LOCUS       sequence-245750-        7329 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses human ATIC protein with an N-terminal 2XStrep affinity 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7329)
  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 7329)
  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: 
            "10.1021/acs.jproteome.8b00969"; journalName: "J Proteome Res"; 
            date: "2019-05-3- 3"; volume: "18"; issue: "5"; pages: "2078-2087"
COMMENT     Sequence Label: p2XStrep-ATIC
FEATURES             Location/Qualifiers
     source          1..7329
                     /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(1097..1102,1103..1126,1127..1162,1163..1186)
                     /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    3033..3154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3044..3063)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3098..3117
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3340..3795
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3427..3446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3637..3658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3912..4269
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3912..4103
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3935..3955)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4120..4255
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4320..5114
                     /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(4374..4393)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4984..5003
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5178..5226
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5267..5286)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5386..5408
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5446..5464)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5532..5636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5637..5705,5706..6497)
                     /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(5855..5874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6668..7256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7157..7176
                     /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 atggcaagcg cttggagcca cccgcagttc gaaaaaggtg gaggttctgg
     1141 cggtggatcg ggaggttcag cgtggagcca cccgcagttc gagaaatccg gaaggatccg
     1201 gtcagtgatg gctcccggcc agctcgcctt atttagtgtc tctgacaaaa ccggccttgt
     1261 ggaatttgca agaaacctga ccgctcttgg tttgaatctg gtcgcttccg gagggactgc
     1321 aaaagctctc agggatgctg gtctggcagt cagagatgtc tctgagttga cgggatttcc
     1381 tgaaatgttg gggggacgtg tgaaaacttt gcatcctgca gtccatgctg gaatcctagc
     1441 tcgtaatatt ccagaagata atgctgacat ggccagactt gatttcaatc ttataagagt
     1501 tgttgcctgc aatctctatc cctttgtaaa gacagtggct tctccaggtg taactgttga
     1561 ggaggctgtg gagcaaattg acattggtgg agtaacctta ctgagagctg cagccaaaaa
     1621 ccacgctcga gtgacagtgg tgtgtgaacc agaggactat gtggtggtgt ccacggagat
     1681 gcagagctcc gagagtaagg acacctcctt ggagactaga cgccagttag ccttgaaggc
     1741 attcactcat acggcacaat atgatgaagc aatttcagat tatttcagga aacagtacag
     1801 caaaggcgta tctcagatgc ccttgagata tggaatgaac ccacatcaga cccctgccca
     1861 gctgtacaca ctgcagccca agcttcccat cacagttcta aatggagccc ctggatttat
     1921 aaacttgtgc gatgctttga acgcctggca gctggtgaag gaactcaagg aggctttagg
     1981 tattccagcc gctgcctctt tcaaacatgt cagcccagca ggtgctgctg ttggaattcc
     2041 actcagtgaa gatgaggcca aagtctgcat ggtttatgat ctctataaaa ccctcacacc
     2101 catctcagcg gcgtatgcaa gagcaagagg ggctgatagg atgtcttcat ttggtgattt
     2161 tgttgcattg tccgatgttt gtgatgtacc aactgcaaaa attatttcca gagaagtatc
     2221 tgatggtata attgccccag gatatgaaga agaagccttg acaatacttt ccaaaaagaa
     2281 aaatggaaac tattgtgtcc ttcagatgga ccaatcttac aaaccagatg aaaatgaagt
     2341 tcgaactctc tttggtcttc atttaagcca gaagagaaat aatggtgtcg tcgacaagtc
     2401 attatttagc aatgttgtta ccaaaaataa agatttgcca gagtctgccc tccgagacct
     2461 catcgtagcc accattgctg tcaagtacac tcagtctaac tctgtgtgct acgccaagaa
     2521 cgggcaggtt atcggcattg gagcaggaca gcagtctcgt atacactgca ctcgccttgc
     2581 aggagataag gcaaactatt ggtggcttag acaccatcca caagtgcttt cgatgaagtt
     2641 taaaacagga gtgaagagag cagaaatctc caatgccatc gatcaatatg tgactggaac
     2701 cattggcgag gatgaagatt tgataaagtg gaaggcactg tttgaggaag tccctgagtt
     2761 actcactgag gcagagaaga aggaatgggt tgagaaactg actgaagttt ctatcagctc
     2821 tgatgccttc ttccctttcc gagataacgt agacagagct aaaaggagtg gtgtggcgta
     2881 cattgcggct ccctccggtt ctgctgctga caaagttgtg attgaggcct gcgacgaact
     2941 gggaatcatc ctcgctcata cgaaccttcg gctcttccac cactaagcgg ccgcttccct
     3001 ttagtgaggg ttaatgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     3061 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3121 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3181 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3241 tggtaaaatc cgataaggat cgatccgggc tggcgtaata gcgaagaggc ccgcaccgat
     3301 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     3361 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3421 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3481 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3541 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3601 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3661 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3721 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3781 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3841 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     3901 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3961 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4021 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4081 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4141 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4201 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4261 ttttgcaaaa agcttgattc ttctgacaca acagtctcga acttaaggct agagccacca
     4321 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4381 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4441 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4501 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4561 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4621 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4681 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4741 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4801 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4861 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4921 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4981 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5041 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5101 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5161 gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     5221 tgtgtgaatc gatagcgata agcatccgcg tatggtgcac tctcagtaca atctgctctg
     5281 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5341 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5401 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5461 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5521 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5581 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5641 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5701 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5761 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5821 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5881 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5941 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6001 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6061 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6121 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6181 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6241 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6301 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6361 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6421 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6481 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6541 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6601 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6661 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6721 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6781 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6841 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6901 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6961 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7021 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7081 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7141 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7201 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7261 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatggctc
     7321 gacagatct
//
