LOCUS       sequence-236089-        6939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid containing His and MBP tags for 3 PRM 
            motif repeats of Human Abl..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  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 6939)
  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 3R
FEATURES             Location/Qualifiers
     source          1..6939
                     /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             1522..1542
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1543..1560
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1573..1590)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1573..1589)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1582..1604)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1768..1785)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1938..2024
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2116..2143
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2162..2253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2254..2322,2323..3114)
                     /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(2472..2491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3156..3669)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3351..3372)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3563..3582
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3780..4368
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4269..4288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4522..4539
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4645..4664)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4805..4827
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4821..5012)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        5508..5585
                     /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             5586..6668
                     /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(5605..5624)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6911..6916,6917..6932,6933..6939)
                     /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 gctcaggatc cgaaaacctg tattttcagg gccatcatca tcatcatcac
     1561 tgaaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     1621 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     1681 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc agcttggctg
     1741 ttttggcgga tgagataaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     1801 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     1861 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
     1921 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1981 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     2041 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     2101 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     2161 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2221 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2281 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2341 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2401 aacagcggta agatccttga gagttttcgc cccgaagaac gtttcccaat gatgagcact
     2461 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     2521 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2581 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2641 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2701 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2761 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2821 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2881 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2941 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3001 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3061 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3121 gaccaagttt actcatatat actttagatt gatttacccc ggttgataat cagaaaagcc
     3181 ccaaaaacag gaagattgta taagcaaata tttaaattgt aaacgttaat attttgttaa
     3241 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3301 aaatccctta taaatcaaaa gaatagcccg agatagggtt gagtgttgtt ccagtttgga
     3361 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3421 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
     3481 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3541 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3601 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3661 agggcgcgta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3721 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3781 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3841 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3901 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3961 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4021 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4081 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4141 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4201 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4261 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4321 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4381 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4441 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4501 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4561 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatgtggtg cactctcagt
     4621 acaccgcatg tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4681 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4741 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4801 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4861 ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac agatgtctgc ctgttcatcc
     4921 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     4981 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     5041 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     5101 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     5161 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     5221 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     5281 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     5341 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     5401 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     5461 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg aaattccgac accatcgaat
     5521 ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     5581 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     5641 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     5701 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     5761 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     5821 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     5881 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     5941 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     6001 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     6061 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     6121 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     6181 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     6241 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     6301 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     6361 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     6421 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     6481 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     6541 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     6601 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6661 ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcaca attctcatgt
     6721 ttgacagctt atcatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg
     6781 aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc
     6841 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg
     6901 aaatgagctg ttgacaatta atcatcggct cgtataatg
//
