LOCUS       sequence-338871-        6224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cleavable GST-tagged PTPIP51 TPR domain (236-470) in E. 
            Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  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 6224)
  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: pQlinkG2-PTPIP51(236-470)
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(36..50,51..56,57..73,74..80)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    56..72
                     /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)."
     protein_bind    88..104
                     /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)."
     RBS             123..134
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             141..794
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     752..774
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    816..840
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             840..860
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1585..1609)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1631..1725
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1819..2478
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1885..1904)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2543..2629
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2722..3804)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     3766..3785
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3805..3882)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3966..3988)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4074..4214
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4129..4148
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4229..4246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4400..4988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4480..4499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5159..5950,5951..6019))
                     /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     5782..5801
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6020..6124)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6192..6210
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagctta attaacaaca ccatttgtcg agaaatcata aaaaatttat ttgctttgtg
       61 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca
      121 ttaaagagga gaaattaact atgtccccta tactaggtta ttggaaaatt aagggccttg
      181 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg
      241 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca
      301 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt
      361 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa
      421 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag
      481 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg
      541 aagatcgttt atgtcataaa acatatttga atggtgatca tgtaacccat cctgacttca
      601 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc
      661 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga
      721 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg
      781 accatcctcc aaaatcggat ctgggtgctg caggtacaag tttgtacaaa aaagcaggcg
      841 agaatcttta ttttcaggga tccgaggatg tgctgcccct cctgcagcag gccgacgagc
      901 tgcacagggg tgatgagcaa ggcaagcggg agggcttcca gctgctgctc aacaacaagc
      961 tggtgtatgg aagccggcag gactttctct ggcgcctggc ccgagcctac agtgacatgt
     1021 gtgagctcac tgaggaggtg agcgagaaga agtcatatgc cctagatgga aaagaagaag
     1081 cagaggctgc tctggagaag ggggatgaga gtgctgactg tcacctgtgg tatgcggtgc
     1141 tttgtggtca gctggctgag catgagagca tccagaggcg catccagagt ggctttagct
     1201 tcaaggagca tgtggacaaa gccattgctc tccagccaga aaaccccatg gctcactttc
     1261 ttcttggcag gtggtgctat caggtctctc acctgagctg gctagaaaaa aaaactgcta
     1321 cagccttgct tgaaagccct ctcagtgcca ctgtggaaga tgccctccag agcttcctaa
     1381 aggctgaaga actacagcca ggattttcca aagcaggaag ggtatatatt tccaagtgct
     1441 acagagaact agggaaaaac tctgaagcta gatggtggat gaagttggcc ctggagctgc
     1501 cagatgtcac gaaggaggat ttggctatcc agaaggacct ggaagaactg gaagtcattt
     1561 tacgagacta ggcggccgcc taggacccag ctttcttgta caaagtggta agcttaatta
     1621 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     1681 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag taacaacacc
     1741 atttaaatgg agtggttaca aatggagtgg ttaattaagg ctagcttggc gagattttca
     1801 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     1861 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     1921 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     1981 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt
     2041 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     2101 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     2161 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     2221 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     2281 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     2341 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     2401 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     2461 tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggggta
     2521 atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     2581 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccctctag
     2641 attacgtgca gtcgatgata agctgtcaaa catgagaatt gtgcctaatg agtgagctaa
     2701 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2761 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     2821 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     2881 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     2941 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     3001 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     3061 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     3121 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     3181 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     3241 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     3301 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     3361 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     3421 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     3481 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     3541 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     3601 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     3661 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     3721 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     3781 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     3841 tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tcggaatttc gggcagcgtt
     3901 gggtcctggc cacgggtgcg catgatctag agctgcctcg cgcgtttcgg tgatgacggt
     3961 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     4021 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     4081 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     4141 agattgtact gagagtgcac cacatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4201 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4261 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4321 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4381 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4441 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4501 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4561 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4621 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4681 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4741 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4801 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4861 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4921 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4981 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5041 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5101 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5161 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5221 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5281 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5341 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5401 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5461 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5521 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5581 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5641 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5701 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5761 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5821 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5881 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5941 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6001 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6061 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6121 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     6181 taacctataa aaataggcgt atcacgaggc cctttcgtct tcac
//
