LOCUS       sequence-217680-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For protein expression and purification of Drosophila pumilio (amino
            acids 566-1090).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6558)
  AUTHORS   Hara M, Lourido S, Petrova B, Lou HJ, Von Stetina JR, Kashevsky H, 
            Turk BE, Orr-Weaver TL
  TITLE     Identification of PNG kinase substrates uncovers interactions with 
            the translational repressor TRAL in the oocyte-to-embryo transition.
  JOURNAL   Elife. 2018 Feb 26;7. pii: 33150. doi: 10.7554/eLife.33150.
  PUBMED    29480805
REFERENCE   2  (bases 1 to 6558)
  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: "Elife."; 
            date: "2018-02-26"; volume: "7. pii: 33150. doi"; pages: " 
            10.7554/eLife.33150"
COMMENT     Sequence Label: pGEX6P-1 PUM FL
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        833..910
                     /gene="lacI"
                     /label=lacI promoter
     CDS             911..1993
                     /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     complement(930..949)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2042..2047,2048..2065,2066..2072)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2080..2096
                     /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)."
     primer_bind     2104..2120
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2104..2120
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2116..2643
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2136..2153)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2136..2152)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2145..2167)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2745..2750,2751..2766,2767..2773)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2781..2797
                     /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)."
     CDS             2820..3473
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3431..3453
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3480..3503
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(5170..5192)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5337..5355)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5423..5527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5528..5596,5597..6388)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5746..5765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat
      841 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
      901 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
      961 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     1021 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     1081 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     1141 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     1201 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     1261 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     1321 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     1381 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     1441 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     1501 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     1561 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     1621 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     1681 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     1741 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     1801 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     1861 gctgttgccc gtctcactgg tgaaaagaaa aaccacgcta gcgcccaata cgcaaaccgc
     1921 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     1981 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg
     2041 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     2101 acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt
     2161 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2221 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2281 aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg
     2341 gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt
     2401 tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt
     2461 cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta
     2521 caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg
     2581 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     2641 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     2701 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
     2761 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagtattca
     2821 tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
     2881 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
     2941 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
     3001 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
     3061 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
     3121 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
     3181 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
     3241 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
     3301 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
     3361 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
     3421 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc
     3481 tggaagttct gttccagggg cccctgggat ccccggaatt catgctcatg gatcatggcc
     3541 aaggcatggg aggcggcttg ggcggagttg ttggatctgg caacggagtc ggcggtggca
     3601 gcggcggagg cggggcaggc ggcgcttatg cggcccacca gcagatggcc gcccagatga
     3661 gtcaattgca accgccgatg atgaacggcg ttggcggcgg aatgccaatg gcagcacagt
     3721 caccaatgtt gaatcaccag gcagctggac ccaatcatat ggaatctccc ggaaatctct
     3781 tgcagcagca aaattttgat gttcagcaac tgtttcgctc gcagaatccg ggcctagcag
     3841 cagttgccac aaatgcagcg gccgcagcag cagccgcagc agctgccaca tcggcagcga
     3901 gtgctgcggc agcggtgggc gcaccacccg ttcccaacgg atcgctgcag cagtcgcagc
     3961 agcaacagca gcagcagcaa caacagcagc agcaacaaca gatgcacatg gcggccgcgt
     4021 cgcaacaatt tttggccgcc cagcagcagg cgcaaaatgc ggcctatgcc gcccaacagg
     4081 ccacgtccta cgtcatcaat ccgggccagg aggctgcccc gtatatgggc atgattgccg
     4141 ccgcccagat gccgtactat ggcgtagcac catggggcat gtatccgggc aatctgattc
     4201 cgcaacaggg aacgcagccg cgccgccccc tcaccccctc gcagcagggt gccgagaatc
     4261 agccgtatca ggtcatcccg gcattcctcg atcacacggg ctccttgctg atgggaggac
     4321 ctcgcaccgg gacgccgatg cgtctggtta gccccgcccc cgttctggtg cccccgggcg
     4381 ctacccgtgc cggccccccg cccccgcagg ggccacagct gtatcagccg cagccgcaga
     4441 cggcccaaca gaatctctac tcgcagcaga atggatccag tgtcggaggc ctcgccttga
     4501 acacgagctc gttgacgggt cgccgcgact ccttcgaccg cagcacctcc gccttcagtc
     4561 cctcgaccat ggactacacc agcagcggtg tggcagcggc cgccaatgcg gtgaacagca
     4621 cagtggccca ggcagcagca gctgccgcag cagccgccgc agcgcgtggc aagtggccgg
     4681 gagcgatgtc gggagcggcc agtggagcct acggagccct gggagcgggc aatgcttcgg
     4741 ccagtcccct gggcgcacca atcacgccgc cgccatcggc gcaatcctgt ctcctgggca
     4801 gtcgggcacc tggagccgag tcccgccagc ggcagcagca acaacagcag ctggccgccg
     4861 ttggtctgcc ggcgactgca gcagctgctc aggcagcggt ggccgcggct gccaacaata
     4921 tgttcggatc caacagctcg atcttctcga atcccctggc cattccgggt accgcagctg
     4981 tggcagctgc agcggcagca gcagcggccg ccaactcgcg tcaggtggct gccacggcag
     5041 cggcagcagc ggcggtggca gcagcagccg gcggagtggg aggtgcccca cagccataag
     5101 tcgactcgag cggccgcatc gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga
     5161 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5221 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     5281 agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac gaaagggcct
     5341 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5401 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5461 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5521 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5581 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5641 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5701 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5761 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5821 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5881 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5941 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6001 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6061 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6121 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6181 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6241 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6301 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6361 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6421 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6481 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6541 aaagatcaaa ggatcttc
//
