LOCUS       sequence-386961-        6743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human wild-type PTPN22 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-WT-PTPN22
FEATURES             Location/Qualifiers
     source          1..6743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        615..633
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      668..677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             677..700
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3239..3258)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3240..3258)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3532..3653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3569..3588)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3623..3642
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3660..4115)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3797..3818)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4009..4028
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4142..4246
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4248..4605
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4248..4439
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4271..4291)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4456..4591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4640..5434
                     /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             4640..5434
                     /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(4694..4713)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5304..5323
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5622..5641)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5666..5713
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6042..6630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6531..6550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgat
      601 tacgccaagc tcgaaattaa ccctcactaa agggaacaaa agctggagct ccaccgcggt
      661 ggcggccgcc accatggatt acaaggatga cgacgataag agcccgggcg gatccatgga
      721 ccaaagagaa attctgcaga agttcctgga tgaggcccaa agcaagaaaa ttactaaaga
      781 ggagtttgcc aatgaatttc tgaagctgaa aaggcaatct accaagtaca aggcagacaa
      841 aacctatcct acaactgtgg ctgagaagcc caagaatatc aagaaaaaca gatataagga
      901 tattttgccc tatgattata gccgggtaga actatccctg ataacctctg atgaggattc
      961 cagctacatc aatgccaact tcattaaggg agtttatgga cccaaggctt atattgccac
     1021 ccagggtcct ttatctacaa ccctcctgga cttctggagg atgatttggg aatatagtgt
     1081 ccttatcatt gttatggcat gcatggagta tgaaatggga aagaaaaagt gtgagcgcta
     1141 ctgggctgag ccaggagaga tgcagctgga atttggccct ttctctgtat cctgtgaagc
     1201 tgaaaaaagg aaatctgatt atataatcag gactctaaaa gttaagttca atagtgaaac
     1261 tcgaactatc taccagtttc attacaagaa ttggccagac catgatgtac cttcatctat
     1321 agaccctatt cttgagctca tctgggatgt acgttgttac caagaggatg acagtgttcc
     1381 catatgcatt cactgcagtg ctggctgtgg aaggactggt gttatttgtg ctattgatta
     1441 tacatggatg ttgctaaaag atgggataat tcctgagaac ttcagtgttt tcagtttgat
     1501 ccgggaaatg cggacacaga ggccttcatt agttcaaacg caggaacaat atgaactggt
     1561 ctacaatgct gtattagaac tatttaagag acagatggat gttatcagag ataaacattc
     1621 tggaacagag agtcaagcaa agcattgtat tcctgagaaa aatcacactc tccaagcaga
     1681 ctcttattct cctaatttac caaaaagtac cacaaaagca gcaaaaatga tgaaccaaca
     1741 aaggacaaaa atggaaatca aagaatcttc ttcctttgac tttaggactt ctgaaataag
     1801 tgcaaaagaa gagctagttt tgcaccctgc taaatcaagc acttcttttg actttctgga
     1861 gctaaattac agttttgaca aaaatgctga cacaaccatg aaatggcaga caaaggcatt
     1921 tccaatagtt ggggagcctc ttcagaagca tcaaagtttg gatttgggct ctcttttgtt
     1981 tgagggatgt tctaattcta aacctgtaaa tgcagcagga agatatttta attcaaaggt
     2041 gccaataaca cggaccaaat caactccttt tgaattgata cagcagagag aaaccaagga
     2101 ggtggacagc aaggaaaact tttcttattt ggaatctcaa ccacatgatt cttgttttgt
     2161 agagatgcag gctcaaaaag taatgcatgt ttcttcagca gaactgaatt attcactgcc
     2221 atatgactct aaacaccaaa tacgtaatgc ctctaatgta aagcaccatg actctagtgc
     2281 tcttggtgta tattcttaca tacctttagt ggaaaatcct tatttttcat catggcctcc
     2341 aagtggtacc agttctaaga tgtctcttga tttacctgag aagcaagatg gaactgtttt
     2401 tccttcttct ctgttgccaa catcctctac atccctcttc tcttattaca attcacatga
     2461 ttctttatca ctgaattctc caaccaatat ttcctcacta ttgaaccagg agtcagctgt
     2521 actagcaact gctccaagga tagatgatga aatcccccct ccacttcctg tacggacacc
     2581 tgaatcattt attgtggttg aggaagctgg agaattctca ccaaatgttc ccaaatcctt
     2641 atcctcagct gtgaaggtaa aaattggaac atcactggaa tggggtggaa catctgaacc
     2701 aaagaaattt gatgactctg tgatacttag accaagcaag agtgtaaaac tccgaagtcc
     2761 taaatcagaa ctacatcaag atcgttcttc tcccccacct cctctcccag aaagaactct
     2821 agagtccttc tttcttgccg atgaagattg tatgcaggcc caatctatag aaacatattc
     2881 tactagctat cctgacacca tggaaaattc aacatcttca aaacagacac tgaagactcc
     2941 tggaaaaagt ttcacaagga gtaagagttt gaaaattttg cgaaacatga aaaagagtat
     3001 ctgtaattct tgcccaccaa acaagcctgc agaatctgtt cagtcaaata actccagctc
     3061 atttctgaat tttggttttg caaaccgttt ttcaaaaccc aaaggaccaa ggaatccacc
     3121 accaacttgg aatatttaat aaaactccag atttataata atatgggctg caagtacacc
     3181 tcgagggggg gcccggtacc ttaattaatt aaggtaccag gtaagtgtac ccaattcgcc
     3241 ctatagtgag tcgtattaca attcactcga tcgcccttcc caacagttgc gcagcctgaa
     3301 tggcgaatgg agatccaatt tttaagtgta taatgtgtta aactactgat tctaattgtt
     3361 tgtgtatttt agattcacag tcccaaggct catttcaggc ccctcagtcc tcacagtctg
     3421 ttcatgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3481 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3541 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3601 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     3661 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3721 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3781 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3841 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3901 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3961 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     4021 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4081 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     4141 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4201 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagaatcctg aggcggaaag
     4261 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4321 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4381 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4441 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4501 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4561 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4621 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4681 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4741 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4801 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4861 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4921 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4981 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5041 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5101 tcaggatgat ctggacgaag aacatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5161 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5221 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5281 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag aacttggcgg
     5341 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5401 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5461 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5521 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5581 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5641 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5701 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5761 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5821 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5881 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5941 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6001 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6061 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6121 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6181 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6241 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6301 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6361 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6421 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6481 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6541 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6601 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6661 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6721 gataaccgta ttaccgccat gca
//
