LOCUS       sequence-338896-        7814 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses PTPIP51 FFAT-like motif (151-175) deletion mutant 
            (1-150_176-470) in human HeLa cells after transfection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7814)
  AUTHORS   Yeo HK, Park TH, Kim HY, Jang H, Lee J, Hwang GS, Ryu SE, Park SH, 
            Song HK, Ban HS, Yoon HJ, Lee BI
  TITLE     Phospholipid transfer function of PTPIP51 at mitochondria-associated
            ER membranes.
  JOURNAL   EMBO Rep. 2021 Jun 4;22(6):e51323. doi: 10.15252/embr.202051323. 
            Epub 2021 May 2.
  PUBMED    33938112
REFERENCE   2  (bases 1 to 7814)
  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"; journalName: "EMBO Rep.";
            date: "2021-06-4"; volume: "22(6):e51323. doi"; pages: " 
            10.15252/embr.202051323. Epub 2021 May 2"
COMMENT     Sequence Label: pCAG-FLAG-puro-PTPIP51(1-150_175-470)
FEATURES             Location/Qualifiers
     source          1..7814
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          188..1204
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1128..1150
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1212..1231
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1259..1268
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1268..1291
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    join(2659..3233,3234..3236,3237..3245)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2826..2843)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3053..3072
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3246..3845
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3246..3265)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3742..3762
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3926..3943)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3932..4156
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4203..4219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4203..4219)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4216..4238)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4227..4243
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4251..4257,4258..4275,4276..4281))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4296..4317
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4376..4571
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4422..4557
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4577..4711
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4779..4796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4950..5538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5030..5049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5709..6500,6501..6569))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6332..6351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    7162..7296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        7310..7689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggggggggg gggggggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga
       61 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
      121 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      181 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      241 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      301 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
      361 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg
      421 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg
      481 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc
      541 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg
      601 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc
      661 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac
      721 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg
      781 cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag
      841 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
      901 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
      961 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
     1021 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
     1081 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     1141 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
     1201 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattagc
     1261 caccatggat tacaaggatg acgacgataa gctcgagatg tctagactgg gagccctggg
     1321 tggtgcccgt gccgggctgg gactgttgct gggtaccgcc gccggccttg gattcctgtg
     1381 cctcctttac agccagcgat ggaaacggac ccagcgtcat ggccgcagcc agagcctgcc
     1441 caactccctg gactatacgc agacttcaga tcccggacgc cacgtgatgc tcctgcgggc
     1501 tgtcccaggt ggggctggag atgcctcagt gctgcccagc cttccacggg aaggacagga
     1561 gaaggtgctg gaccgcctgg actttgtgct gaccagcctt gtggcgctgc ggcgggaggt
     1621 ggaggagctg agaagcagcc tgcgagggct tgcgggggag attgttgggg aggtccgatg
     1681 ccacatggaa gagaaccaga gagtggctcg gcggcgaagg tttccgtttg tccgggagag
     1741 gagtgactac acaacagcca atgcggagtc tgacaatgag cgggactctg acaaagaaag
     1801 tgaggacggg gaagatgaag tgagctgtga gactgtgaag atggggagaa aggattctct
     1861 tgacttggag gaagaggcag cttcaggtgc ctccagtgcc ctggaggctg gaggttcctc
     1921 aggcttggag gatgtgctgc ccctcctgca gcaggccgac gagctgcaca ggggtgatga
     1981 gcaaggcaag cgggagggct tccagctgct gctcaacaac aagctggtgt atggaagccg
     2041 gcaggacttt ctctggcgcc tggcccgagc ctacagtgac atgtgtgagc tcactgagga
     2101 ggtgagcgag aagaagtcat atgccctaga tggaaaagaa gaagcagagg ctgctctgga
     2161 gaagggggat gagagtgctg actgtcacct gtggtatgcg gtgctttgtg gtcagctggc
     2221 tgagcatgag agcatccaga ggcgcatcca gagtggcttt agcttcaagg agcatgtgga
     2281 caaagccatt gctctccagc cagaaaaccc catggctcac tttcttcttg gcaggtggtg
     2341 ctatcaggtc tctcacctga gctggctaga aaaaaaaact gctacagcct tgcttgaaag
     2401 ccctctcagt gccactgtgg aagatgccct ccagagcttc ctaaaggctg aagaactaca
     2461 gccaggattt tccaaagcag gaagggtata tatttccaag tgctacagag aactagggaa
     2521 aaactctgaa gctagatggt ggatgaagtt ggccctggag ctgccagatg tcacgaagga
     2581 ggatttggct atccagaagg acctggaaga actggaagtc attttacgag actaggcggc
     2641 cgcgaattcc gccccccccc cctctccctc ccccccccct aacgttactg gccgaagccg
     2701 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     2761 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     2821 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     2881 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     2941 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     3001 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     3061 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     3121 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     3181 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     3241 caaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     3301 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     3361 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     3421 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     3481 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     3541 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     3601 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     3661 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     3721 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     3781 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     3841 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat
     3901 cgatgatcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3961 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4021 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4081 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     4141 gcggtgggct ctatggcttc tgaggcggaa agaacctgca gcccaagctt ggcgtaatca
     4201 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4261 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4321 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     4381 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4441 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4501 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4561 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4621 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4681 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     4741 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatgacc gctttggaaa
     6601 aacaaagact gtatttcctg gaaattaatg tttattcaat aaactgtgta ttcagctata
     6661 ttcacatagt ggtgaggctg aaatgaggcg ggaagaggcg gttggggctt aattatatca
     6721 atttgggtgg ccccacagcg cctccaaggc gccagtcctg ttttgacaag ttgcctctgg
     6781 aagcctctct tctttttctc cagagtaagc ggaggccagg ggcccccggc ctctgcttaa
     6841 tactaaaaaa aacagctgtt gtcatagtaa tgattgggtg gaaacattcc aggcctgggt
     6901 ggagaggctt tttgcttcct cttgcaaaac cacactgaca ttccaggcct gggtggagag
     6961 gctttttgct tcctcttgca aaaccacact gccctctgga gggcagttgc ctagcaacta
     7021 attaaaagag gatgtcgcac ggccagctgc ggtcagttag tcacttcctg cttaactgac
     7081 ttgacatttt ctattttaag agtcgggagg aaaattactg tgttggaggc cctccgccat
     7141 cttctgaagc tgaatcgaat taacttgttt attgcagctt ataatggtta caaataaagc
     7201 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     7261 tccaaactca tcaatgtatc ttatcatgtc tggatctgat atcatcgtcg acattgatta
     7321 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     7381 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     7441 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     7501 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     7561 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     7621 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     7681 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     7741 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     7801 gggggggggg gggg
//
