LOCUS       Exported                6696 bp ds-DNA     circular SYN 16-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    FLAG-TRAF6-wt
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 Jun 16, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6696
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(34..53)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     244..265
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        414..793
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        794..997
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     948..968
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1042..1061
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1042..1060
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1126..1149
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1159..1182
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2809..2827)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2810..2827)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2847..2864)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2853..3077
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3209..3797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3289..3308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4126..4173
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4198..4217
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4405..5199)
                     /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(4516..4535)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5126..5145
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5234..5591)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5248..5383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5302..5321)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5548..5568
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(5651..6511)
                     /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     6274..6293
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6512..6616)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6643..402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
       61 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      121 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      181 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      241 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      301 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      361 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcgcgc gttgacattg
      421 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      481 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      541 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      601 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      661 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      721 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      781 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      841 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      901 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      961 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
     1021 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag cttggtaccg
     1081 agctcggatc cactagtaac ggccgccagt gtgctggaat tgatggacta caaggacgac
     1141 gatgacaagg aattgatgga ctacaaggac gacgatgaca aggaattcat gagtctctta
     1201 aactgtgaga acagctgcgg gtccagccag tcgtccagtg actgctgcgc tgccatggcc
     1261 gcctcctgca gcgctgcagt gaaagatgac agcgtgagtg gctctgccag caccgggaac
     1321 ctctccagct ccttcatgga ggagatccag ggctacgatg tggagtttga cccacctctg
     1381 gagagcaagt atgagtgtcc catttgcttg atggctttac gggaagcagt gcaaacacca
     1441 tgtggccaca ggttctgcaa agcctgcatc atcaaatcca taagggatgc agggcacaag
     1501 tgcccagttg acaatgaaat actgctggaa aatcaactgt ttcccgacaa ttttgcaaag
     1561 cgagagattc tttccctgac ggtaaagtgc ccaaataaag gctgtttgca aaagatggaa
     1621 ctgagacatc tcgaggatca tcaagtacat tgtgaatttg ctctagtgaa ttgtccccag
     1681 tgccaacgtc ctttccagaa gtgccaggtt aatacacaca ttattgagga ttgtcccagg
     1741 aggcaggttt cttgtgtaaa ctgtgctgtg tccatggcat atgaagagaa agagatccat
     1801 gatcaaagct gtcctctggc aaatatcatc tgtgaatact gtggtacaat cctcatcaga
     1861 gaacagatgc ctaatcatta tgatctggac tgcccaacag ctccaatccc ttgcacattc
     1921 agtgtttttg gctgtcatga aaagatgcag aggaatcact tggcacgaca cttgcaagag
     1981 aatacccagt tgcacatgag actgttggcc caggctgttc ataatgttaa ccttgctttg
     2041 cgtccgtgcg atgccgcctc tccatcccgg ggatgtcgtc cagaggaccc aaattatgag
     2101 gaaactatca aacagttgga gagtcgccta gtaagacagg accatcagat ccgggagctg
     2161 actgccaaaa tggaaactca gagtatgtac gtgggcgagc tcaaacggac cattcggacc
     2221 ctggaggaca aggttgccga aatggaagca cagcagtgta acgggatcta catttggaag
     2281 attggcaact ttgggatgca cttgaaatcc caagaagagg aaagacctgt tgtcatccat
     2341 agccctggat tctacacagg cagacctggg tacaagctgt gcatgcgcct gcatcttcag
     2401 ttaccgacag ctcagcgctg tgcaaactat atatcccttt ttgtccacac aatgcaagga
     2461 gaatatgaca gccacctccc ctggcccttc cagggtacaa tacgccttac aattctcgac
     2521 cagtctgaag cacttataag gcaaaaccac gaagaggtca tggacgccaa accagaactg
     2581 cttgcctttc agcgacccac aatcccacgg aaccccaaag gttttggcta tgtaacattt
     2641 atgcacctgg aagccttaag acagggaacc ttcattaagg atgatacatt actagtgcgc
     2701 tgtgaagtct ctacccgctt tgacatgggt ggccttcgga aggagggttt ccagccacga
     2761 agtactgatg cgggggtgta ggtcgagcat gcatctagag ggccctattc tatagtgtca
     2821 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2881 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2941 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3001 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3061 tgcggtgggc tctatggctt ctgaggcgga aagaaccagt ggcggtaata cggttatcca
     3121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3781 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3841 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3901 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taacctgagg
     3961 ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca cccgaacttg
     4021 gggggtgggg tggggaaaag gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg
     4081 gggtatcgac agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa
     4141 caccgtgcgt tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg
     4201 tctccttccg tgtttcagtt agcctccccc tagggtgggc gaagaactcc agcatgagat
     4261 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     4321 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
     4381 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     4441 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     4501 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     4561 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     4621 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag
     4681 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     4741 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     4801 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     4861 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     4921 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     4981 cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga
     5041 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     5101 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     5161 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     5221 ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     5281 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     5341 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     5401 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     5461 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
     5521 ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac agctggttct
     5581 ttccgcctca ggactcttcc tttttcaata aatcaatcta aagtatatat gagtaaactt
     5641 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5701 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5761 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5821 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5881 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5941 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6001 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6061 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6121 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6181 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6241 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6301 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6361 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6421 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6481 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6541 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6601 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     6661 cattaagcgc ggcgggtgtg gtggttacgc gcagcg
//
