LOCUS       sequence-386962-        6743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a human PTPN22 mutant (1858T) with FLAG tag 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6743)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 6743)
  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-1858T-PTPN22
FEATURES             Location/Qualifiers
     source          1..6743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        49..352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        353..556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     507..527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        602..620
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      655..664
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             664..687
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3226..3245)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3227..3245)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3519..3640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3556..3575)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3610..3629
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3647..4102)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3784..3805)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3996..4015
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4129..4233
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4235..4592
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4235..4426
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4258..4278)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4443..4578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4627..5421
                     /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             4627..5421
                     /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(4681..4700)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5291..5310
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5609..5628)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5653..5700
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6029..6617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6518..6537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
       61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      541 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgattac gccaagctcg
      601 aaattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgccacc
      661 atggattaca aggatgacga cgataagagc ccgggcggat ccatggacca aagagaaatt
      721 ctgcagaagt tcctggatga ggcccaaagc aagaaaatta ctaaagagga gtttgccaat
      781 gaatttctga agctgaaaag gcaatctacc aagtacaagg cagacaaaac ctatcctaca
      841 actgtggctg agaagcccaa gaatatcaag aaaaacagat ataaggatat tttgccctat
      901 gattatagcc gggtagaact atccctgata acctctgatg aggattccag ctacatcaat
      961 gccaacttca ttaagggagt ttatggaccc aaggcttata ttgccaccca gggtccttta
     1021 tctacaaccc tcctggactt ctggaggatg atttgggaat atagtgtcct tatcattgtt
     1081 atggcatgca tggagtatga aatgggaaag aaaaagtgtg agcgctactg ggctgagcca
     1141 ggagagatgc agctggaatt tggccctttc tctgtatcct gtgaagctga aaaaaggaaa
     1201 tctgattata taatcaggac tctaaaagtt aagttcaata gtgaaactcg aactatctac
     1261 cagtttcatt acaagaattg gccagaccat gatgtacctt catctataga ccctattctt
     1321 gagctcatct gggatgtacg ttgttaccaa gaggatgaca gtgttcccat atgcattcac
     1381 tgcagtgctg gctgtggaag gactggtgtt atttgtgcta ttgattatac atggatgttg
     1441 ctaaaagatg ggataattcc tgagaacttc agtgttttca gtttgatccg ggaaatgcgg
     1501 acacagaggc cttcattagt tcaaacgcag gaacaatatg aactggtcta caatgctgta
     1561 ttagaactat ttaagagaca gatggatgtt atcagagata aacattctgg aacagagagt
     1621 caagcaaagc attgtattcc tgagaaaaat cacactctcc aagcagactc ttattctcct
     1681 aatttaccaa aaagtaccac aaaagcagca aaaatgatga accaacaaag gacaaaaatg
     1741 gaaatcaaag aatcttcttc ctttgacttt aggacttctg aaataagtgc aaaagaagag
     1801 ctagttttgc accctgctaa atcaagcact tcttttgact ttctggagct aaattacagt
     1861 tttgacaaaa atgctgacac aaccatgaaa tggcagacaa aggcatttcc aatagttggg
     1921 gagcctcttc agaagcatca aagtttggat ttgggctctc ttttgtttga gggatgttct
     1981 aattctaaac ctgtaaatgc agcaggaaga tattttaatt caaaggtgcc aataacacgg
     2041 accaaatcaa ctccttttga attgatacag cagagagaaa ccaaggaggt ggacagcaag
     2101 gaaaactttt cttatttgga atctcaacca catgattctt gttttgtaga gatgcaggct
     2161 caaaaagtaa tgcatgtttc ttcagcagaa ctgaattatt cactgccata tgactctaaa
     2221 caccaaatac gtaatgcctc taatgtaaag caccatgact ctagtgctct tggtgtatat
     2281 tcttacatac ctttagtgga aaatccttat ttttcatcat ggcctccaag tggtaccagt
     2341 tctaagatgt ctcttgattt acctgagaag caagatggaa ctgtttttcc ttcttctctg
     2401 ttgccaacat cctctacatc cctcttctct tattacaatt cacatgattc tttatcactg
     2461 aattctccaa ccaatatttc ctcactattg aaccaggagt cagctgtact agcaactgct
     2521 ccaaggatag atgatgaaat cccccctcca cttcctgtat ggacacctga atcatttatt
     2581 gtggttgagg aagctggaga attctcacca aatgttccca aatccttatc ctcagctgtg
     2641 aaggtaaaaa ttggaacatc actggaatgg ggtggaacat ctgaaccaaa gaaatttgat
     2701 gactctgtga tacttagacc aagcaagagt gtaaaactcc gaagtcctaa atcagaacta
     2761 catcaagatc gttcttctcc cccacctcct ctcccagaaa gaactctaga gtccttcttt
     2821 cttgccgatg aagattgtat gcaggcccaa tctatagaaa catattctac tagctatcct
     2881 gacaccatgg aaaattcaac atcttcaaaa cagacactga agactcctgg aaaaagtttc
     2941 acaaggagta agagtttgaa aattttgcga aacatgaaaa agagtatctg taattcttgc
     3001 ccaccaaaca agcctgcaga atctgttcag tcaaataact ccagctcatt tctgaatttt
     3061 ggttttgcaa accgtttttc aaaacccaaa ggaccaagga atccaccacc aacttggaat
     3121 atttaataaa actccagatt tataataata tgggctgcaa gtacacctcg agggggggcc
     3181 cggtacctta attaattaag gtaccaggta agtgtaccca attcgcccta tagtgagtcg
     3241 tattacaatt cactcgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaga
     3301 tccaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga
     3361 ttcacagtcc caaggctcat ttcaggcccc tcagtcctca cagtctgttc atgatcataa
     3421 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3481 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3601 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3661 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3721 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3781 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3841 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3901 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3961 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4021 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4081 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4141 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4201 taaatgcttc aataatattg aaaaaggaag aatcctgagg cggaaagaac cagctgtgga
     4261 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4321 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4381 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4441 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4501 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4561 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4621 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4681 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4741 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4801 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4861 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4921 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4981 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5041 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5101 gacgaagaac atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5161 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5221 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5281 caggacatag cgttggctac ccgtgatatt gctgaagaac ttggcggcga atgggctgac
     5341 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5401 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5461 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5521 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5581 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5641 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5701 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5761 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5821 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5881 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5941 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6001 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6061 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6121 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6181 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6241 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6301 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6361 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6421 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6481 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6541 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6601 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6661 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6721 ccgccatgca ttagttatta ata
//
