LOCUS       sequence-169429-        7193 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 7193)
  AUTHORS   Mercurio F, Zhu H, Murray BW, Shevchenko A, Bennett BL, Li J, Young 
            DB, Barbosa M, Mann M, Manning A, Rao A
  TITLE     IKK-1 and IKK-2: cytokine-activated IkappaB kinases essential for 
            NF-kappaB activation.
  JOURNAL   Science. 1997 Oct 31. 278(5339):860-6.
  PUBMED    9346484
REFERENCE   2  (bases 1 to 7193)
  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: "Science. 
            1997 Oct 31. 278(5339):860-6."
COMMENT     Sequence Label: IKK-2 K44M
FEATURES             Location/Qualifiers
     source          1..7193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1985..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2566..2588
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2580..2597
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2581..2597
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2758..3137
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3138..3341
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3292..3312
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3375..3398
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    5974..6596
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6173..6192)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6620..6640)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6625..6954
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6805..6940
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6992..7011)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6993..7011)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7025..7041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7025..7041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7038..7060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7049..7065
                     /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(7073..7079,7080..7097,7098..7103))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7118..7139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
     2461 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2521 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2581 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
     2641 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     2701 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
     2761 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2821 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2881 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2941 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3001 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3061 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
     3121 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
     3181 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3241 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
     3301 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     3361 attgatctac catggactac aaagacgatg acgacaagct tgcggccgca atgagctggt
     3421 caccttccct gacaacgcag acatgtgggg cctgggaaat gaaagagcgc cttgggacag
     3481 ggggatttgg aaatgtcatc cgatggcaca atcaggaaac aggtgagcag attgccatca
     3541 tgcagtgccg gcaggagctc agcccccgga accgagagcg gtggtgcctg gagatccaga
     3601 tcatgagaag gctgacccac cccaatgtgg tggctgcccg agatgtccct gaggggatgc
     3661 agaacttggc gcccaatgac ctgcccctgc tggccatgga gtactgccaa ggaggagatc
     3721 tccggaagta cctgaaccag tttgagaact gctgtggtct gcgggaaggt gccatcctca
     3781 ccttgctgag tgacattgcc tctgcgctta gataccttca tgaaaacaga atcatccatc
     3841 gggatctaaa gccagaaaac atcgtcctgc agcaaggaga acagaggtta atacacaaaa
     3901 ttattgacct aggatatgcc aaggagctgg atcagggcag tctttgcaca tcattcgtgg
     3961 ggaccctgca gtacctggcc ccagagctac tggagcagca gaagtacaca gtgaccgtcg
     4021 actactggag cttcggcacc ctggcctttg agtgcatcac gggcttccgg cccttcctcc
     4081 ccaactggca gcccgtgcag tggcattcaa aagtgcggca gaagagtgag gtggacattg
     4141 ttgttagcga agacttgaat ggaacggtga agttttcaag ctctttaccc taccccaata
     4201 atcttaacag tgtcctggct gagcgactgg agaagtggct gcaactgatg ctgatgtggc
     4261 acccccgaca gaggggcacg gatcccacgt atgggcccaa tggctgcttc aaggccctgg
     4321 atgacatctt aaacttaaag ttggttcata tcttgaacat ggtcacgggc accatccaca
     4381 cctaccctgt gacagaggat gagagtctgc agagcttgaa ggccagaatc caacaggaca
     4441 cgggcatccc agaggaggac caggagctgc tgcaggaagc gggcctggcg ttgatccccg
     4501 ataagcctgc cactcagtgt atttcagacg gcaagttaaa tgagggccac acattggaca
     4561 tggatcttgt ttttctcttt gacaacagta aaatcaccta tgagactcag atctccccac
     4621 ggccccaacc tgaaagtgtc agctgtatcc ttcaagagcc caagaggaat ctcgccttct
     4681 tccagctgag gaaggtgtgg ggccaggtct ggcacagcat ccagaccctg aaggaagatt
     4741 gcaaccggct gcagcaggga cagcgagccg ccatgatgaa tctcctccga aacaacagct
     4801 gcctctccaa aatgaagaat tccatggctt ccatgtctca gcagctcaag gccaagttgg
     4861 atttcttcaa aaccagcatc cagattgacc tggagaagta cagcgagcaa accgagtttg
     4921 ggatcacatc agataaactg ctgctggcct ggagggaaat ggagcaggct gtggagctct
     4981 gtgggcggga gaacgaagtg aaactcctgg tagaacggat gatggctctg cagaccgaca
     5041 ttgtggactt acagaggagc cccatgggcc ggaagcaggg gggaacgctg gacgacctag
     5101 aggagcaagc aagggagctg tacaggagac taagggaaaa acctcgagac cagcgaactg
     5161 agggtgacag tcaggaaatg gtacggctgc tgcttcaggc aattcagagc ttcgagaaga
     5221 aagtgcgagt gatctatacg cagctcagta aaactgtggt ttgcaagcag aaggcgctgg
     5281 aactgttgcc caaggtggaa gaggtggtga gcttaatgaa tgaggatgag aagactgttg
     5341 tccggctgca ggagaagcgg cagaaggagc tctggaatct cctgaagatt gcttgtagca
     5401 aggtccgtgg tcatgtcagt ggaagcccgg atagcatgaa tgcctctcga cttagccagc
     5461 ctgggcagct gatgtctcag ccctccacgg cctccaacag cttacctgag ccagccaaga
     5521 agagtgaaga actggtggct gaagcacata acctctgcac cctgctagaa aatgccatac
     5581 aggacactgt gagggaacaa gaccagagtt tcacggccct agactggagc tggttacaga
     5641 cggaagaaga agagcacagc tgcctggagc aggcctcatg atgtgggggg actcgacccc
     5701 ctgacatggg gcagcccata gcaggccttg tgcggtgggg ggactcgacc ccctgacatg
     5761 gggctgcctg gagcaggccg cgtgacgtgg ggctgcctgg ccgcggctct cacatggtgg
     5821 ttcctgctgc actgatggcc caggggtctc tggtatccag atggagctct cgcttcctca
     5881 gcagctgtga ctttcacccg tcgacttggg ctagagcggc cgcgaattca tcgatagatc
     5941 tgatatcggt accagtcgac tctagaggat cccgggtggc atccctgtga cccctcccca
     6001 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     6061 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     6121 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     6181 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     6241 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     6301 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     6361 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     6421 gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag
     6481 gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc
     6541 ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt
     6601 acgcggccag cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     6661 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     6721 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6781 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     6841 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     6901 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     6961 ctcgaggaac tgaaaaacca gaaagttaat tccctatagt gagtcgtatt aaattcgtaa
     7021 tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7081 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7141 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctg
//
