LOCUS       sequence-313580-        5728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect cell optimized TAK1-TAB1 construct used to solve structure 
            and deposit PDB ID: 4O91.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5728)
  AUTHORS   Tan L, Nomanbhoy T, Gurbani D, Patricelli M, Hunter J, Geng J, 
            Herhaus L, Zhang J, Pauls E, Ham Y, Choi HG, Xie T, Deng X, Buhrlage
            SJ, Sim T, Cohen P, Sapkota G, Westover KD, Gray NS
  TITLE     Discovery of type II inhibitors of TGFbeta-activated kinase 1 (TAK1)
            and mitogen-activated protein kinase kinase kinase kinase 2 
            (MAP4K2).
  JOURNAL   J Med Chem. 2015 Jan 8;58(1):183-96. doi: 10.1021/jm500480k. Epub 
            2014 Jul 30.
  PUBMED    25075558
REFERENCE   2  (bases 1 to 5728)
  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: "10.1021/jm500480k";
            journalName: "J Med Chem"; date: "2015-01-8- 8"; volume: "58"; 
            issue: "1"; pages: "183-96"
COMMENT     Sequence Label: LIC4B-His-TEV-TAK1-TAB1
FEATURES             Location/Qualifiers
     source          1..5728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        124..215
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     140..157
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             277..294
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             295..315
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    1456..1590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1493..1512)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1547..1566
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1619..1784
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1968..2422
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2055..2074)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2264..2285
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2449..2553
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2554..2622,2623..3414)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2772..2791)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3585..4173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4074..4093
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4327..4344
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4478..4702
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4769..5302)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5472..5477,5478..5494,5495..5500))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
       61 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggactattaa
      121 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
      181 ttcgtaacag ttttgtaata aaaaaaccta taactattcc ggattattca taccgtccca
      241 ccatcgggcg cggatcccgg tccgatgggt tcttctcacc atcaccatca ccatgaaaac
      301 ctgtacttcc aatccaatgc atcgttgcac atgatcgact acaaggaaat cgaggttgaa
      361 gaggtggtgg gaaggggcgc tttcggagtg gtctgtaagg ctaagtggcg tgctaaggac
      421 gttgccatca agcagatcga gtccgaatct gagcgcaagg ctttcatcgt ggaactgagg
      481 caactctcta gagtcaacca ccctaacatc gttaagctgt acggagcttg cctcaaccca
      541 gtgtgtttgg tcatggaata cgccgagggt ggcagcttgt acaacgtcct gcacggcgct
      601 gagccactgc cgtactacac cgctgctcac gctatgagct ggtgcctcca gtgttcacaa
      661 ggagttgctt acttgcactc catgcagccc aaggccctga tccaccgtga tctcaagccc
      721 cctaacctgc tcttggttgc tggaggtaca gtgttgaaga tctgcgactt cggcaccgcc
      781 tgtgatatcc aaacccacat gactaacaac aagggatcgg ctgcctggat ggctcctgaa
      841 gtgttcgagg gtagcaacta ctcagagaag tgcgacgtgt tctcctgggg tatcatcctc
      901 tgggaagtca tcacccgtcg caagccattc gatgagatcg gcggaccggc tttcaggatc
      961 atgtgggccg tgcacaacgg cactagacca ccgctcatca agaacttgcc caagcctatc
     1021 gagtcactga tgacacgttg ctggtcgaag gacccatccc agcgcccgtc tatggaggaa
     1081 atcgtcaaga tcatgactca cctcatgagg tacttccccg gtgccgacga acccttgcag
     1141 tacccttgtc aacacagcct gccccctggc gaggatggca gagttgagcc atacgttgat
     1201 ttcgccgagt tctacaggtt gtggagcgtt gaccacggag agtgataata acattggaag
     1261 tggataacgg atccgaattc gagcgccgtc gacaagcttg cggccgcact cgaggcatgc
     1321 ggtaccaagc ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag
     1381 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1441 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1501 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1561 ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag gagatccgaa
     1621 ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta
     1681 cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact
     1741 ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt
     1801 cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc
     1861 tactttttct ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat taatgtttgt
     1921 aattgactga atatcaacgc ttatttgcag cctgaatggc gaatgggacg cgccctgtag
     1981 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2041 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2101 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2161 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2221 gacggttttt cgccctttga cgttggagtc cacgttctta atagtggact cttgttccaa
     2281 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2341 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2401 aaactattaa cgtttacaat ttcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2461 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2521 taaatgcttc aatactattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2581 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2641 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2701 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2761 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2821 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2881 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2941 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3001 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3061 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3121 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3181 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3241 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3301 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3361 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3421 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3481 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3541 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3601 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3661 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3721 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3781 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3841 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3901 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3961 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4021 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4081 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4141 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4201 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4261 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4321 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     4381 acgcatctgt gcggtatttc acaccgcata gaccagccgc gtaacctggc aaaatcggtt
     4441 acggttgagt aataaatgga tgccctgcgt aagcgggtgt gggcggacaa taaagtctta
     4501 aactgaacaa aatagatcta aactatgaca ataaagtctt aaactagaca gaatagttgt
     4561 aaactgaaat cagtccagtt atgctgtgaa aaagcatact ggacttttgt tatggctaaa
     4621 gcaaactctt cattttctga agtgcaaatt gcccgtcgta ttaaagaggg gcgtggccaa
     4681 gggcatggta aagactatat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg
     4741 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     4801 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     4861 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     4921 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga
     4981 tatagatctc actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa
     5041 caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     5101 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     5161 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     5221 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     5281 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     5341 atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg
     5401 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct
     5461 acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt gccttcatcc
     5521 gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag gcatttctgt
     5581 cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca ttggcggcct
     5641 tgctgttctt ctacggcaag gtgctgtgca cggatctgcc ctggcttcag gagatcggaa
     5701 gacctcggcc gtcgcggcgc ttgccggt
//
