LOCUS       sequence-386193-        6692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (lacking the P1 and P2
            domains) with FLAG tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 6692)
  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-deltaP1P2-PTPN22
FEATURES             Location/Qualifiers
     source          1..6692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             682..705
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3193..3212)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3194..3212)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3486..3607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3523..3542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3577..3596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3614..4069)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3751..3772)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3963..3982
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4096..4200
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4202..4559
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4202..4393
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4225..4245)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4410..4545
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4594..5388
                     /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             4594..5388
                     /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(4648..4667)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5258..5277
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5576..5595)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5620..5667
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5996..6584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6485..6504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgccaccat ggattacaag gatgacgacg ataagagccc gggcggatcc
      721 atggaccaaa gagaaattct gcagaagttc ctggatgagg cccaaagcaa gaaaattact
      781 aaagaggagt ttgccaatga atttctgaag ctgaaaaggc aatctaccaa gtacaaggca
      841 gacaaaacct atcctacaac tgtggctgag aagcccaaga atatcaagaa aaacagatat
      901 aaggatattt tgccctatga ttatagccgg gtagaactat ccctgataac ctctgatgag
      961 gattccagct acatcaatgc caacttcatt aagggagttt atggacccaa ggcttatatt
     1021 gccacccagg gtcctttatc tacaaccctc ctggacttct ggaggatgat ttgggaatat
     1081 agtgtcctta tcattgttat ggcatgcatg gagtatgaaa tgggaaagaa aaagtgtgag
     1141 cgctactggg ctgagccagg agagatgcag ctggaatttg gccctttctc tgtatcctgt
     1201 gaagctgaaa aaaggaaatc tgattatata atcaggactc taaaagttaa gttcaatagt
     1261 gaaactcgaa ctatctacca gtttcattac aagaattggc cagaccatga tgtaccttca
     1321 tctatagacc ctattcttga gctcatctgg gatgtacgtt gttaccaaga ggatgacagt
     1381 gttcccatat gcattcactg cagtgctggc tgtggaagga ctggtgttat ttgtgctatt
     1441 gattatacat ggatgttgct aaaagatggg ataattcctg agaacttcag tgttttcagt
     1501 ttgatccggg aaatgcggac acagaggcct tcattagttc aaacgcagga acaatatgaa
     1561 ctggtctaca atgctgtatt agaactattt aagagacaga tggatgttat cagagataaa
     1621 cattctggaa cagagagtca agcaaagcat tgtattcctg agaaaaatca cactctccaa
     1681 gcagactctt attctcctaa tttaccaaaa agtaccacaa aagcagcaaa aatgatgaac
     1741 caacaaagga caaaaatgga aatcaaagaa tcttcttcct ttgactttag gacttctgaa
     1801 ataagtgcaa aagaagagct agttttgcac cctgctaaat caagcacttc ttttgacttt
     1861 ctggagctaa attacagttt tgacaaaaat gctgacacaa ccatgaaatg gcagacaaag
     1921 gcatttccaa tagttgggga gcctcttcag aagcatcaaa gtttggattt gggctctctt
     1981 ttgtttgagg gatgttctaa ttctaaacct gtaaatgcag caggaagata ttttaattca
     2041 aaggtgccaa taacacggac caaatcaact ccttttgaat tgatacagca gagagaaacc
     2101 aaggaggtgg acagcaagga aaacttttct tatttggaat ctcaaccaca tgattcttgt
     2161 tttgtagaga tgcaggctca aaaagtaatg catgtttctt cagcagaact gaattattca
     2221 ctgccatatg actctaaaca ccaaatacgt aatgcctcta atgtaaagca ccatgactct
     2281 agtgctcttg gtgtatattc ttacatacct ttagtggaaa atccttattt ttcatcatgg
     2341 cctccaagtg gtaccagttc taagatgtct cttgatttac ctgagaagca agatggaact
     2401 gtttttcctt cttctctgtt gccaacatcc tctacatccc tcttctctta ttacaattca
     2461 catgattctt tatcactgaa ttctccaacc aatatttcct cactattgaa ccaggagtca
     2521 gctgtactag caactgctcc aaggatagat gatgaaatcg aatcatttat tgtggttgag
     2581 gaagctggag aattctcacc aaatgttccc aaatccttat cctcagctgt gaaggtaaaa
     2641 attggaacat cactggaatg gggtggaaca tctgaaccaa agaaatttga tgactctgtg
     2701 atacttagac caagcaagag tgtaaaactc cgaagtccta aatcagaact acatcaagat
     2761 cgttcttcta ctctagagtc cttctttctt gccgatgaag attgtatgca ggcccaatct
     2821 atagaaacat attctactag ctatcctgac accatggaaa attcaacatc ttcaaaacag
     2881 acactgaaga ctcctggaaa aagtttcaca aggagtaaga gtttgaaaat tttgcgaaac
     2941 atgaaaaaga gtatctgtaa ttcttgccca ccaaacaagc ctgcagaatc tgttcagtca
     3001 aataactcca gctcatttct gaattttggt tttgcaaacc gtttttcaaa acccaaagga
     3061 ccaaggaatc caccaccaac ttggaatatt taataaaact ccagatttat aataatatgg
     3121 gctgcaagta cacctcgagg gggggcccgg taccttaatt aattaaggta ccaggtaagt
     3181 gtacccaatt cgccctatag tgagtcgtat tacaattcac tcgatcgccc ttcccaacag
     3241 ttgcgcagcc tgaatggcga atggagatcc aatttttaag tgtataatgt gttaaactac
     3301 tgattctaat tgtttgtgta ttttagattc acagtcccaa ggctcatttc aggcccctca
     3361 gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag gttttacttg
     3421 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3481 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3541 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3601 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3661 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3721 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3781 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3841 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3901 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3961 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4021 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4081 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4141 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagaat
     4201 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4261 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4321 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4381 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4441 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4501 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4561 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4621 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4681 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4741 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4801 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4861 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4921 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4981 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5041 gccggtcttg tcgatcagga tgatctggac gaagaacatc aggggctcgc gccagccgaa
     5101 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     5161 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5221 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5281 gaagaacttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5341 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5401 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5461 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5521 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5581 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5641 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5701 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5761 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5821 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5881 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5941 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6001 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6061 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6121 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6181 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6241 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6301 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6361 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6421 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6481 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6541 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6601 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6661 cccctgattc tgtggataac cgtattaccg cc
//
