LOCUS       sequence-364555-        6696 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 6696)
  AUTHORS   Zhong J, Kyriakis JM
  TITLE     Germinal center kinase is required for optimal Jun N-terminal kinase
            activation by Toll-like receptor agonists and is regulated by the 
            ubiquitin proteasome system and agonist-induced, TRAF6-dependent 
            stabilization.
  JOURNAL   Mol Cell Biol. 2004 Oct . 24(20):9165-75.
  PUBMED    15456887
REFERENCE   2  (bases 1 to 6696)
  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: "Mol Cell 
            Biol. 2004 Oct . 24(20):9165-75."
COMMENT     Sequence Label: FLAG-TRAF6-wt
FEATURES             Location/Qualifiers
     source          1..6696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        43..234
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     191..211
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(294..1085,1086..1154))
                     /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     917..936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1155..1259)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1286..1741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1373..1392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1583..1604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        1753..2132
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2133..2336
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2287..2307
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2381..2400
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2381..2399
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2465..2488
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2498..2521
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(4148..4166)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4149..4166)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4186..4203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4192..4416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4548..5136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4628..4647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5465..5512
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5537..5556
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5744..6538)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5744..6538)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5855..5874)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6465..6484
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6573..6696,1..234))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6587..6696,1..26)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
       61 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      121 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
      181 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
      241 tcctttttca ataaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
      301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
      361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
      421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
      481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
      541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
      601 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
      661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
      721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
      781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
      841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
      901 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1141 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1201 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     1261 acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1321 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1381 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1441 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1501 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1561 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1621 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1681 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1741 tacgcgcgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     1801 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     1861 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     1921 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     1981 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     2041 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     2101 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     2161 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     2221 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     2281 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
     2341 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
     2401 agacccaagc ttggtaccga gctcggatcc actagtaacg gccgccagtg tgctggaatt
     2461 gatggactac aaggacgacg atgacaagga attgatggac tacaaggacg acgatgacaa
     2521 ggaattcatg agtctcttaa actgtgagaa cagctgcggg tccagccagt cgtccagtga
     2581 ctgctgcgct gccatggccg cctcctgcag cgctgcagtg aaagatgaca gcgtgagtgg
     2641 ctctgccagc accgggaacc tctccagctc cttcatggag gagatccagg gctacgatgt
     2701 ggagtttgac ccacctctgg agagcaagta tgagtgtccc atttgcttga tggctttacg
     2761 ggaagcagtg caaacaccat gtggccacag gttctgcaaa gcctgcatca tcaaatccat
     2821 aagggatgca gggcacaagt gcccagttga caatgaaata ctgctggaaa atcaactgtt
     2881 tcccgacaat tttgcaaagc gagagattct ttccctgacg gtaaagtgcc caaataaagg
     2941 ctgtttgcaa aagatggaac tgagacatct cgaggatcat caagtacatt gtgaatttgc
     3001 tctagtgaat tgtccccagt gccaacgtcc tttccagaag tgccaggtta atacacacat
     3061 tattgaggat tgtcccagga ggcaggtttc ttgtgtaaac tgtgctgtgt ccatggcata
     3121 tgaagagaaa gagatccatg atcaaagctg tcctctggca aatatcatct gtgaatactg
     3181 tggtacaatc ctcatcagag aacagatgcc taatcattat gatctggact gcccaacagc
     3241 tccaatccct tgcacattca gtgtttttgg ctgtcatgaa aagatgcaga ggaatcactt
     3301 ggcacgacac ttgcaagaga atacccagtt gcacatgaga ctgttggccc aggctgttca
     3361 taatgttaac cttgctttgc gtccgtgcga tgccgcctct ccatcccggg gatgtcgtcc
     3421 agaggaccca aattatgagg aaactatcaa acagttggag agtcgcctag taagacagga
     3481 ccatcagatc cgggagctga ctgccaaaat ggaaactcag agtatgtacg tgggcgagct
     3541 caaacggacc attcggaccc tggaggacaa ggttgccgaa atggaagcac agcagtgtaa
     3601 cgggatctac atttggaaga ttggcaactt tgggatgcac ttgaaatccc aagaagagga
     3661 aagacctgtt gtcatccata gccctggatt ctacacaggc agacctgggt acaagctgtg
     3721 catgcgcctg catcttcagt taccgacagc tcagcgctgt gcaaactata tatccctttt
     3781 tgtccacaca atgcaaggag aatatgacag ccacctcccc tggcccttcc agggtacaat
     3841 acgccttaca attctcgacc agtctgaagc acttataagg caaaaccacg aagaggtcat
     3901 ggacgccaaa ccagaactgc ttgcctttca gcgacccaca atcccacgga accccaaagg
     3961 ttttggctat gtaacattta tgcacctgga agccttaaga cagggaacct tcattaagga
     4021 tgatacatta ctagtgcgct gtgaagtctc tacccgcttt gacatgggtg gccttcggaa
     4081 ggagggtttc cagccacgaa gtactgatgc gggggtgtag gtcgagcatg catctagagg
     4141 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     4201 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4261 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4321 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4381 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagtg
     4441 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4501 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4561 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4621 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4681 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4741 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4801 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4861 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4921 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4981 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5041 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5101 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5161 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5221 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5281 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     5341 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     5401 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     5461 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     5521 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     5581 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     5641 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     5701 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     5761 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     5821 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     5881 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     5941 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     6001 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     6061 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     6121 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     6181 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     6241 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     6301 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     6361 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     6421 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     6481 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     6541 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     6601 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     6661 aaaaaaatta gtcagccatg gggcggagaa tgggcg
//
