LOCUS       sequence-387206-        6665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (lacking the P1, P2 
            and P4 domains) with FLAG tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6665)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 6665)
  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-deltaP1P2P4-PTPN22
FEATURES             Location/Qualifiers
     source          1..6665
                     /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(3166..3185)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3167..3185)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3459..3580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3496..3515)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3550..3569
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3587..4042)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3724..3745)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3936..3955
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4069..4173
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4175..4532
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4175..4366
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4198..4218)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4383..4518
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4567..5361
                     /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             4567..5361
                     /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(4621..4640)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5231..5250
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5549..5568)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5593..5640
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5969..6557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6458..6477
                     /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 aacttggaat
     3061 atttaataaa actccagatt tataataata tgggctgcaa gtacacctcg agggggggcc
     3121 cggtacctta attaattaag gtaccaggta agtgtaccca attcgcccta tagtgagtcg
     3181 tattacaatt cactcgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaga
     3241 tccaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga
     3301 ttcacagtcc caaggctcat ttcaggcccc tcagtcctca cagtctgttc atgatcataa
     3361 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3421 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3481 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3541 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3601 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3661 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3721 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3781 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3841 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3901 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3961 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4021 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4081 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4141 taaatgcttc aataatattg aaaaaggaag aatcctgagg cggaaagaac cagctgtgga
     4201 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4261 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4321 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4381 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4441 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4501 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4561 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4621 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4681 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4741 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4801 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4861 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4921 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4981 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5041 gacgaagaac atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5101 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5161 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5221 caggacatag cgttggctac ccgtgatatt gctgaagaac ttggcggcga atgggctgac
     5281 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5341 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5401 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5461 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5521 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5581 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5641 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5701 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5761 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5821 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5881 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5941 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6001 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6061 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6121 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6181 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6241 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6301 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6361 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6421 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6481 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6541 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6601 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6661 ccgcc
//
