LOCUS       sequence-349392-        6501 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 6501)
  AUTHORS   Liao G, Zhang M, Harhaj EW, Sun SC
  TITLE     Regulation of the NF-kappaB-inducing kinase by tumor necrosis factor
            receptor-associated factor 3-induced degradation.
  JOURNAL   J Biol Chem. 2004 Jun 18;279(25):26243-50. Epub 2004 Apr 14.
  PUBMED    15084608
REFERENCE   2  (bases 1 to 6501)
  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: "2004-06-18"; volume: "279(25)"; pages: "26243-50. 
            Epub 2004 Apr 14"
COMMENT     Sequence Label: pcDNA-HA-hTRAF3 1-448
FEATURES             Location/Qualifiers
     source          1..6501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             321..347
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1752..1793
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1803..1820
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1843..1860)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1849..2073
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2119..2547
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2206..2225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2416..2437
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2556..2576)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2561..2890
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2741..2876
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2803..2822
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2938..2985
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3004..3402
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3560..3681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3597..3616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3651..3670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3730..3746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3730..3746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3743..3765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3754..3770
                     /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(3778..3784,3785..3802,3803..3808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3823..3844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3961..3978)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4132..4720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4212..4231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4891..5682,5683..5751))
                     /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     5514..5533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5752..5856)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5931..5950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6122..6501
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc
      301 ttggtaccga tatcatggcc tacccttatg atgtgccaga ttatgcctct cccgaattca
      361 ccatggagtc gagtaaaaag atggactctc ctggcgcgct gcagactaac ccgccgctaa
      421 agctgcacac tgaccgcagt gctgggacgc cagtttttgt ccctgaacaa ggaggttaca
      481 aggaaaagtt tgtgaagacc gtggaggaca agtacaagtg tgagaagtgc cacctggtgc
      541 tgtgcagccc gaagcagacc gagtgtgggc accgcttctg cgagagctgc atggcggccc
      601 tgctgagctc ttcaagtcca aaatgtacag cgtgtcaaga gagcatcgtt aaagataagg
      661 tgtttaagga taattgctgc aagagagaaa ttctggctct tcagatctat tgtcggaatg
      721 aaagcagagg ttgtgcagag cagttaatgc tgggacatct gctggtgcat ttaaaaaatg
      781 attgccattt tgaagaactt ccatgtgtgc gtcctgactg caaagaaaag gtcttgagga
      841 aagacctgcg agaccacgtg gagaaggcgt gtaaataccg ggaagccaca tgcagccact
      901 gcaagagtca ggttccgatg atcgcgctgc agaaacacga agacaccgac tgtccctgcg
      961 tggtggtgtc ctgccctcac aagtgcagcg tccagactct cctgaggagc gagttgagtg
     1021 cacacttgtc agagtgtgtc aatgccccca gcacctgtag ttttaagcgc tatggctgcg
     1081 tttttcaggg gacaaaccag cagatcaagg cccacgaggc cagctccgcc gtgcagcacg
     1141 tcaacctgct gaaggagtgg agcaactcgc tcgaaaagaa ggattccttg ttgcagaatg
     1201 aaagtgtaga aaaaaacaag agcatacaaa gtttgcacaa tcagatatgt agctttgaaa
     1261 ttgaaattga gagacaaaag gaaatgcttc gaaataatga atccaaaatc cttcatttac
     1321 agcgagtgat agacagccaa gcagagaaac tgaaggagct tgacaaggag atccggccct
     1381 tccggcagaa ctgggaggaa gcagacagca tgaagagcag cgtggagtcc ctccagaacc
     1441 gcgtgaccga gctggagagc gtggacaaga gcgcggggca agtggctcgg aacacaggcc
     1501 tgctggagtc ccagctgagc cggcatgacc agatgctggg tgtgcacgac atccgcctag
     1561 ccgacatgga cctgcgcttc caggtcctgg agaccgccag ctacaatgga gtgctcatct
     1621 ggaagattcg cgactacaag cggcggaagc aggaggccgt catggggaag accctgtccc
     1681 tttacagcca gcctttctac actggttaac tcgagaagct ttggacttct taatctagag
     1741 ggcccttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg
     1801 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     1861 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1921 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1981 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2041 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2101 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2161 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2221 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2281 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2341 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2401 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2461 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2521 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     2581 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2641 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2701 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2761 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2821 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2881 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     2941 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     3001 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     3061 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     3121 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     3181 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     3241 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     3301 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     3361 attcgtgaat tgctgccctc tggttatgtg tgggagggct aagcacttcg tggccgagga
     3421 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3481 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3541 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     3601 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3661 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     3721 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3781 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3841 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3901 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3961 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4021 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4081 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4141 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4201 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4261 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4321 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4381 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4441 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4501 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4561 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4621 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4681 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4741 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4801 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4861 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4921 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4981 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5041 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5101 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5161 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5221 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5281 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5341 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5401 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5461 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5521 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5581 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5641 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5701 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5761 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5821 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5881 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     5941 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     6001 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     6061 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     6121 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     6181 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6241 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6301 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6361 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6421 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6481 ttagtcatcg ctattaccat g
//
