LOCUS       sequence-134979-        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 WT
FEATURES             Location/Qualifiers
     source          1..7193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1933..2388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2020..2039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2230..2251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2514..2536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2528..2545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2706..3085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3086..3289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3240..3260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3323..3346
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    5922..6544
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6121..6140)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6568..6588)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6573..6902
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6753..6888
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6940..6959)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6941..6959)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6973..6989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6973..6989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6986..7008)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6997..7013
                     /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(7021..7027,7028..7045,7046..7051))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7066..7087
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1981 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2221 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2281 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2341 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
     2581 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     2641 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     2701 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     2761 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2821 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2881 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2941 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     3001 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     3061 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     3121 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     3181 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     3241 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     3301 ccgtcagaat tgatctacca tggactacaa agacgatgac gacaagcttg cggccgcaat
     3361 gagctggtca ccttccctga caacgcagac atgtggggcc tgggaaatga aagagcgcct
     3421 tgggacaggg ggatttggaa atgtcatccg atggcacaat caggaaacag gtgagcagat
     3481 tgccatcaag cagtgccggc aggagctcag cccccggaac cgagagcggt ggtgcctgga
     3541 gatccagatc atgagaaggc tgacccaccc caatgtggtg gctgcccgag atgtccctga
     3601 ggggatgcag aacttggcgc ccaatgacct gcccctgctg gccatggagt actgccaagg
     3661 aggagatctc cggaagtacc tgaaccagtt tgagaactgc tgtggtctgc gggaaggtgc
     3721 catcctcacc ttgctgagtg acattgcctc tgcgcttaga taccttcatg aaaacagaat
     3781 catccatcgg gatctaaagc cagaaaacat cgtcctgcag caaggagaac agaggttaat
     3841 acacaaaatt attgacctag gatatgccaa ggagctggat cagggcagtc tttgcacatc
     3901 attcgtgggg accctgcagt acctggcccc agagctactg gagcagcaga agtacacagt
     3961 gaccgtcgac tactggagct tcggcaccct ggcctttgag tgcatcacgg gcttccggcc
     4021 cttcctcccc aactggcagc ccgtgcagtg gcattcaaaa gtgcggcaga agagtgaggt
     4081 ggacattgtt gttagcgaag acttgaatgg aacggtgaag ttttcaagct ctttacccta
     4141 ccccaataat cttaacagtg tcctggctga gcgactggag aagtggctgc aactgatgct
     4201 gatgtggcac ccccgacaga ggggcacgga tcccacgtat gggcccaatg gctgcttcaa
     4261 ggccctggat gacatcttaa acttaaagtt ggttcatatc ttgaacatgg tcacgggcac
     4321 catccacacc taccctgtga cagaggatga gagtctgcag agcttgaagg ccagaatcca
     4381 acaggacacg ggcatcccag aggaggacca ggagctgctg caggaagcgg gcctggcgtt
     4441 gatccccgat aagcctgcca ctcagtgtat ttcagacggc aagttaaatg agggccacac
     4501 attggacatg gatcttgttt ttctctttga caacagtaaa atcacctatg agactcagat
     4561 ctccccacgg ccccaacctg aaagtgtcag ctgtatcctt caagagccca agaggaatct
     4621 cgccttcttc cagctgagga aggtgtgggg ccaggtctgg cacagcatcc agaccctgaa
     4681 ggaagattgc aaccggctgc agcagggaca gcgagccgcc atgatgaatc tcctccgaaa
     4741 caacagctgc ctctccaaaa tgaagaattc catggcttcc atgtctcagc agctcaaggc
     4801 caagttggat ttcttcaaaa ccagcatcca gattgacctg gagaagtaca gcgagcaaac
     4861 cgagtttggg atcacatcag ataaactgct gctggcctgg agggaaatgg agcaggctgt
     4921 ggagctctgt gggcgggaga acgaagtgaa actcctggta gaacggatga tggctctgca
     4981 gaccgacatt gtggacttac agaggagccc catgggccgg aagcaggggg gaacgctgga
     5041 cgacctagag gagcaagcaa gggagctgta caggagacta agggaaaaac ctcgagacca
     5101 gcgaactgag ggtgacagtc aggaaatggt acggctgctg cttcaggcaa ttcagagctt
     5161 cgagaagaaa gtgcgagtga tctatacgca gctcagtaaa actgtggttt gcaagcagaa
     5221 ggcgctggaa ctgttgccca aggtggaaga ggtggtgagc ttaatgaatg aggatgagaa
     5281 gactgttgtc cggctgcagg agaagcggca gaaggagctc tggaatctcc tgaagattgc
     5341 ttgtagcaag gtccgtggtc ctgtcagtgg aagcccggat agcatgaatg cctctcgact
     5401 tagccagcct gggcagctga tgtctcagcc ctccacggcc tccaacagct tacctgagcc
     5461 agccaagaag agtgaagaac tggtggctga agcacataac ctctgcaccc tgctagaaaa
     5521 tgccatacag gacactgtga gggaacaaga ccagagtttc acggccctag actggagctg
     5581 gttacagacg gaagaagaag agcacagctg cctggagcag gcctcatgat gtggggggac
     5641 tcgaccccct gacatggggc agcccatagc aggccttgtg cggtgggggg actcgacccc
     5701 ctgacatggg gctgcctgga gcaggccgcg tgacgtgggg ctgcctggcc gcggctctca
     5761 catggtggtt cctgctgcac tgatggccca ggggtctctg gtatccagat ggagctctcg
     5821 cttcctcagc agctgtgact ttcacccgtc gacttgggct agagcggccg cgaattcatc
     5881 gatagatctg atatcggtac cagtcgactc tagaggatcc cgggtggcat ccctgtgacc
     5941 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     6001 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     6061 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     6121 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     6181 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     6241 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     6301 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     6361 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     6421 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     6481 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     6541 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     6601 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     6661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6721 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     6781 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6841 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     6901 aaaagctcct cgaggaactg aaaaaccaga aagttaattc cctatagtga gtcgtattaa
     6961 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7021 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7081 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7141 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gct
//
