LOCUS       sequence-245741-        6828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human PAICS protein with a C-terminal 2XStrep affinity 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  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 6828)
  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: pPAICS-2XStrep
FEATURES             Location/Qualifiers
     source          1..6828
                     /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(2390..2395,2396..2419,2420..2455,2456..2479)
                     /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    2532..2653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2543..2562)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2597..2616
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2839..3294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2926..2945)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3136..3157
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3411..3768
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3411..3602
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3434..3454)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3619..3754
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3819..4613
                     /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(3873..3892)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4483..4502
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4677..4725
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4766..4785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4885..4907
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4945..4963)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5031..5135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5136..5204,5205..5996)
                     /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(5354..5373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6167..6755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6656..6675
                     /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 ctcgagatgg cgacagctga ggtactgaac attggtaaaa aattatatga
     1141 gggtaaaaca aaagaagtct acgaattgtt agacagtcca ggaaaagtcc tcctgcagtc
     1201 caaggaccag attacagcag gaaatgcagc tagaaaaaac cacctggaag gaaaagctgc
     1261 aatctcaaat aaaatcacca gttgtatttt tcagttatta caggaagcag gtattaaaac
     1321 tgccttcacc agaaaatgtg gggagacagc tttcattgca ccgcagtgtg aaatgattcc
     1381 aattgaatgg gtttgcagaa gaatagcaac tggttctttt ctcaaaagaa atcctggtgt
     1441 caaggaagga tataagtttt acccacctaa agtggagttg tttttcaagg atgatgccaa
     1501 taatgaccca cagtggtctg aggaacagct gattgctgca aaattttgct ttgctggact
     1561 tcttataggc cagactgaag tggatatcat gagtcatgct acacaggcta tatttgaaat
     1621 actggagaaa tcctggttgc cccagaattg tacactggtt gatatgaaga ttgaatttgg
     1681 tgttgatgta accaccaaag aaattgttct tgctgatgtt attgacaatg attcctggag
     1741 actctggcca tcaggagatc gaagccaaca gaaagacaaa cagtcttatc gggacctcaa
     1801 agaagtaact cctgaagggc tccaaatggt aaagaaaaac tttgagtggg ttgcagagag
     1861 agtagagttg ctcttgaaat cagaaagtca gtgcagggtt gtggtgttga tgggctctac
     1921 ttctgatctt ggtcactgtg aaaaaatcaa gaaggcctgt ggaaattttg gcattccatg
     1981 tgaacttcga gtaacatctg cgcataaagg accagatgaa actctgagga ttaaagctga
     2041 gtatgaaggg gatggcattc ctactgtatt tgtggcagtg gcaggcagaa gtaatggttt
     2101 gggaccagtg atgtctggga acactgcata tccagttatc agctgtcctc ccctcacacc
     2161 agactgggga gttcaggatg tgtggtcttc tcttcgacta cccagtggtc ttggctgttc
     2221 aaccgtactc tctccagaag gatcagctca atttgctgct cagatatttg ggttaagcaa
     2281 ccatttggta tggagcaaac tgcgagcaag cattttgaac acatggattt ccttgaagca
     2341 ggctgacaag aaaatcagag aatgtaattt accaccggtc gccaccggga gcgcttggag
     2401 ccacccgcag ttcgaaaaag gtggaggttc tggcggtgga tcgggaggtt cagcgtggag
     2461 ccacccgcag ttcgagaaat aaagcgcggc cgcttccctt tagtgagggt taatgcttcg
     2521 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2581 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2641 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2701 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcc gataaggatc
     2761 gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2821 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2881 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2941 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3001 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3061 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3121 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3181 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3241 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     3301 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     3361 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     3421 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3481 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3601 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3661 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3721 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct
     3781 tctgacacaa cagtctcgaa cttaaggcta gagccaccat gattgaacaa gatggattgc
     3841 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3901 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3961 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4021 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4081 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4141 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4201 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4261 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4321 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4381 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4441 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4501 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4561 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4621 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata
     4681 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa
     4741 ggatccgcgt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     4801 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4861 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4921 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     4981 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5041 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5101 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5161 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5221 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5281 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5341 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5401 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5461 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5521 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5581 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5641 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5701 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5761 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5821 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5881 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5941 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6001 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6061 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6121 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6181 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6241 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6301 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6361 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6421 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6481 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6541 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6601 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6661 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6721 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6781 ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatct
//
