LOCUS       sequence-364499-        7338 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 7338)
  AUTHORS   Nakano H, Shindo M, Sakon S, Nishinaka S, Mihara M, Yagita H, 
            Okumura K
  TITLE     Differential regulation of IkappaB kinase alpha and beta by two 
            upstream kinases, NF-kappaB-inducing kinase and mitogen-activated 
            protein kinase/ERK kinase kinase-1.
  JOURNAL   Proc Natl Acad Sci U S A. 1998 Mar 31. 95(7):3537-42.
  PUBMED    9520401
REFERENCE   2  (bases 1 to 7338)
  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: "Proc Natl 
            Acad Sci U S A. 1998 Mar 31. 95(7):3537-42."
COMMENT     Sequence Label: pCR-Flag-IKKalpha
FEATURES             Location/Qualifiers
     source          1..7338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..289
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     246..266
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(349..1140,1141..1209))
                     /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     972..991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1210..1314)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1341..1796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1428..1447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1638..1659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        1808..2187
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2188..2391
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2342..2362
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2436..2455
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2436..2454
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2520..2543
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2553..2576
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     RBS             4537..4545
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(4845..4863)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4846..4863)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4883..4900)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4889..5113
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(5245..5833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5325..5344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6162..6209
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6234..6253
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(6441..7235)
                     /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(6441..7235)
                     /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(6552..6571)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7162..7181
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7270..7338,1..289))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7284..7338,1..81)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
       61 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
      121 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      181 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      241 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
      301 tttcaataaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      361 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      421 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      481 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      541 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      601 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      661 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      721 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      781 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      841 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      901 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      961 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1021 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1081 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1141 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1201 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1261 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1321 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1381 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1441 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1501 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1561 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1621 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1681 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1741 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacgc
     1801 gcgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     1861 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1921 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1981 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2041 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2101 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2161 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2221 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2281 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2341 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     2401 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
     2461 caagcttggt accgagctcg gatccactag taacggccgc cagtgtgctg gaattgatgg
     2521 actacaagga cgacgatgac aaggaattga tggactacaa ggacgacgat gacaaggaat
     2581 tcatggagcg gcccccgggg ctgcggccgg gcgcgggcgg cccctgggag atgcgggaac
     2641 ggcttggcac cggcggtttc gggaacgtca gtctgtacca gcaccgggaa cttgatctca
     2701 aaatagcaat taagtcttgt cgtttagagc taagttccaa aaacagagag cgatggtgcc
     2761 atgaaatcca gatcatgaaa aagttggacc atgcgaatgt tgtaaaggcc tgtgatgtcc
     2821 ctgaggaatt gaacttttta attaacgatg tgcctcttct ggcaatggag tactgttctg
     2881 gaggggacct ccggaagcta ctcaacaaac cagaaaattg ttgtggactt aaagaaagcc
     2941 agatactttc tttactgagt gacataggat ctgggatccg atatctgcat gaaaacaaaa
     3001 ttatacatcg agatctaaaa cctgaaaata tagttcttca agatgttggt gggaagacaa
     3061 tacataaaat aattgatttg ggttatgcca aagatgttga tcaaggaagt ctctgtacat
     3121 cttttgtggg aacattgcag tatttggccc cagagctctt tgaaaataag ccgtacacag
     3181 ccactgtgga ttattggagc tttgggacca tggtgtttga atgtattgct ggatataggc
     3241 cttttttgca tcatctgcag ccgtttacct ggcatgagaa gattaagaag aaagatccaa
     3301 agtgtatatt tgcatgtgaa gagatgactg gagaagttcg gtttagtagc catttacctc
     3361 agccaaacag cctttgtagt ttaatagtag agccaatgga aagctggctc caattgatgc
     3421 tgaattggga cccacagcag agagggggac ctattgatct tactttgaag cagccaagat
     3481 gttttgcatt aatggatcac attctcaatt taaagatagt gcacatccta aatatgactt
     3541 ctgcaaaaat catttctttt ctgttaccat gtgatgaaag tcttcattca ctacagtctc
     3601 gaattgagcg tgaaacagga ataaatacag gttctcagga gcttctgtca gagacaggga
     3661 tttctctgga tcctcggaaa ccagcctctc agtgtgttct agatggagtt agaggctgtg
     3721 atagctacat ggtttatttg tttgataaaa gtaagactgt atatgaagga ccatttgcat
     3781 ccagaagttt atctgattgt gtaaattata ttgtacaaga cagcaaaata caactgccaa
     3841 ttatacagct gcggaaagta tgggctgaag cagtgcacta cgtatctggg ctaaaggaag
     3901 actacagcag gctcttccag ggacaaagag cagcaatgtt aagtcttctt agatataatg
     3961 ctaacttgac aaaaatgaag aatactttga tctcagcatc acagcaactc aaagctaaat
     4021 tggagttttt tcgaaaaagc attcagcttg acttggagag atatagtgag cagatgactt
     4081 atgggatatc ttcagaaaaa atgttaaaag catggaaaga aatggaagaa aaggccattc
     4141 actattctga ggttggtgtc attggttatc ttgaggatca aattatgtct ttgcacactg
     4201 aaatcatgga gctgcagaag agcccctatg gacgacgcca gggagacttg atggagtctc
     4261 tggagcagcg tgccattgat ctctataagc agctaaagca cagacctcct gatcacttgt
     4321 acagcgacag cacagagatg gtgaagatca tcgtgcacac cgtgcagagt caggaccgtg
     4381 ttctcaagga gctgtttggt cacctgagca agttgttggg ctgcaagcag aagattattg
     4441 atctactccc caaggtggaa gtggccctca gtaacatcaa agaagctgac aatactgtca
     4501 tgtttatgca gggaaagagg cagaaagaaa tttggcacct ccttaaaatt gcctgtacac
     4561 agagttctgc ccgctctctt gtaggataca gtctagaagg cacagtaacc cctccagtat
     4621 cagcatggct gccccctaca ttagcagacc gtgaacatcc tctgacatgt gtggtaactc
     4681 ctcaagatgg agagacgtta gcacaaatga tagaagaaaa tctgaactgt cttggccatt
     4741 taagtactat tattcgtgaa gcaaatgagg accagagcag tagtttgatg agtcttgatt
     4801 ggagttggtt agcagaactc gagcatgcat ctagagggcc ctattctata gtgtcaccta
     4861 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4921 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4981 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     5041 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     5101 gtgggctcta tggcttctga ggcggaaaga accagtggcg gtaatacggt tatccacaga
     5161 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5221 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5281 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5341 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5401 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5461 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5521 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5581 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5641 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5701 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5761 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5821 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5881 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5941 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     6001 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     6061 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     6121 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     6181 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     6241 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     6301 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     6361 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     6421 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     6481 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     6541 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     6601 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     6661 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg
     6721 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     6781 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     6841 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     6901 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     6961 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     7021 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg
     7081 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     7141 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     7201 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     7261 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     7321 ctcagaggcc gaggcggc
//
