LOCUS       sequence-387205-        6701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (lacking the P1 and P3
            domains) with FLAG tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6701)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 6701)
  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-deltaP1P3-PTPN22
FEATURES             Location/Qualifiers
     source          1..6701
                     /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(3202..3221)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3203..3221)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3495..3616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3532..3551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3586..3605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3623..4078)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3760..3781)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3972..3991
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4105..4209
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4211..4568
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4211..4402
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4234..4254)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4419..4554
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4603..5397
                     /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             4603..5397
                     /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(4657..4676)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5267..5286
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5585..5604)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5629..5676
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6005..6593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6494..6513
                     /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 cccaaaggac caaggaatcc accaccaact tggaatattt aataaaactc cagatttata
     3121 ataatatggg ctgcaagtac acctcgaggg ggggcccggt accttaatta attaaggtac
     3181 caggtaagtg tacccaattc gccctatagt gagtcgtatt acaattcact cgatcgccct
     3241 tcccaacagt tgcgcagcct gaatggcgaa tggagatcca atttttaagt gtataatgtg
     3301 ttaaactact gattctaatt gtttgtgtat tttagattca cagtcccaag gctcatttca
     3361 ggcccctcag tcctcacagt ctgttcatga tcataatcag ccataccaca tttgtagagg
     3421 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3481 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3541 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3601 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3661 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3721 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3781 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3841 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3901 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3961 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4021 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4081 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4141 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4201 aggaagaatc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4261 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4321 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4381 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4441 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4501 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4561 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4621 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4681 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4741 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4801 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4861 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4921 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4981 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5041 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagaacatca ggggctcgcg
     5101 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5161 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5221 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5281 gatattgctg aagaacttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5341 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5401 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5461 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5521 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5581 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5641 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5701 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5761 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5821 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5881 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5941 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6001 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6061 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6661 ctgcgttatc ccctgattct gtggataacc gtattaccgc c
//
