LOCUS       sequence-245747-        7028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human ADSL protein with a C-terminal 2XStrep affinity tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7028)
  AUTHORS   Liu C, Knudsen GM, Pedley AM, He J, Johnson JL, Yaron TM, Cantley 
            LC, Benkovic SJ
  TITLE     Mapping Post-Translational Modifications of de Novo Purine 
            Biosynthetic Enzymes: Implications for Pathway Regulation.
  JOURNAL   J Proteome Res. 2019 May 3;18(5):2078-2087. doi: 
            10.1021/acs.jproteome.8b00969. Epub 2019 Apr 18.
  PUBMED    30964683
REFERENCE   2  (bases 1 to 7028)
  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: 
            "10.1021/acs.jproteome.8b00969"; journalName: "J Proteome Res"; 
            date: "2019-05-3- 3"; volume: "18"; issue: "5"; pages: "2078-2087"
COMMENT     Sequence Label: pADSL-2XStrep
FEATURES             Location/Qualifiers
     source          1..7028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2592..2597,2598..2621,2622..2657,2658..2681)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    2732..2853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2743..2762)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2797..2816
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3039..3494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3126..3145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3336..3357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3611..3968
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3611..3802
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3634..3654)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3819..3954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4019..4813
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4073..4092)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4683..4702
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4877..4925
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4966..4985)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5085..5107
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5145..5163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5231..5335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5336..5404,5405..6196)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5554..5573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6367..6955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6856..6875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagctca agcttcgaat tcatggcggc tggaggcgat catggttcgc
     1141 ccgacagcta ccgctcacct cttgcctccc gctatgccag cccggagatg tgcttcgtgt
     1201 ttagcgacag gtataaattc cggacatggc ggcagctgtg gctgtggctg gcggaggccg
     1261 agcagacatt gggtttgcct atcacagatg aacaaatcca ggagatgaaa tcaaacctgg
     1321 agaacatcga cttcaagatg gcagctgagg aagagaaacg tttacgacat gatgtgatgg
     1381 ctcacgtgca cacatttggc cactgctgtc caaaagctgc aggcattatt caccttggtg
     1441 ctacttcttg ctatgttgga gacaatactg acttgattat tcttagaaat gcacttgacc
     1501 tgcttttgcc aaagcttgcc agagtgatct ctcggcttgc cgactttgct aaggaacgag
     1561 ccagtctacc cacattaggt ttcacacatt tccagcctgc acagctgacc acagttggga
     1621 aacgttgctg tctttggatt caggatcttt gcatggatct ccagaacttg aagcgtgtcc
     1681 gagatgacct gcgcttccgg ggagtaaagg gtaccactgg cactcaggcc agtttcctgc
     1741 agctctttga gggagatgac cataaggtag agcagcttga caagatggtg acagaaaagg
     1801 caggatttaa gagagctttc atcatcacag ggcagacata tacacgaaaa gtggatattg
     1861 aagtactgtc tgtgctggct agcttggggg catcagtgca caagatttgc accgacatac
     1921 gcctcctggc aaacctcaag gagatggagg aaccctttga aaaacagcag attggctcaa
     1981 gtgcgatgcc atataagcgg aatcccatgc gttcagaacg ttgctgcagt cttgcccgcc
     2041 acctgatgac ccttgtcatg gacccgctac agacagcatc tgtccagtgg tttgaacgca
     2101 cactggatga tagtgccaac cgacggatct gtttggccga ggcatttctt accgcagata
     2161 ctatattgaa tacgctgcag aacatttctg aaggattggt cgtgtacccc aaagtaattg
     2221 aacggcgcat tcggcaagag ctgcctttca tggccacaga gaacatcatc atggccatgg
     2281 tcaaagctgg aggtagccgc caggattgcc atgagaaaat cagagtgctt tctcagcagg
     2341 cagcttctgt ggttaagcag gaagggggtg acaatgacct catagagcgt atccaggttg
     2401 atgcctactt cagtcccatt cactcccagt tggatcattt actggatcct tcttctttca
     2461 ctggtcgtgc ctcccagcag gtgcagagat tcttagaaga ggaggtgtat cccctgttaa
     2521 aaccatatga aagcgtgatg aaggtgaaag cagaattatg tctggcccgg gatccaccgg
     2581 tcgccaccgg gagcgcttgg agccacccgc agttcgaaaa aggtggaggt tctggcggtg
     2641 gatcgggagg ttcagcgtgg agccacccgc agttcgagaa ataaagcggc cgcttccctt
     2701 tagtgagggt taatgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     2761 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2821 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2881 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     2941 ggtaaaatcc gataaggatc gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc
     3001 gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt
     3061 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3121 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3181 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3241 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3301 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3361 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3421 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3481 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3541 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     3601 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3721 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3781 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3841 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3901 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3961 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta gagccaccat
     4021 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4081 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4141 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4201 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4261 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4321 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4381 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4441 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4501 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4561 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4621 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4681 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4741 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4801 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4861 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     4921 gtgtgaatcg atagcgataa ggatccgcgt atggtgcact ctcagtacaa tctgctctga
     4981 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5041 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5101 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5161 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5221 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5281 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5341 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5401 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5461 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5521 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5581 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5641 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5701 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5761 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5821 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5881 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5941 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6001 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6061 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6121 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6181 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6241 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6301 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6361 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6421 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6481 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6541 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6601 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6661 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6721 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6781 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6841 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6901 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6961 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg
     7021 acagatct
//
