LOCUS       sequence-217568-        7513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For bacterial expression of MBP fusion of C terminal region of 
            Drosophila Tral.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7513)
  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 7513)
  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: pMAL c2x Tral C
FEATURES             Location/Qualifiers
     source          1..7513
                     /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"
     misc_feature    775..917
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(843..862)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1003..1025
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1019..1210)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        1706..1783
                     /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."
     CDS             1784..2866
                     /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(1803..1822)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3109..3114,3115..3130,3131..3137)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3145..3161
                     /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     3150..3172
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             3231..4331
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     4305..4328
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4380..4391
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5307..5324)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5307..5323)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5316..5338)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5502..5519)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5672..5758
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5850..5877
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5896..5987
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5988..6056,6057..6848)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6206..6225)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6890..7403)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7085..7106)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7297..7316
                     /label=F1ori-R
                     /note="F1 origin, 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 cgcatatatg gtgcactctc
      841 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
      901 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
      961 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     1021 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     1081 ggtcgtgcag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     1141 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     1201 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     1261 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     1321 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     1381 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     1441 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     1501 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     1561 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     1621 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc
     1681 aggacccaac gctgcccgaa attccgacac catcgaatgg tgcaaaacct ttcgcggtat
     1741 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     1801 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     1861 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     1921 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     1981 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     2041 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     2101 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     2161 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     2221 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     2281 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     2341 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     2401 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     2461 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     2521 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     2581 ggatatctca gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     2641 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     2701 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     2761 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2821 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2881 atgtaagtta gctcactcat taggcacaat tctcatgttt gacagcttat catcgactgc
     2941 acggtgcacc aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag
     3001 gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt
     3061 tttgcgccga catcataacg gttctggcaa atattctgaa atgagctgtt gacaattaat
     3121 catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca ggaaacagcc
     3181 agtccgttta ggtgttttca cgagcacttc accaacaagg accatagcat atgaaaatcg
     3241 aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt ctcgctgaag
     3301 tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat ccggataaac
     3361 tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt atcttctggg
     3421 cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc accccggaca
     3481 aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac aacggcaagc
     3541 tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa gatctgctgc
     3601 cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg aaagcgaaag
     3661 gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg ctgattgctg
     3721 ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa gacgtgggcg
     3781 tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt aaaaacaaac
     3841 acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa ggcgaaacag
     3901 cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa gtgaattatg
     3961 gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga
     4021 gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc ctcgaaaact
     4081 atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag
     4141 cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc actatggaaa
     4201 acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc tggtatgccg
     4261 tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag
     4321 acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac aacctcggga
     4381 tcgagggaag gatttcagaa ttcatgcatc gcggcaacca gaactcgaac aacatgatga
     4441 acatgcgcaa ccgcggcatg ggatcccgcg gccccatgcg acccaatcag ggctatcgtc
     4501 cgcagtcggc caataatcag aacaagccgc ggaacaagat caagttcgag ggcgacttcg
     4561 atttcgagca ggcaaacaac aagttcgagg aactgcgctc ccaactggcc aagctcaagg
     4621 tggccgagga tggtgcaccc aagccagcca ccaatgcaac ggccgccact gcaactgcaa
     4681 ccaatgagca ggtgggtgag aaggttgaag gcgttcacac actgaatggc gagaccgaca
     4741 agaaggatga ttctggcaac gagaccggcg ctggagagca cgagcctgag gaggatgatg
     4801 ttgctgtgtg ctacgacaag accaaatcgt tcttcgacaa catctcgtgc gaggctgccc
     4861 aggatcgcag caagaacaag aagaacgatt ggcgccagga gcgcaagttg aacacggaga
     4921 ccttcggagt gtcctccaca cgacgtggca gttaccgcgg acgcaaccaa tactacaaca
     4981 ataatggcaa cggcggcatt aactcgggat atggaggagc gccaggctac aacaggaaca
     5041 attatcgcat gggtggcggc ggcggaaact tccgaaacag gagcaacaat cgcaacaacg
     5101 gcggcggtcg tggcggaaac ggaatgccaa acatcaccaa tggcaacacg gctgctgcgc
     5161 tgaaggcggc caacaatgct gctggccacg gatccaatgc cacggactcc agtgcaccaa
     5221 atgccacaac cgcgacgaca aagtcgacgt ccctcttgcc agagcagacg caacaggtgg
     5281 cggcagtttc acaatagaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa
     5341 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     5401 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5461 tggcagcttg gctgttttgg cggatgagat aagattttca gcctgataca gattaaatca
     5521 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     5581 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     5641 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     5701 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     5761 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     5821 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     5881 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5941 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6001 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6061 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6121 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgtttcc
     6181 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     6241 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6301 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6361 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6421 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6481 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6541 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6601 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6661 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6721 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6781 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6841 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta ccccggttga
     6901 taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa ttgtaaacgt
     6961 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     7021 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     7081 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     7141 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt
     7201 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     7261 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     7321 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     7381 taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa gatccttttt gataatctca
     7441 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7501 tcaaaggatc ttc
//
