LOCUS       sequence-59331-e        8012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8012)
  AUTHORS   Besse A, Lamothe B, Campos AD, Webster WK, Maddineni U, Lin SC, Wu 
            H, Darnay BG
  TITLE     TAK1-dependent signaling requires functional interaction with 
            TAB2/TAB3.
  JOURNAL   J Biol Chem. 2007 Feb 9;282(6):3918-28. Epub 2006 Dec 8.
  PUBMED    17158449
REFERENCE   2  (bases 1 to 8012)
  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: "J Biol 
            Chem."; date: "2007-02-9"; volume: "282(6)"; pages: "3918-28. Epub 
            2006 Dec 8"
COMMENT     Sequence Label: p3XFLAG-TAK1-K63A
FEATURES             Location/Qualifiers
     source          1..8012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(1764..1775)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2695..2760
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2771..3393
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2970..2989)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3417..3437)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3435..3732
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3786..4580
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3840..3859)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4450..4469
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5232..5366
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5269..5288)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5323..5342
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5371..5390)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5372..5390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5404..5420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5404..5420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5417..5439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5428..5444
                     /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)."
     promoter        complement(join(5452..5458,5459..5476,5477..5482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5497..5518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5635..5652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5806..6394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5886..5905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6565..7356,7357..7425))
                     /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     7188..7207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7426..7530)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7557..7985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7644..7663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7854..7875
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaagcttg cggccgcgaa ttccaatgtc gacagcctcc
      961 gccgcctcgt cctcctcctc gtcttctgcc agtgagatga tcgaagcgcc gtcgcaggtc
     1021 ctgaacttcg aagagatcga ctacaaggag atcgaggtgg aagaggttgt cggaagagga
     1081 gcttttggag tagtttgcaa agctaagtgg agagcaaaag atgtcgctat tgcacagata
     1141 gaaagtgagt ctgagaggaa ggctttcatt gtggagctcc ggcagttgtc gcgtgtgaac
     1201 catcctaaca ttgtcaagtt gtacggagcc tgcctgaatc cagtatgtct tgtgatggaa
     1261 tatgcagagg ggggctcatt gtataatgtg ctgcatggtg ctgaaccatt gccttactac
     1321 actgctgctc atgccatgag ctggtgttta cagtgttccc aaggagtggc ttacctgcac
     1381 agcatgcagc ccaaagcgct gattcacagg gacctcaagc ctccaaactt gctgctggtt
     1441 gcaggaggga cagttctaaa aatctgcgat tttggtacag cttgtgacat ccaaacacac
     1501 atgaccaata ataaagggag tgctgcttgg atggcgcctg aagtgtttga aggtagcaat
     1561 tacagtgaaa agtgtgatgt cttcagctgg ggtattatcc tctgggaagt gataacacgc
     1621 cggaaaccct tcgatgagat cggtggccca gctttcagaa tcatgtgggc tgttcataat
     1681 ggcactcgac caccactgat caaaaattta cctaagccca ttgagagctt gatgacacgc
     1741 tgttggtcta aggacccatc tcagcgccct tcaatggagg aaattgtgaa aataatgact
     1801 cacttgatgc ggtacttccc aggagcggat gagccattac agtatccttg tcagtactct
     1861 gatgaagggc agagcaactc agccaccagc acaggctcgt tcatggacat tgcttctaca
     1921 aataccagta ataaaagtga cacaaatatg gaacaggttc ctgccacaaa cgacactatt
     1981 aaacgcttgg agtcaaaact gttgaaaaac caggcaaagc aacagagtga atctggacgc
     2041 ctgagcttgg gagcctctcg tgggagcagt gtggagagct tgccccccac ttccgagggc
     2101 aagaggatga gtgctgacat gtctgaaata gaagccagga tcgtggcgac tgcaggtaac
     2161 gggcaaccaa ggcgtagatc catccaagac ttgactgtta ctgggacaga acctggtcag
     2221 gtgagcagcc ggtcatccag ccctagtgtc agaatgatca ctacctcagg accaacctca
     2281 gagaagccag ctcgcagtca cccatggacc cctgatgatt ccacagacac caatggctca
     2341 gataactcca tcccaatggc gtatcttaca ctggatcacc agctacagcc tctagcgccg
     2401 tgcccaaact ccaaagaatc catggcagtg ttcgaacagc actgtaaaat ggcacaggag
     2461 tatatgaaag ttcaaaccga aatcgcattg ttactacaga gaaagcaaga actagttgca
     2521 gaattggacc aggatgaaaa ggaccagcaa aatacatctc gtctggtaca ggaacataaa
     2581 aagcttttag atgaaaacaa aagcctttct acttattacc agcaatgcaa aaaacaacta
     2641 gaggtcatca gaagccaaca gcagaaacga caaggcactt caggatcccg ggctgactac
     2701 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     2761 tagtgatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     2821 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     2881 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     2941 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     3001 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     3061 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     3121 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     3181 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     3241 tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac agcataggct
     3301 acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg
     3361 cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa
     3421 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3481 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3541 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3601 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3661 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3721 aggctttttt ggaggaattg atcagcttgg gatctgatca agagacagga tgaggatcgt
     3781 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3841 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3901 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3961 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4021 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4081 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4141 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4201 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4261 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4321 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4381 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4441 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4501 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4561 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4621 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4681 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4741 cttcgcccac cccgggctcg atcccctcgc gagttggttc agctgctgcc tgaggctgga
     4801 cgacctcgcg gagttctacc ggcagtgcaa atccgtcggc atccaggaaa ccagcagcgg
     4861 ctatccgcgc atccatgccc ccgaactgca ggagtgggga ggcacgatgg ccgctttggt
     4921 cgacccggac gggacgctcc tgcgcctgat acagaacgaa ttgcttgcag gcatctcatg
     4981 agtgtgtctt cccgttttcc gcctgaggtc actgcgtgga tggagcgctg gcgcctgctg
     5041 cgcgacggcg agctgctcac cacccactcg ccaagctgga accgtaaaaa ggccgcgttg
     5101 ctggcgtttt tccataggct ccgccgatca taatcagcca taccacattt gtagaggttt
     5161 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5221 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5281 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5341 tcaatgtatc ttatcatgtc tggatcaatt ccctatagtg agtcgtatta aattcgtaat
     5401 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5461 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5521 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5581 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5641 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5701 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5761 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5821 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5881 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5941 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6001 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6061 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6121 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6181 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6241 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6301 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6361 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6421 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6481 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6541 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6601 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6661 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6721 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6781 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6841 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6901 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6961 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7021 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7081 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7141 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7201 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7261 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7321 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7381 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7441 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7501 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     7561 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     7621 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     7681 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     7741 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     7801 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     7861 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     7921 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     7981 gaattttaac aaaatattaa cgcttacaat tt
//
