11):6705-13. doi: 
            10.1074/jbc.M114.628701. Epub 2015 Jan 20.
  PUBMED    25605736
REFERENCE   2  (bases 1 to 6080)
  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: 
            "10.1074/jbc.M114.628701"; journalName: "J Biol Chem"; date: 
            "2015-03-13- 13"; volume: "290"; issue: "11"; pages: "6705-13"
COMMENT     Sequence Label: pADSS-EGFP
FEATURES             Location/Qualifiers
     source          1..6080
                     /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      2020..2029
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2026..2028,2029..2031,2032..2745)
                     /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(2071..2092)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2332..2351)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2868..2989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2905..2924)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2959..2978
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2996..3451)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3133..3154)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3345..3364
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3478..3582
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3584..3941
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3584..3775
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3607..3627)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3792..3927
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3976..4770
                     /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             3976..4770
                     /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(4030..4049)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4640..4659
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4958..4977)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5002..5049
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5378..5966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5867..5886
                     /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 gctagcgtcc
      601 accatggcgt tcgccgagac ctacccggcg gcatcctccc tgcccaacgg cgattgcggc
      661 cgccccaggg cgcggcccgg aggaaaccgg gtgacggtgg tgctcggtgc gcagtggggc
      721 gacgaaggca aagggaaggt ggtggacctg ctggcgcagg acgccgacat cgtgtgccgc
      781 tgccagggag gaaataatgc tggccataca gttgttgtgg attctgtgga atatgatttt
      841 catctcttac ccagtggaat aattaatcca aatgtcactg cattcattgg aaatggtgtg
      901 gtaattcatc tacctggatt gtttgaagaa gcagagaaaa atgttcaaaa aggaaaagga
      961 ctagaaggct gggaaaaaag gcttattata tctgacagag ctcatattgt atttgatttt
     1021 catcaagcag ctgatggtat ccaggaacaa cagagacaag aacaagcagg aaaaaatttg
     1081 ggtacaacaa aaaagggcat tggcccagtt tattcgtcca aagctgctcg gagtggactc
     1141 aggatgtgcg accttgtttc tgactttgat ggcttctctg agaggtttaa agttctagct
     1201 aaccaataca aatctatata ccccactttg gaaatagaca ttgaaggtga attacaaaaa
     1261 ctcaagggtt atatggaaaa gattaaacca atggtgagag atggagttta ttttctatat
     1321 gaggccctac atggaccacc aaagaaaatc ttggtagaag gtgcaaatgc agcactatta
     1381 gatattgatt ttgggactta cccttttgta acctcttcaa attgtactgt tggaggtgtt
     1441 tgtactggtt tgggtatgcc acctcaaaat gttggagaag tgtatggagt tgtgaaagct
     1501 tatacaacta gagttggtat tggtgccttt cctacagagc aagacaatga aattggagaa
     1561 ttattacaaa caaggggtag agagtttggt gtaactactg gaaggaaaag aagatgtggc
     1621 tggttggacc tcgttttgct caaatatgct catatgatca atggatttac tgcgttggca
     1681 cttaccaagt tggatatttt ggacatgttt acggaaatca aagttggagt tgcttacaag
     1741 ttagatggtg aaatcatacc tcatatccca gcaaaccaag aagtcttaaa taaagttgaa
     1801 gttcaatata agactctccc aggatggaac acagacatat caaatgcaag ggcgtttaaa
     1861 gaactacctg ttaatgcaca aaactatgtt cgatttattg aagatgagct tcaaattcca
     1921 gttaagtgga ttggtgttgg taaatccaga gaatctatga ttcaactctt ttggcgaatt
     1981 ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg ccaccatggt gagcaagggc
     2041 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2101 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2161 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2221 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2281 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2341 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2401 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2461 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     2521 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2581 aacaccccca tcggcgacgg tcccgtgctg ctgcccgaca accactacct gagcacccag
     2641 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgaa ggagcgcgtg
     2701 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     2761 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2821 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2881 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2941 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     3001 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3061 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3121 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3181 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3241 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3301 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3361 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3421 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3481 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3541 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3601 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3661 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3721 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3781 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3841 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3901 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3961 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4021 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4081 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4141 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4201 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4261 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4321 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4381 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4441 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4501 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4561 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4621 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4681 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4741 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4801 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4861 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4921 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4981 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5041 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5101 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5161 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5221 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5281 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5341 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5401 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5461 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5521 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5581 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5641 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5701 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5761 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5821 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5881 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5941 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6001 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6061 accgtattac cgccatgcat
//
