LOCUS       sequence-387201-        6391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (exon 1-17 without the
            P2 domain) with FLAG tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6391)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 6391)
  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-deltaP2-PTPN22
FEATURES             Location/Qualifiers
     source          1..6391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        73..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     531..551
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        626..644
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      679..688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             688..711
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2898..2917)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2899..2917)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3191..3312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3228..3247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3282..3301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3319..3774)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3456..3477)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3668..3687
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3801..3905
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3907..4264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3907..4098
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3930..3950)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4115..4250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4299..5093
                     /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             4299..5093
                     /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(4353..4372)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4963..4982
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5281..5300)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5325..5372
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5701..6289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6190..6209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
       61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      601 ccgctagcga ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggagc
      661 tccaccgcgg tggcggccgc caccatggat tacaaggatg acgacgataa gagcccgggc
      721 ggatccatgg accaaagaga aattctgcag aagttcctgg atgaggccca aagcaagaaa
      781 attactaaag aggagtttgc caatgaattt ctgaagctga aaaggcaatc taccaagtac
      841 aaggcagaca aaacctatcc tacaactgtg gctgagaagc ccaagaatat caagaaaaac
      901 agatataagg atattttgcc ctatgattat agccgggtag aactatccct gataacctct
      961 gatgaggatt ccagctacat caatgccaac ttcattaagg gagtttatgg acccaaggct
     1021 tatattgcca cccagggtcc tttatctaca accctcctgg acttctggag gatgatttgg
     1081 gaatatagtg tccttatcat tgttatggca tgcatggagt atgaaatggg aaagaaaaag
     1141 tgtgagcgct actgggctga gccaggagag atgcagctgg aatttggccc tttctctgta
     1201 tcctgtgaag ctgaaaaaag gaaatctgat tatataatca ggactctaaa agttaagttc
     1261 aatagtgaaa ctcgaactat ctaccagttt cattacaaga attggccaga ccatgatgta
     1321 ccttcatcta tagaccctat tcttgagctc atctgggatg tacgttgtta ccaagaggat
     1381 gacagtgttc ccatatgcat tcactgcagt gctggctgtg gaaggactgg tgttatttgt
     1441 gctattgatt atacatggat gttgctaaaa gatgggataa ttcctgagaa cttcagtgtt
     1501 ttcagtttga tccgggaaat gcggacacag aggccttcat tagttcaaac gcaggaacaa
     1561 tatgaactgg tctacaatgc tgtattagaa ctatttaaga gacagatgga tgttatcaga
     1621 gataaacatt ctggaacaga gagtcaagca aagcattgta ttcctgagaa aaatcacact
     1681 ctccaagcag actcttattc tcctaattta ccaaaaagta ccacaaaagc agcaaaaatg
     1741 atgaaccaac aaaggacaaa aatggaaatc aaagaatctt cttcctttga ctttaggact
     1801 tctgaaataa gtgcaaaaga agagctagtt ttgcaccctg ctaaatcaag cacttctttt
     1861 gactttctgg agctaaatta cagttttgac aaaaatgctg acacaaccat gaaatggcag
     1921 acaaaggcat ttccaatagt tggggagcct cttcagaagc atcaaagttt ggatttgggc
     1981 tctcttttgt ttgagggatg ttctaattct aaacctgtaa atgcagcagg aagatatttt
     2041 aattcaaagg tgccaataac acggaccaaa tcaactcctt ttgaattgat acagcagaga
     2101 gaaaccaagg aggtggacag caaggaaaac ttttcttatt tggaatctca accacatgat
     2161 tcttgttttg tagagatgca ggctcaaaaa gtaatgcatg tttcttcagc agaactgaat
     2221 tattcactgc catatgactc taaacaccaa atacgtaatg cctctaatgt aaagcaccat
     2281 gactctagtg ctcttggtgt atattcttac atacctttag tggaaaatcc ttatttttca
     2341 tcatggcctc caagtggtac cagttctaag atgtctcttg atttacctga gaagcaagat
     2401 ggaactgttt ttccttcttc tctgttgcca acatcctcta catccctctt ctcttattac
     2461 aattcacatg attctttatc actgaattct ccaaccaata tttcctcact attgaaccag
     2521 gagtcagctg tactagcaac tgctccaagg atagatgatg aaatcccccc tccacttcct
     2581 gtacggacac ctgaatcatt tattgtggtt gaggaagctg gagaattctc accaaatgtt
     2641 cccaaatcct tatcctcagc tgtgaaggta aaaattggaa catcactgga atggggtgga
     2701 acatctgaac caaagaaatt tgatgactct gtgatactta gaccaagcaa gagtgtaaaa
     2761 ctccgaagtc ctaaatcaga actacatcaa gatcgttctt ctactctaga gtccttcttt
     2821 cttgccgatg aagattagct cgaggggggg cccggtacct taattaatta aggtaccagg
     2881 taagtgtacc caattcgccc tatagtgagt cgtattacaa ttcactcgat cgcccttccc
     2941 aacagttgcg cagcctgaat ggcgaatgga gatccaattt ttaagtgtat aatgtgttaa
     3001 actactgatt ctaattgttt gtgtatttta gattcacagt cccaaggctc atttcaggcc
     3061 cctcagtcct cacagtctgt tcatgatcat aatcagccat accacatttg tagaggtttt
     3121 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3181 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3241 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3301 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3361 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3421 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3481 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3541 tatgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3601 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3661 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3721 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3781 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3841 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3901 agaatcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3961 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4021 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4081 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4141 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4201 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4261 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4321 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4381 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4441 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4501 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4561 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4621 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4681 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4741 tggaagccgg tcttgtcgat caggatgatc tggacgaaga acatcagggg ctcgcgccag
     4801 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4861 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4921 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4981 ttgctgaaga acttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5041 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5101 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5161 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5221 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5281 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5341 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5401 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5461 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5521 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5581 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5641 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5701 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5761 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5821 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5881 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5941 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6001 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6061 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6121 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6181 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6241 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6301 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6361 gttatcccct gattctgtgg ataaccgtat t
//
