LOCUS       sequence-389527-        6364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (exon 1-17 without the
            P1 and P2 domains) with FLAG tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6364)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 6364)
  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-deltaP1P2-PTPN22
FEATURES             Location/Qualifiers
     source          1..6364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        618..636
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             680..703
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2863..2882)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2864..2882)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3156..3277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3193..3212)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3247..3266
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3284..3739)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3421..3442)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3633..3652
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3766..3870
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3872..4229
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3872..4063
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3895..3915)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4080..4215
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4264..5058
                     /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             4264..5058
                     /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(4318..4337)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4928..4947
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5246..5265)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5290..5337
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5666..6254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6155..6174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gattacgcca agctcgaaat taaccctcac taaagggaac aaaagctgga gctccaccgc
      661 ggtggcggcc gccaccatgg attacaagga tgacgacgat aagagcccgg gcggatccat
      721 ggaccaaaga gaaattctgc agaagttcct ggatgaggcc caaagcaaga aaattactaa
      781 agaggagttt gccaatgaat ttctgaagct gaaaaggcaa tctaccaagt acaaggcaga
      841 caaaacctat cctacaactg tggctgagaa gcccaagaat atcaagaaaa acagatataa
      901 ggatattttg ccctatgatt atagccgggt agaactatcc ctgataacct ctgatgagga
      961 ttccagctac atcaatgcca acttcattaa gggagtttat ggacccaagg cttatattgc
     1021 cacccagggt cctttatcta caaccctcct ggacttctgg aggatgattt gggaatatag
     1081 tgtccttatc attgttatgg catgcatgga gtatgaaatg ggaaagaaaa agtgtgagcg
     1141 ctactgggct gagccaggag agatgcagct ggaatttggc cctttctctg tatcctgtga
     1201 agctgaaaaa aggaaatctg attatataat caggactcta aaagttaagt tcaatagtga
     1261 aactcgaact atctaccagt ttcattacaa gaattggcca gaccatgatg taccttcatc
     1321 tatagaccct attcttgagc tcatctggga tgtacgttgt taccaagagg atgacagtgt
     1381 tcccatatgc attcactgca gtgctggctg tggaaggact ggtgttattt gtgctattga
     1441 ttatacatgg atgttgctaa aagatgggat aattcctgag aacttcagtg ttttcagttt
     1501 gatccgggaa atgcggacac agaggccttc attagttcaa acgcaggaac aatatgaact
     1561 ggtctacaat gctgtattag aactatttaa gagacagatg gatgttatca gagataaaca
     1621 ttctggaaca gagagtcaag caaagcattg tattcctgag aaaaatcaca ctctccaagc
     1681 agactcttat tctcctaatt taccaaaaag taccacaaaa gcagcaaaaa tgatgaacca
     1741 acaaaggaca aaaatggaaa tcaaagaatc ttcttccttt gactttagga cttctgaaat
     1801 aagtgcaaaa gaagagctag ttttgcaccc tgctaaatca agcacttctt ttgactttct
     1861 ggagctaaat tacagttttg acaaaaatgc tgacacaacc atgaaatggc agacaaaggc
     1921 atttccaata gttggggagc ctcttcagaa gcatcaaagt ttggatttgg gctctctttt
     1981 gtttgaggga tgttctaatt ctaaacctgt aaatgcagca ggaagatatt ttaattcaaa
     2041 ggtgccaata acacggacca aatcaactcc ttttgaattg atacagcaga gagaaaccaa
     2101 ggaggtggac agcaaggaaa acttttctta tttggaatct caaccacatg attcttgttt
     2161 tgtagagatg caggctcaaa aagtaatgca tgtttcttca gcagaactga attattcact
     2221 gccatatgac tctaaacacc aaatacgtaa tgcctctaat gtaaagcacc atgactctag
     2281 tgctcttggt gtatattctt acataccttt agtggaaaat ccttattttt catcatggcc
     2341 tccaagtggt accagttcta agatgtctct tgatttacct gagaagcaag atggaactgt
     2401 ttttccttct tctctgttgc caacatcctc tacatccctc ttctcttatt acaattcaca
     2461 tgattcttta tcactgaatt ctccaaccaa tatttcctca ctattgaacc aggagtcagc
     2521 tgtactagca actgctccaa ggatagatga tgaaatcgaa tcatttattg tggttgagga
     2581 agctggagaa ttctcaccaa atgttcccaa atccttatcc tcagctgtga aggtaaaaat
     2641 tggaacatca ctggaatggg gtggaacatc tgaaccaaag aaatttgatg actctgtgat
     2701 acttagacca agcaagagtg taaaactccg aagtcctaaa tcagaactac atcaagatcg
     2761 ttcttctact ctagagtcct tctttcttgc cgatgaagat tagctcgagg gggggcccgg
     2821 taccttaatt aattaaggta ccaggtaagt gtacccaatt cgccctatag tgagtcgtat
     2881 tacaattcac tcgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggagatcc
     2941 aatttttaag tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc
     3001 acagtcccaa ggctcatttc aggcccctca gtcctcacag tctgttcatg atcataatca
     3061 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3121 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3181 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3241 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     3301 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3361 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3421 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3481 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3541 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3601 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3661 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3721 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3781 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3841 atgcttcaat aatattgaaa aaggaagaat cctgaggcgg aaagaaccag ctgtggaatg
     3901 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3961 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4021 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4081 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4141 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4201 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4261 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4321 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4381 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4441 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4501 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4561 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4621 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4681 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4741 gaagaacatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4801 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4861 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4921 gacatagcgt tggctacccg tgatattgct gaagaacttg gcggcgaatg ggctgaccgc
     4981 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5041 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5101 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5161 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5221 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5281 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5341 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5401 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5461 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5521 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5581 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5641 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5701 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5761 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5821 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5881 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5941 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6001 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6061 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6121 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6181 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6241 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6301 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6361 ccat
//
