LOCUS       sequence-236080-        7089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid containing His and MBP tags for 5 PRM 
            motif repeats of Human Abl..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7089)
  AUTHORS   Li P, Banjade S, Cheng HC, Kim S, Chen B, Guo L, Llaguno M, 
            Hollingsworth JV, King DS, Banani SF, Russo PS, Jiang QX, Nixon BT, 
            Rosen MK
  TITLE     Phase transitions in the assembly of multivalent signalling 
            proteins.
  JOURNAL   Nature. 2012 Mar 7;483(7389):336-40. doi: 10.1038/nature10879.
  PUBMED    22398450
REFERENCE   2  (bases 1 to 7089)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2012-03-7- 7"; volume: "483"; issue: "7389"; pages:
            "336-40"
COMMENT     Sequence Label: pMal-Abl-PRM 5R
FEATURES             Location/Qualifiers
     source          1..7089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             123..1223
                     /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="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1080..1089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1197..1220
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1272..1283
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1296..1316
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1701..1721
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1722..1739
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1752..1769)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1752..1768)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1761..1783)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1947..1964)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2117..2203
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2295..2322
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2341..2432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2433..2501,2502..3293)
                     /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(2651..2670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3335..3848)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3530..3551)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3742..3761
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3959..4547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4448..4467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4701..4718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4824..4843)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4984..5006
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5000..5191)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        5687..5764
                     /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             5765..6847
                     /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(5784..5803)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaac tgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg
      181 gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc
      241 atccggataa actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca
      301 ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa
      361 tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt
      421 acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca
      481 aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac
      541 tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc
      601 cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta
      661 aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga
      721 ttaaaaacaa acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata
      781 aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca
      841 aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg
      901 ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt
      961 tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc
     1021 tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg
     1081 ccaccatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt
     1141 tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg
     1201 aagccctgaa agacgcgcag actaattcga gctcgaacaa caacaacaat aacaataaca
     1261 acaacctcgg gatcgaggga aggatttcag aatttgaaaa cctgtatttt cagggccata
     1321 tgaaaggagg cagctggggg ggtagcaaga aaaagaaaac cgcaccaacc ccgcctaaac
     1381 gtagcggcgg ctcaggtggt tctgggggta gcggaggaag taagaaaaag aaaaccgcac
     1441 caaccccgcc taaacgtagc ggcggctcag gtggttctgg gggtagcgga ggaagtaaga
     1501 aaaagaaaac cgcaccaacc ccgcctaaac gtagcggcgg ctcaggtggt tctgggggta
     1561 gcggaggaag taagaaaaag aaaaccgcac caaccccgcc taaacgtagc ggcggctcag
     1621 gtggttctgg gggtagcgga ggaagtaaga aaaagaaaac cgcaccaacc ccgcctaaac
     1681 gtagcggcgg ctcaggatcc gaaaacctgt attttcaggg ccatcatcat catcatcact
     1741 gaaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     1801 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     1861 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggca gcttggctgt
     1921 tttggcggat gagataagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1981 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     2041 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     2101 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2161 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     2221 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     2281 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     2341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2401 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2461 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2521 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2581 acagcggtaa gatccttgag agttttcgcc ccgaagaacg tttcccaatg atgagcactt
     2641 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2701 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2761 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2821 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2881 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2941 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3001 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3061 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3121 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3181 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3241 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3301 accaagttta ctcatatata ctttagattg atttaccccg gttgataatc agaaaagccc
     3361 caaaaacagg aagattgtat aagcaaatat ttaaattgta aacgttaata ttttgttaaa
     3421 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3481 aatcccttat aaatcaaaag aatagcccga gatagggttg agtgttgttc cagtttggaa
     3541 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3601 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg
     3661 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3721 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3781 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3841 gggcgcgtaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3901 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3961 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4021 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4081 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4141 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4201 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4261 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4321 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4381 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4441 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4501 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4561 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4621 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4681 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     4741 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catgtggtgc actctcagta
     4801 caccgcatgt ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     4861 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     4921 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4981 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     5041 tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg
     5101 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     5161 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     5221 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     5281 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     5341 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     5401 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     5461 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     5521 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     5581 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     5641 cgatcatgcg cacccgtggc caggacccaa cgctgcccga aattccgaca ccatcgaatg
     5701 gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     5761 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     5821 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     5881 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     5941 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     6001 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     6061 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     6121 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     6181 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     6241 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     6301 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     6361 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     6421 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     6481 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     6541 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
     6601 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     6661 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     6721 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     6781 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     6841 gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcacaa ttctcatgtt
     6901 tgacagctta tcatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga
     6961 agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca
     7021 ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga
     7081 aatgagctg
//
