LOCUS       sequence-204159-        6165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ADSL-EGFP fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6165)
  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 6165)
  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: pADSL-EGFP
FEATURES             Location/Qualifiers
     source          1..6165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        46..349
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        350..553
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     504..524
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2090..2099
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2096..2098,2099..2101,2102..2815)
                     /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(2141..2162)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2402..2421)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2938..3059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2975..2994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3029..3048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3066..3521)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3203..3224)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3415..3434
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3548..3652
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3654..4011
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3654..3845
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3677..3697)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3862..3997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4046..4840
                     /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             4046..4840
                     /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(4100..4119)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4710..4729
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5028..5047)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5072..5119
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5448..6036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5937..5956
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
       61 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      121 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      181 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      241 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      301 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      361 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      421 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      481 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      541 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
      601 gagctcaagc ttcgaattca tggcggctgg aggcgatcat ggttcgcccg acagctaccg
      661 ctcacctctt gcctcccgct atgccagccc ggagatgtgc ttcgtgttta gcgacaggta
      721 taaattccgg acatggcggc agctgtggct gtggctggcg gaggccgagc agacattggg
      781 tttgcctatc acagatgaac aaatccagga gatgaaatca aacctggaga acatcgactt
      841 caagatggca gctgaggaag agaaacgttt acgacatgat gtgatggctc acgtgcacac
      901 atttggccac tgctgtccaa aagctgcagg cattattcac cttggtgcta cttcttgcta
      961 tgttggagac aatactgact tgattattct tagaaatgca cttgacctgc ttttgccaaa
     1021 gcttgccaga gtgatctctc ggcttgccga ctttgctaag gaacgagcca gtctacccac
     1081 attaggtttc acacatttcc agcctgcaca gctgaccaca gttgggaaac gttgctgtct
     1141 ttggattcag gatctttgca tggatctcca gaacttgaag cgtgtccgag atgacctgcg
     1201 cttccgggga gtaaagggta ccactggcac tcaggccagt ttcctgcagc tctttgaggg
     1261 agatgaccat aaggtagagc agcttgacaa gatggtgaca gaaaaggcag gatttaagag
     1321 agctttcatc atcacagggc agacatatac acgaaaagtg gatattgaag tactgtctgt
     1381 gctggctagc ttgggggcat cagtgcacaa gatttgcacc gacatacgcc tcctggcaaa
     1441 cctcaaggag atggaggaac cctttgaaaa acagcagatt ggctcaagtg cgatgccata
     1501 taagcggaat cccatgcgtt cagaacgttg ctgcagtctt gcccgccacc tgatgaccct
     1561 tgtcatggac ccgctacaga cagcatctgt ccagtggttt gaacgcacac tggatgatag
     1621 tgccaaccga cggatctgtt tggccgaggc atttcttacc gcagatacta tattgaatac
     1681 gctgcagaac atttctgaag gattggtcgt gtaccccaaa gtaattgaac ggcgcattcg
     1741 gcaagagctg cctttcatgg ccacagagaa catcatcatg gccatggtca aagctggagg
     1801 tagccgccag gattgccatg agaaaatcag agtgctttct cagcaggcag cttctgtggt
     1861 taagcaggaa gggggtgaca atgacctcat agagcgtatc caggttgatg cctacttcag
     1921 tcccattcac tcccagttgg atcatttact ggatccttct tctttcactg gtcgtgcctc
     1981 ccagcaggtg cagagattct tagaagagga ggtgtatccc ctgttaaaac catatgaaag
     2041 cgtgatgaag gtgaaagcag aattatgtct ggcccgggat ccaccggtcg ccaccatggt
     2101 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2161 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2221 gctgaccctg aagttcatct gcaccaccgg aaagctgccc gtgccctggc ccaccctcgt
     2281 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2341 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2401 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2461 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2521 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2581 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2641 ctaccagcag aacaccccca tcggcgacgg tcccgtgctg ctgcccgaca accactacct
     2701 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgaa
     2761 ggagcgcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     2821 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2881 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2941 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3001 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3061 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3121 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3181 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3241 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3301 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3361 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3421 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3481 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3541 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3601 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3661 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3781 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3841 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3901 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3961 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4021 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4081 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4141 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4201 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4261 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4321 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4381 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4441 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4501 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4561 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4621 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4681 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4741 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4801 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4861 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4921 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4981 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5041 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5101 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5161 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5221 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5281 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5341 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5401 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5461 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5521 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5581 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5641 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5701 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5761 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5821 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5881 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5941 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6001 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6061 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6121 tctgtggata accgtattac cgccatgcat tagttattaa tagta
//
