LOCUS       sequence-386191-        6689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (lacking the P1 and P4
            domains) with FLAG tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6689)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 6689)
  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-deltaP1P4-PTPN22
FEATURES             Location/Qualifiers
     source          1..6689
                     /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(3190..3209)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3191..3209)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3483..3604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3520..3539)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3574..3593
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3611..4066)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3748..3769)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3960..3979
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4093..4197
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4199..4556
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4199..4390
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4222..4242)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4407..4542
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4591..5385
                     /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             4591..5385
                     /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(4645..4664)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5255..5274
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5573..5592)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5617..5664
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5993..6581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6482..6501
                     /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 cgttcttctc ccccacctcc tctcccagaa agaactctag agtccttctt tcttgccgat
     2821 gaagattgta tgcaggccca atctatagaa acatattcta ctagctatcc tgacaccatg
     2881 gaaaattcaa catcttcaaa acagacactg aagactcctg gaaaaagttt cacaaggagt
     2941 aagagtttga aaattttgcg aaacatgaaa aagagtatct gtaattcttg cccaccaaac
     3001 aagcctgcag aatctgttca gtcaaataac tccagctcat ttctgaattt tggttttgca
     3061 aaccgttttt caaaaacttg gaatatttaa taaaactcca gatttataat aatatgggct
     3121 gcaagtacac ctcgaggggg ggcccggtac cttaattaat taaggtacca ggtaagtgta
     3181 cccaattcgc cctatagtga gtcgtattac aattcactcg atcgcccttc ccaacagttg
     3241 cgcagcctga atggcgaatg gagatccaat ttttaagtgt ataatgtgtt aaactactga
     3301 ttctaattgt ttgtgtattt tagattcaca gtcccaaggc tcatttcagg cccctcagtc
     3361 ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3421 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3481 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3541 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3601 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3661 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3721 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3781 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3841 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3901 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3961 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4021 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4081 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4141 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatcct
     4201 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4261 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4321 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4381 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4441 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4501 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4561 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4621 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4681 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4741 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4801 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4861 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4921 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4981 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5041 ggtcttgtcg atcaggatga tctggacgaa gaacatcagg ggctcgcgcc agccgaactg
     5101 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5161 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5221 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5281 gaacttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5341 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5401 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5461 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5521 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5581 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5641 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5701 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5761 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5821 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6121 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6661 ctgattctgt ggataaccgt attaccgcc
//
