LOCUS       sequence-236084-        7014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid containing His and MBP tags for 4 PRM 
            motif repeats of Human Abl..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7014)
  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 7014)
  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 4R
FEATURES             Location/Qualifiers
     source          1..7014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..24
                     /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     13..35
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             94..1194
                     /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      1051..1060
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1168..1191
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1243..1254
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1267..1287
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1597..1617
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1618..1635
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1648..1665)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1648..1664)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1657..1679)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1843..1860)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2013..2099
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2191..2218
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2237..2328
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2329..2397,2398..3189)
                     /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(2547..2566)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3231..3744)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3426..3447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3638..3657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3855..4443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4344..4363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4597..4614
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4720..4739)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4880..4902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4896..5087)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        5583..5660
                     /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             5661..6743
                     /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(5680..5699)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6986..6991,6992..7007,7008..7014)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 tgtggaattg tgagcggata acaatttcac acaggaaaca gccagtccgt ttaggtgttt
       61 tcacgagcac ttcaccaaca aggaccatag attatgaaaa ctgaagaagg taaactggta
      121 atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag
      181 aaagataccg gaattaaagt caccgttgag catccggata aactggaaga gaaattccca
      241 caggttgcgg caactggcga tggccctgac attatcttct gggcacacga ccgctttggt
      301 ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag
      361 ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc
      421 gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc
      481 tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg
      541 ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg
      601 ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg
      661 aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc
      721 gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc
      781 ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg
      841 accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc
      901 gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa
      961 ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag
     1021 gaagagttgg cgaaagatcc acgtattgcc gccaccatgg aaaacgccca gaaaggtgaa
     1081 atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc
     1141 aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg
     1201 agctcgaaca acaacaacaa taacaataac aacaacctcg ggatcgaggg aaggatttca
     1261 gaatttgaaa acctgtattt tcagggccat atgaaaggag gcagctgggg gggtagcaag
     1321 aaaaagaaaa ccgcaccaac cccgcctaaa cgtagcggcg gctcaggtgg ttctgggggt
     1381 agcggaggaa gtaagaaaaa gaaaaccgca ccaaccccgc ctaaacgtag cggcggctca
     1441 ggtggttctg ggggtagcgg aggaagtaag aaaaagaaaa ccgcaccaac cccgcctaaa
     1501 cgtagcggcg gctcaggtgg ttctgggggt agcggaggaa gtaagaaaaa gaaaaccgca
     1561 ccaaccccgc ctaaacgtag cggcggctca ggatccgaaa acctgtattt tcagggccat
     1621 catcatcatc atcactgaaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga
     1681 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1741 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1801 atggcagctt ggctgttttg gcggatgaga taagattttc agcctgatac agattaaatc
     1861 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
     1921 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
     1981 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     2041 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     2101 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
     2161 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
     2221 cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2281 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2341 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2401 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2461 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttc
     2521 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     2581 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2641 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2701 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2761 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2821 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2881 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2941 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3001 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3061 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3121 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3181 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt accccggttg
     3241 ataatcagaa aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaaacg
     3301 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3361 aggccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata gggttgagtg
     3421 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3481 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt
     3541 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3601 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3661 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3721 ttaatgcgcc gctacagggc gcgtaaaagg atctaggtga agatcctttt tgataatctc
     3781 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3841 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3901 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3961 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4021 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4081 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4141 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4201 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4261 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4321 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4381 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4441 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4501 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4561 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4621 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcatg
     4681 tggtgcactc tcagtacacc gcatgtggtg cactctcagt acaatctgct ctgatgccgc
     4741 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     4801 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4861 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4921 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgcagcga ttcacagatg
     4981 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     5041 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     5101 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     5161 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     5221 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     5281 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     5341 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     5401 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     5461 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     5521 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgaaatt
     5581 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     5641 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     5701 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     5761 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     5821 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     5881 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     5941 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     6001 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     6061 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     6121 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     6181 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     6241 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     6301 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     6361 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     6421 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     6481 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     6541 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     6601 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     6661 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6721 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     6781 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     6841 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     6901 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     6961 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatg
//
