LOCUS       sequence-387198-        6388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (exon 1-17 without the
            P1 domain) with FLAG tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6388)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 6388)
  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"
COMMENT     Sequence Label: FLAG-Ex1-17-deltaP1-PTPN22
FEATURES             Location/Qualifiers
     source          1..6388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..361
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        362..565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     516..536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        611..629
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      664..673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             673..696
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2880..2899)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2881..2899)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3173..3294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3210..3229)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3264..3283
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3301..3756)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3438..3459)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3650..3669
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3783..3887
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3889..4246
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3889..4080
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3912..3932)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4097..4232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4281..5075
                     /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             4281..5075
                     /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(4335..4354)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4945..4964
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5263..5282)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5307..5354
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5683..6271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6172..6191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
       61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      361 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      541 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgattacg
      601 ccaagctcga aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg
      661 gccgccacca tggattacaa ggatgacgac gataagagcc cgggcggatc catggaccaa
      721 agagaaattc tgcagaagtt cctggatgag gcccaaagca agaaaattac taaagaggag
      781 tttgccaatg aatttctgaa gctgaaaagg caatctacca agtacaaggc agacaaaacc
      841 tatcctacaa ctgtggctga gaagcccaag aatatcaaga aaaacagata taaggatatt
      901 ttgccctatg attatagccg ggtagaacta tccctgataa cctctgatga ggattccagc
      961 tacatcaatg ccaacttcat taagggagtt tatggaccca aggcttatat tgccacccag
     1021 ggtcctttat ctacaaccct cctggacttc tggaggatga tttgggaata tagtgtcctt
     1081 atcattgtta tggcatgcat ggagtatgaa atgggaaaga aaaagtgtga gcgctactgg
     1141 gctgagccag gagagatgca gctggaattt ggccctttct ctgtatcctg tgaagctgaa
     1201 aaaaggaaat ctgattatat aatcaggact ctaaaagtta agttcaatag tgaaactcga
     1261 actatctacc agtttcatta caagaattgg ccagaccatg atgtaccttc atctatagac
     1321 cctattcttg agctcatctg ggatgtacgt tgttaccaag aggatgacag tgttcccata
     1381 tgcattcact gcagtgctgg ctgtggaagg actggtgtta tttgtgctat tgattataca
     1441 tggatgttgc taaaagatgg gataattcct gagaacttca gtgttttcag tttgatccgg
     1501 gaaatgcgga cacagaggcc ttcattagtt caaacgcagg aacaatatga actggtctac
     1561 aatgctgtat tagaactatt taagagacag atggatgtta tcagagataa acattctgga
     1621 acagagagtc aagcaaagca ttgtattcct gagaaaaatc acactctcca agcagactct
     1681 tattctccta atttaccaaa aagtaccaca aaagcagcaa aaatgatgaa ccaacaaagg
     1741 acaaaaatgg aaatcaaaga atcttcttcc tttgacttta ggacttctga aataagtgca
     1801 aaagaagagc tagttttgca ccctgctaaa tcaagcactt cttttgactt tctggagcta
     1861 aattacagtt ttgacaaaaa tgctgacaca accatgaaat ggcagacaaa ggcatttcca
     1921 atagttgggg agcctcttca gaagcatcaa agtttggatt tgggctctct tttgtttgag
     1981 ggatgttcta attctaaacc tgtaaatgca gcaggaagat attttaattc aaaggtgcca
     2041 ataacacgga ccaaatcaac tccttttgaa ttgatacagc agagagaaac caaggaggtg
     2101 gacagcaagg aaaacttttc ttatttggaa tctcaaccac atgattcttg ttttgtagag
     2161 atgcaggctc aaaaagtaat gcatgtttct tcagcagaac tgaattattc actgccatat
     2221 gactctaaac accaaatacg taatgcctct aatgtaaagc accatgactc tagtgctctt
     2281 ggtgtatatt cttacatacc tttagtggaa aatccttatt tttcatcatg gcctccaagt
     2341 ggtaccagtt ctaagatgtc tcttgattta cctgagaagc aagatggaac tgtttttcct
     2401 tcttctctgt tgccaacatc ctctacatcc ctcttctctt attacaattc acatgattct
     2461 ttatcactga attctccaac caatatttcc tcactattga accaggagtc agctgtacta
     2521 gcaactgctc caaggataga tgatgaaatc gaatcattta ttgtggttga ggaagctgga
     2581 gaattctcac caaatgttcc caaatcctta tcctcagctg tgaaggtaaa aattggaaca
     2641 tcactggaat ggggtggaac atctgaacca aagaaatttg atgactctgt gatacttaga
     2701 ccaagcaaga gtgtaaaact ccgaagtcct aaatcagaac tacatcaaga tcgttcttct
     2761 cccccacctc ctctcccaga aagaactcta gagtccttct ttcttgccga tgaagattag
     2821 ctcgaggggg ggcccggtac cttaattaat taaggtacca ggtaagtgta cccaattcgc
     2881 cctatagtga gtcgtattac aattcactcg atcgcccttc ccaacagttg cgcagcctga
     2941 atggcgaatg gagatccaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt
     3001 ttgtgtattt tagattcaca gtcccaaggc tcatttcagg cccctcagtc ctcacagtct
     3061 gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3121 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3181 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3241 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     3301 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3361 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3421 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3481 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3541 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3601 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3661 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3721 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3781 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3841 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatcct gaggcggaaa
     3901 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3961 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4021 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4081 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4141 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4201 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4261 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4321 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4381 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4441 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4501 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4561 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4621 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4681 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4741 atcaggatga tctggacgaa gaacatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4801 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4861 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4921 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gaacttggcg
     4981 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5041 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5101 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5161 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5221 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5281 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5341 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5401 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5461 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5521 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5581 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5641 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5701 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5761 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5821 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5881 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5941 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6001 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6061 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6121 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6181 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6241 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6301 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6361 ggataaccgt attaccgcca tgcattag
//
