LOCUS       sequence-495683-        7653 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7653)
  AUTHORS   Geleziunas R, Ferrell S, Lin X, Mu Y, Cunningham ET, Grant M, 
            Connelly MA, Hambor JE, Marcu KB, Greene WC
  TITLE     Human T-cell leukemia virus type 1 Tax induction of NF-kappaB 
            involves activation of the IkappaB kinase alpha (IKKalpha) and 
            IKKbeta cellular kinases.
  JOURNAL   Mol Cell Biol. 1998 Sep . 18(9):5157-65.
  PUBMED    9710600
REFERENCE   2  (bases 1 to 7653)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol. 1998 Sep . 18(9):5157-65."
COMMENT     Sequence Label: pcDNA-Ikkb-FLAG(K44A)
FEATURES             Location/Qualifiers
     source          1..7653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..554
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        555..758
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     709..729
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     803..822
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        803..821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3132..3155
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3184..3201)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3190..3414
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3460..3888
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3547..3566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3757..3778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3897..3917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3902..4231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4082..4217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4144..4163
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4298..5092
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4352..4371)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4962..4981
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5266..5387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5303..5322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5357..5376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5436..5452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5436..5452)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5449..5471)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5460..5476
                     /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(5484..5490,5491..5508,5509..5514))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5529..5550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5667..5684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5838..6426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5918..5937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6597..7388,7389..7457))
                     /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     7220..7239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7458..7562)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(7637..7653,1..3))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
       61 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      121 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      181 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      241 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      301 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      361 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      421 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      481 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      541 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      601 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      661 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      721 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      781 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      841 gtttaaacgg gccctctaga agtatgagct ggtcaccttc cctgacaacg cagacatgtg
      901 gggcctggga aatgaaagag cgccttggga cagggggatt tggaaatgtc atccgatggc
      961 acaatcagga aacaggtgag cagattgcca tcgcgcagtg ccggcaggag ctcagccccc
     1021 ggaaccgaga gcggtggtgc ctggagatcc agatcatgag aaggctgacc caccccaatg
     1081 tggtggctgc ccgagatgtc cctgagggga tgcagaactt ggcgcccaat gacctgcccc
     1141 tgctggccat ggagtactgc caaggaggag atctccggaa gtacctgaac cagtttgaga
     1201 actgctgtgg tctgcgggaa ggtgccatcc tcaccttgct gagtgacatt gcctctgcgc
     1261 ttagatacct tcatgaaaac agaatcatcc atcgggatct aaagccagaa aacatcgtcc
     1321 tgcagcaagg agaacagagg ttaatacaca aaattattga cctaggatat gccaaggagc
     1381 tggatcaggg cagtctttgc acatcattcg tggggaccct gcagtacctg gccccagagc
     1441 tactggagca gcagaagtac acagtgaccg tcgactactg gagcttcggc accctggcct
     1501 ttgagtgcat cacgggcttc cggcccttcc tccccaactg gcagcccgtg cagtggcatt
     1561 caaaagtgcg gcagaagagt gaggtggaca ttgttgttag cgaagacttg aatggaacgg
     1621 tgaagttttc aagctcttta ccctacccca ataatcttaa cagtgtcctg gctgagcgac
     1681 tggagaagtg gctgcaactg atgctgatgt ggcacccccg acagaggggc acggatccca
     1741 cgtatgggcc caatggctgc ttcaaggccc tggatgacat cttaaactta aagctggttc
     1801 atatcttgaa catggtcacg ggcaccatcc acacctaccc tgtgacagag gatgagagtc
     1861 tgcagagctt gaaggccaga atccaacagg acacgggcat cccagaggag gaccaggagc
     1921 tgctgcagga agcgggcctg gcgttgatcc ccgataagcc tgccactcag tgtatttcag
     1981 acggcaagtt aaatgagggc cacacattgg acatggatct tgtttttctc tttgacaaca
     2041 gtaaaatcac ctatgagact cagatctccc cacggcccca acctgaaagt gtcagctgta
     2101 tccttcaaga gcccaagagg aatctcgcct tcttccagct gaggaaggtg tggggccagg
     2161 tctggcacag catccagacc ctgaaggaag attgcaaccg gctgcagcag ggacagcgag
     2221 ccgccatgat gaatctcctc cgaaacaaca gctgcctctc caaaatgaag aattccatgg
     2281 cttccatgtc tcagcagctc aaggccaagt tggatttctt caaaaccagc atccagattg
     2341 acctggagaa gtacagcgag caaaccgagt ttgggatcac atcagataaa ctgctgctgg
     2401 cctggaggga aatggagcag gctgtggagc tctgtgggcg ggagaacgaa gtgaaactcc
     2461 tggtagaacg gatgatggct ctgcagaccg acattgtgga cttacagagg agccccatgg
     2521 gccggaagca ggggggaacg ctggacgacc tagaggagca agcaagggag ctgtacagga
     2581 gactaaggga aaaacctcga gaccagcgaa ctgagggtga cagtcaggaa atggtacggc
     2641 tgctgcttca ggcaattcag agcttcgaga agaaagtgcg agtgatctat acgcagctca
     2701 gtaaaactgt ggtttgcaag cagaaggcgc tggaactgtt gcccaaggtg gaagaggtgg
     2761 tgagcttaat gaatgaggat gagaagactg ttgtccggct gcaggagaag cggcagaagg
     2821 agctctggaa tctcctgaag attgcttgta gcaaggtccg tggtcctgtc agtggaagcc
     2881 cggatagcat gaatgcctct cgacttagcc agcctgggca gctgatgtct cagccctcca
     2941 cggcctccaa cagcttacct gagccagcca agaagagtga agaactggtg gctgaagcac
     3001 ataacctctg caccctgcta gaaaatgcca tacaggacac tgtgagggaa caagaccaga
     3061 gtttcacggc cctagactgg agctggttac agacggaaga agaagagcac agctgcctgg
     3121 agcaggcctc agactacaag gacgacgacg acaagtgaag cttaagttta aaccgctgat
     3181 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3241 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3301 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3361 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3421 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3481 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3541 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3601 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3661 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3721 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3781 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3841 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3901 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3961 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4081 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4141 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4201 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4261 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4321 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4381 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4441 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4501 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4561 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4621 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4681 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4741 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4801 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4861 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4921 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4981 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5041 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5101 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5161 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5221 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5281 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5341 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5401 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5461 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5521 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5581 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5641 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5701 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5761 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5821 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5881 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5941 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6001 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6061 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6121 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6181 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6241 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6301 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6361 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6421 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6481 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6541 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6601 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6661 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6721 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6781 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6841 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6901 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6961 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7021 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7081 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7141 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7201 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7261 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7321 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7381 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7441 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7501 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7561 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7621 tatggtgcac tctcagtaca atctgctctg atg
//
