LOCUS       sequence-424654-        7131 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 7131)
  AUTHORS   Blonska M, Shambharkar PB, Kobayashi M, Zhang D, Sakurai H, Su B, 
            Lin X
  TITLE     TAK1 is recruited to the tumor necrosis factor-alpha (TNF-alpha) 
            receptor 1 complex in a receptor-interacting protein (RIP)-dependent
            manner and cooperates with MEKK3 leading to NF-kappaB activation.
  JOURNAL   J Biol Chem. 2005 Dec 30;280(52):43056-63. Epub 2005 Oct 31.
  PUBMED    16260783
REFERENCE   2  (bases 1 to 7131)
  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: "2005-12-30"; volume: "280(52)"; pages: "43056-63. 
            Epub 2005 Oct 31"
COMMENT     Sequence Label: pcDNA3 TAK1/F
FEATURES             Location/Qualifiers
     source          1..7131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      96..231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     158..177
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             312..1106
                     /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(366..385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     976..995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1280..1401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1317..1336)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1371..1390
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1450..1466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1450..1466)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1463..1485)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1474..1490
                     /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(1498..1504,1505..1522,1523..1528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1543..1564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1681..1698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1852..2440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1932..1951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2611..3402,3403..3471))
                     /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     3234..3253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3472..3576)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3651..3670)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3842..4221
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4222..4425
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4376..4396
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4470..4489
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4470..4488
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(5346..5357)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6277..6300
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(6329..6346)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6335..6559
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6605..7033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6692..6711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6902..6923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7042..7062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(7047..7131,1..245)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      121 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      181 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      241 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
      301 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      361 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      421 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      481 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      541 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      601 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      661 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
      721 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
      781 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
      841 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
      901 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
      961 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1021 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1081 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1141 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     1201 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     1261 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     1321 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1381 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     1441 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1501 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1561 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1621 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1681 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1741 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1801 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1981 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2041 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2101 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2161 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2281 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2521 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2581 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2641 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2701 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2761 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2821 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2881 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2941 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3001 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3061 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3601 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     3661 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     3721 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     3781 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     3841 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     3901 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     3961 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     4021 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     4081 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     4141 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     4201 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     4261 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     4321 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     4381 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga
     4441 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct
     4501 ggctagcgtt taaacgggcc ctctagaatg tctacagcct ctgccgcctc gtcctcctcc
     4561 tcgtcttctg ccagtgagat gatcgaagcg ccgtcgcagg tcctgaactt cgaagagatc
     4621 gactacaagg agatcgaggt ggaagaggtt gtcggaagag gagcttttgg agtagtttgc
     4681 aaagctaagt ggagagcaaa agatgtcgct attaaacaga tagaaagtga gtctgagagg
     4741 aaggctttca ttgtggagct ccggcagttg tcgcgtgtga accatcctaa cattgtcaag
     4801 ttgtacggag cctgcctgaa tccagtatgt cttgtgatgg aatatgcaga ggggggctca
     4861 ttgtataatg tgctgcatgg tgctgaacca ttgccttact acactgctgc tcatgccatg
     4921 agctggtgtt tacagtgttc ccaaggagtg gcttacctgc acagcatgca gcccaaagcg
     4981 ctgattcaca gggacctcaa gcctccaaac ttgctgctgg ttgcaggagg gacagttcta
     5041 aaaatctgcg attttggtac agcttgtgac atccaaacac acatgaccaa taataaaggg
     5101 agtgctgctt ggatggcgcc tgaagtgttt gaaggtagca attacagtga aaagtgtgat
     5161 gtcttcagct ggggtattat cctctgggaa gtgataacac gccggaaacc cttcgatgag
     5221 atcggtggcc cagctttcag aatcatgtgg gctgttcata atggcactcg accaccactg
     5281 atcaaaaatt tacctaagcc cattgagagc ttgatgacac gctgttggtc taaggaccca
     5341 tctcagcgcc cttcaatgga ggaaattgtg aaaataatga ctcacttgat gcggtacttc
     5401 ccaggagcgg atgagccatt acagtatcct tgtcagtact ctgatgaagg gcagagcaac
     5461 tcagccacca gcacaggctc gttcatggac attgcttcta caaataccag taataaaagt
     5521 gacacaaata tggaacaggt tcctgccaca aacgacacta ttaaacgctt ggagtcaaaa
     5581 ctgttgaaaa accaggcaaa gcaacagagt gaatctggac gcctgagctt gggagcctct
     5641 cgtgggagca gtgtggagag cttgcccccc acttccgagg gcaagaggat gagtgctgac
     5701 atgtctgaaa tagaagccag gatcgtggcg actgcaggta acgggcaacc aaggcgtaga
     5761 tccatccaag acttgactgt tactgggaca gaacctggtc aggtgagcag ccggtcatcc
     5821 agccctagtg tcagaatgat cactacctca ggaccaacct cagagaagcc agctcgcagt
     5881 cacccatgga cccctgatga ttccacagac accaatggct cagataactc catcccaatg
     5941 gcgtatctta cactggatca ccagctacag cctctagcgc cgtgcccaaa ctccaaagaa
     6001 tccatggcag tgttcgaaca gcactgtaaa atggcacagg agtatatgaa agttcaaacc
     6061 gaaatcgcat tgttactaca gagaaagcaa gaactagttg cagaattgga ccaggatgaa
     6121 aaggaccagc aaaatacatc tcgtctggta caggaacata aaaagctttt agatgaaaac
     6181 aaaagccttt ctacttatta ccagcaatgc aaaaaacaac tagaggtcat cagaagccag
     6241 cagcagaaac gacaaggcac ttcagaattc ggatccgact acaaggacga cgacgacaag
     6301 taaagcttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     6361 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     6421 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     6481 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     6541 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     6601 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6661 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6721 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6781 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6841 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     6901 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     6961 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7021 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     7081 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc a
//
