LOCUS       sequence-386195-        6674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (lacking the P1, P3 
            and P4 domains) with FLAG tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6674)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 6674)
  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-deltaP1P3P4-PTPN22
FEATURES             Location/Qualifiers
     source          1..6674
                     /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(3175..3194)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3176..3194)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3468..3589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3505..3524)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3559..3578
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3596..4051)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3733..3754)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3945..3964
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4078..4182
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4184..4541
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4184..4375
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4207..4227)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4392..4527
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4576..5370
                     /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             4576..5370
                     /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(4630..4649)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5240..5259
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5558..5577)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5602..5649
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5978..6566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6467..6486
                     /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 cgcagaatct
     3001 gttcagtcaa ataactccag ctcatttctg aattttggtt ttgcaaaccg tttttcaaaa
     3061 acttggaata tttaataaaa ctccagattt ataataatat gggctgcaag tacacctcga
     3121 gggggggccc ggtaccttaa ttaattaagg taccaggtaa gtgtacccaa ttcgccctat
     3181 agtgagtcgt attacaattc actcgatcgc ccttcccaac agttgcgcag cctgaatggc
     3241 gaatggagat ccaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     3301 tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac agtctgttca
     3361 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     3421 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3481 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3541 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     3601 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3661 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3721 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3781 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3841 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3901 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3961 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4021 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     4081 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4141 taaccctgat aaatgcttca ataatattga aaaaggaaga atcctgaggc ggaaagaacc
     4201 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4261 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4381 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4441 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4501 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4561 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4621 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4681 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4741 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4801 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4861 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4921 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4981 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5041 gatgatctgg acgaagaaca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5101 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5161 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5221 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagaact tggcggcgaa
     5281 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5341 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5401 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5461 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5521 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5581 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5641 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5701 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5761 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5821 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5881 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5941 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6001 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6061 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6121 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6181 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6241 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6301 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6361 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6421 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6481 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6541 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6601 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6661 accgtattac cgcc
//
