LOCUS       sequence-338856-        5483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cleavable His-tagged PTPIP51_Delta-TPR domain (36-235) in 
            E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5483)
  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 5483)
  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: pQlinkH-PTPIP51(36-235)
FEATURES             Location/Qualifiers
     source          1..5483
                     /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             147..167
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    180..204
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             204..224
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(844..868)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      890..984
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1078..1737
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1144..1163)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1802..1888
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1981..3063)
                     /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     3025..3044
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3064..3141)
                     /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(3225..3247)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3333..3473
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3388..3407
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3488..3505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3659..4247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3739..3758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4418..5209,5210..5278))
                     /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     5041..5060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5279..5383)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5451..5469
                     /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 atgaaacatc accatcacca tcaccatggt gctgcaggta
      181 caagtttgta caaaaaagca ggcgagaatc tttattttca gggatccaaa cggacccagc
      241 gtcatggccg cagccagagc ctgcccaact ccctggacta tacgcagact tcagatcccg
      301 gacgccacgt gatgctcctg cgggctgtcc caggtggggc tggagatgcc tcagtgctgc
      361 ccagccttcc acgggaagga caggagaagg tgctggaccg cctggacttt gtgctgacca
      421 gccttgtggc gctgcggcgg gaggtggagg tgctgagaag cagcctgcga gggcttgcgg
      481 gggagattgt tggggaggtc cgatgccaca tggaagagaa ccagagagtg gctcggcggc
      541 gaaggtttcc gtttgtccgg gagaggagtg actccactgg ctccagctct gtctacttca
      601 cggcctcctc gggagccacg ttcacagatg ctgagagtga agggggttac acaacagcca
      661 atgcggagtc tgacaatgag cgggactctg acaaagaaag tgaggacggg gaagatgaag
      721 tgagctgtga gactgtgaag atggggagaa aggattctct tgacttggag gaagaggcag
      781 cttcaggtgc ctccagtgcc ctggaggctg gaggttcctc aggcttgtag gcggccgcct
      841 aggacccagc tttcttgtac aaagtggtaa gcttaattag ctgagcttgg actcctgttg
      901 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
      961 cgggcgtttt ttattggtga gaatccaagt aacaacacca tttaaatgga gtggttacaa
     1021 atggagtggt taattaaggc tagcttggcg agattttcag gagctaagga agctaaaatg
     1081 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     1141 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     1201 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     1261 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
     1321 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     1381 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     1441 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     1501 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     1561 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     1621 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     1681 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     1741 ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca
     1801 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1861 ggtgaacgct ctcctgagta ggacaaatcc gccctctaga ttacgtgcag tcgatgataa
     1921 gctgtcaaac atgagaattg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     1981 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2041 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     2101 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     2161 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     2221 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     2281 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     2341 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     2401 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     2461 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     2521 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     2581 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     2641 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     2701 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     2761 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     2821 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     2881 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     2941 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     3001 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     3061 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     3121 tttgcaccat tcgatggtgt cggaatttcg ggcagcgttg ggtcctggcc acgggtgcgc
     3181 atgatctaga gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     3241 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     3301 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     3361 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     3421 acatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     3481 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3541 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3601 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3661 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3721 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3781 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3841 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3901 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3961 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4021 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4081 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4141 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4201 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4261 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4321 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4381 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4441 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4501 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4561 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4621 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4681 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4741 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4801 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4861 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4921 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4981 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5041 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5101 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5161 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5221 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5281 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5341 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5401 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5461 tcacgaggcc ctttcgtctt cac
//
