LOCUS       sequence-243544-        7351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human NPM-ALK fusion protein gene in pET28a bacterial expression 
            plasmid. Production of recombinant human NPM-ALK fusion protein in 
            E. coli Bl21 Rosetta2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7351)
  AUTHORS   Kourentzi K, Crum M, Patil U, Prebisch A, Chavan D, Vu B, Zeng Z, 
            Litvinov D, Zu Y, Willson RC
  TITLE     Recombinant expression, characterization, and quantification in 
            human cancer cell lines of the Anaplastic Large-Cell 
            Lymphoma-characteristic NPM-ALK fusion protein.
  JOURNAL   Sci Rep. 2020 Mar 19;10(1):5078. doi: 10.1038/s41598-020-61936-w.
  PUBMED    32193476
REFERENCE   2  (bases 1 to 7351)
  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: "Sci Rep."; 
            date: "2020-03-19"; volume: "10(1):5078. doi"; pages: " 
            10.1038/s41598-020-61936-w"
COMMENT     Sequence Label: pMCRW1
FEATURES             Location/Qualifiers
     source          1..7351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      62..517
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(149..168)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     359..380
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(609..1424)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(609..1424)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1332..1351
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1416..1435
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1546..2134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2035..2054
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2288..2305
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2320..2459
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2385..2404)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2545..2567
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2561..2752)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3561..4643)
                     /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     4605..4624
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4644..4721)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4927..4946
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5030..5049
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5030..5048
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5049..5073
                     /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)."
     RBS             5088..5110
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5129..5146
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5156..5173
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             7245..7262
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7315..7333)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      join(7329..7351,1..25)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
       61 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      121 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      181 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      241 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      301 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      361 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      421 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      481 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
      541 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      601 attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
      661 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
      721 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
      781 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
      841 gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca
      901 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
      961 cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca
     1021 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     1081 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     1141 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     1201 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     1261 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     1321 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     1381 aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     1441 ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc
     1501 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1561 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1621 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1681 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1741 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1801 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1861 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1921 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1981 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2041 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2101 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2161 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2221 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2281 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     2341 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
     2401 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     2461 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2521 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2581 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     2641 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     2701 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     2761 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     2821 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     2881 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     2941 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     3001 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     3061 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     3121 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     3181 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa
     3241 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     3301 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     3361 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     3421 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     3481 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     3541 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3601 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     3661 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     3721 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     3781 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     3841 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     3901 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     3961 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     4021 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     4081 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     4141 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     4201 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     4261 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     4321 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     4381 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     4441 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     4501 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     4561 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     4621 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     4681 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     4741 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     4801 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     4861 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     4921 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     4981 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     5041 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
     5101 aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca gcggcctggt
     5161 gccgcgcggc agccatatgg aagattcgat ggacatggac atgagccccc tgaggcccca
     5221 gaactatctt ttcggttgtg aactaaaggc cgacaaagat tatcacttta aggtggataa
     5281 tgatgaaaat gagcaccagt tatctttaag aacggtcagt ttaggggctg gtgcaaagga
     5341 tgagttgcac attgttgaag cagaggcaat gaattacgaa ggcagtccaa ttaaagtaac
     5401 actggcaact ttgaaaatgt ctgtacagcc aacggtttcc cttgggggct ttgaaataac
     5461 accaccagtg gtcttaaggt tgaagtgtgg ttcagggcca gtgcatatta gtggacagca
     5521 cttagtagtg taccgccgga agcaccagga gctgcaagcc atgcagatgg agctgcagag
     5581 ccctgagtac aagctgagca agctccgcac ctcgaccatc atgaccgact acaaccccaa
     5641 ctactgcttt gctggcaaga cctcctccat cagtgacctg aaggaggtgc cgcggaaaaa
     5701 catcaccctc attcggggtc tgggccatgg cgcctttggg gaggtgtatg aaggccaggt
     5761 gtccggaatg cccaacgacc caagccccct gcaagtggct gtgaagacgc tgcctgaagt
     5821 gtgctctgaa caggacgaac tggatttcct catggaagcc ctgatcatca gcaaattcaa
     5881 ccaccagaac attgttcgct gcattggggt gagcctgcaa tccctgcccc ggttcatcct
     5941 gctggagctc atggcggggg gagacctcaa gtccttcctc cgagagaccc gccctcgccc
     6001 gagccagccc tcctccctgg ccatgctgga ccttctgcac gtggctcggg acattgcctg
     6061 tggctgtcag tatttggagg aaaaccactt catccaccga gacattgctg ccagaaactg
     6121 cctcttgacc tgtccaggcc ctggaagagt ggccaagatt ggagacttcg ggatggcccg
     6181 agacatctac agggcgagct actatagaaa gggaggctgt gccatgctgc cagttaagtg
     6241 gatgccccca gaggccttca tggaaggaat attcacttct aaaacagaca catggtcctt
     6301 tggagtgctg ctatgggaaa tcttttctct tggatatatg ccatacccca gcaaaagcaa
     6361 ccaggaagtt ctggagtttg tcaccagtgg aggccggatg gacccaccca agaactgccc
     6421 tgggcctgta taccggataa tgactcagtg ctggcaacat cagcctgaag acaggcccaa
     6481 ctttgccatc attttggaga ggattgaata ctgcacccag gacccggatg taatcaacac
     6541 cgctttgccg atagaatatg gtccacttgt ggaagaggaa gagaaagtgc ctgtgaggcc
     6601 caaggaccct gagggggttc ctcctctcct ggtctctcaa caggcaaaac gggaggagga
     6661 gcgcagccca gctgccccac cacctctgcc taccacctcc tctggcaagg ctgcaaagaa
     6721 acccacagct gcagaggtct ctgttcgagt ccctagaggg ccggccgtgg aagggggaca
     6781 cgtgaatatg gcattctctc agtccaaccc tccttcggag ttgcacaagg tccacggatc
     6841 cagaaacaag cccaccagct tgtggaaccc aacgtacggc tcctggttta cagagaaacc
     6901 caccaaaaag aataatccta tagcaaagaa ggagccacac gacaggggta acctggggct
     6961 ggagggaagc tgtactgtcc cacctaacgt tgcaactggg agacttccgg gggcctcact
     7021 gctcctagag ccctcttcgc tgactgccaa tatgaaggag gtacctctgt tcaggctacg
     7081 tcacttccct tgtgggaatg tcaattacgg ctaccagcaa cagggcttgc ccttagaagc
     7141 cgctactgcc cctggagctg gtcattacga ggataccatt ctgaaaagca agaatagcat
     7201 gaaccagcct gggccctgat cccaagcttg cggccgcact cgagcaccac caccaccacc
     7261 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     7321 agcaataact agcataaccc cttggggcct c
//
