LOCUS       sequence-387195-        6415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (exon 1-17) with FLAG 
            tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6415)
  AUTHORS   Ge Y, Onengut-Gumuscu S, Quinlan AR, Mackey AJ, Wright JA, Buckner 
            JH, Habib T, Rich SS, Concannon P
  TITLE     Targeted Deep Sequencing in Multiple-Affected Sibships of European 
            Ancestry Identifies Rare Deleterious Variants in PTPN22 That Confer 
            Risk for Type 1 Diabetes.
  JOURNAL   Diabetes. 2016 Mar;65(3):794-802. doi: 10.2337/db15-0322. Epub 2015 
            Dec 2.
  PUBMED    26631741
REFERENCE   2  (bases 1 to 6415)
  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: "10.2337/db15-0322";
            journalName: "Diabetes"; date: "2016-03"; volume: "65"; issue: "3"; 
            pages: "794-802"
COMMENT     Sequence Label: FLAG-Ex1-17-PTPN22
FEATURES             Location/Qualifiers
     source          1..6415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        71..374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        375..578
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     529..549
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        624..642
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      677..686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             686..709
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2920..2939)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2921..2939)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3213..3334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3250..3269)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3304..3323
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3341..3796)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3478..3499)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3690..3709
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3823..3927
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3929..4286
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3929..4120
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3952..3972)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4137..4272
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4321..5115
                     /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             4321..5115
                     /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(4375..4394)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4985..5004
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5303..5322)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5347..5394
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5723..6311
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6212..6231
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
       61 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      181 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      241 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      301 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      361 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      421 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      481 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      541 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      601 gctagcgatt acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggagctc
      661 caccgcggtg gcggccgcca ccatggatta caaggatgac gacgataaga gcccgggcgg
      721 atccatggac caaagagaaa ttctgcagaa gttcctggat gaggcccaaa gcaagaaaat
      781 tactaaagag gagtttgcca atgaatttct gaagctgaaa aggcaatcta ccaagtacaa
      841 ggcagacaaa acctatccta caactgtggc tgagaagccc aagaatatca agaaaaacag
      901 atataaggat attttgccct atgattatag ccgggtagaa ctatccctga taacctctga
      961 tgaggattcc agctacatca atgccaactt cattaaggga gtttatggac ccaaggctta
     1021 tattgccacc cagggtcctt tatctacaac cctcctggac ttctggagga tgatttggga
     1081 atatagtgtc cttatcattg ttatggcatg catggagtat gaaatgggaa agaaaaagtg
     1141 tgagcgctac tgggctgagc caggagagat gcagctggaa tttggccctt tctctgtatc
     1201 ctgtgaagct gaaaaaagga aatctgatta tataatcagg actctaaaag ttaagttcaa
     1261 tagtgaaact cgaactatct accagtttca ttacaagaat tggccagacc atgatgtacc
     1321 ttcatctata gaccctattc ttgagctcat ctgggatgta cgttgttacc aagaggatga
     1381 cagtgttccc atatgcattc actgcagtgc tggctgtgga aggactggtg ttatttgtgc
     1441 tattgattat acatggatgt tgctaaaaga tgggataatt cctgagaact tcagtgtttt
     1501 cagtttgatc cgggaaatgc ggacacagag gccttcatta gttcaaacgc aggaacaata
     1561 tgaactggtc tacaatgctg tattagaact atttaagaga cagatggatg ttatcagaga
     1621 taaacattct ggaacagaga gtcaagcaaa gcattgtatt cctgagaaaa atcacactct
     1681 ccaagcagac tcttattctc ctaatttacc aaaaagtacc acaaaagcag caaaaatgat
     1741 gaaccaacaa aggacaaaaa tggaaatcaa agaatcttct tcctttgact ttaggacttc
     1801 tgaaataagt gcaaaagaag agctagtttt gcaccctgct aaatcaagca cttcttttga
     1861 ctttctggag ctaaattaca gttttgacaa aaatgctgac acaaccatga aatggcagac
     1921 aaaggcattt ccaatagttg gggagcctct tcagaagcat caaagtttgg atttgggctc
     1981 tcttttgttt gagggatgtt ctaattctaa acctgtaaat gcagcaggaa gatattttaa
     2041 ttcaaaggtg ccaataacac ggaccaaatc aactcctttt gaattgatac agcagagaga
     2101 aaccaaggag gtggacagca aggaaaactt ttcttatttg gaatctcaac cacatgattc
     2161 ttgttttgta gagatgcagg ctcaaaaagt aatgcatgtt tcttcagcag aactgaatta
     2221 ttcactgcca tatgactcta aacaccaaat acgtaatgcc tctaatgtaa agcaccatga
     2281 ctctagtgct cttggtgtat attcttacat acctttagtg gaaaatcctt atttttcatc
     2341 atggcctcca agtggtacca gttctaagat gtctcttgat ttacctgaga agcaagatgg
     2401 aactgttttt ccttcttctc tgttgccaac atcctctaca tccctcttct cttattacaa
     2461 ttcacatgat tctttatcac tgaattctcc aaccaatatt tcctcactat tgaaccagga
     2521 gtcagctgta ctagcaactg ctccaaggat agatgatgaa atcccccctc cacttcctgt
     2581 acggacacct gaatcattta ttgtggttga ggaagctgga gaattctcac caaatgttcc
     2641 caaatcctta tcctcagctg tgaaggtaaa aattggaaca tcactggaat ggggtggaac
     2701 atctgaacca aagaaatttg atgactctgt gatacttaga ccaagcaaga gtgtaaaact
     2761 ccgaagtcct aaatcagaac tacatcaaga tcgttcttct cccccacctc ctctcccaga
     2821 aagaactcta gagtccttct ttcttgccga tgaagattag ctcgaggggg ggcccggtac
     2881 cttaattaat taaggtacca ggtaagtgta cccaattcgc cctatagtga gtcgtattac
     2941 aattcactcg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gagatccaat
     3001 ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattcaca
     3061 gtcccaaggc tcatttcagg cccctcagtc ctcacagtct gttcatgatc ataatcagcc
     3121 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     3181 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     3241 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3301 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     3361 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3421 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3481 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3541 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3601 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3661 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3721 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3781 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3841 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3901 cttcaataat attgaaaaag gaagaatcct gaggcggaaa gaaccagctg tggaatgtgt
     3961 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4021 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4081 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4141 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4201 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4261 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4321 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4381 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4441 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4501 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4561 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4621 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4681 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4741 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4801 gaacatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4861 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4921 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4981 atagcgttgg ctacccgtga tattgctgaa gaacttggcg gcgaatgggc tgaccgcttc
     5041 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5101 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5161 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5221 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5281 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5341 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5401 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5461 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5521 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5581 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5641 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5701 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5761 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5821 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5881 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5941 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6001 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6061 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6121 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6181 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6241 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6301 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6361 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attac
//
