LOCUS       sequence-197332-        7063 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 7063)
  TITLE     NFkappaB EMSA Studies
REFERENCE   2  (bases 1 to 7063)
  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"
COMMENT     Sequence Label: RelB cFlag pcDNA3
FEATURES             Location/Qualifiers
     source          1..7063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2346..2369
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2382..2400)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2383..2400)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2420..2437)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2426..2650
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2696..3124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2783..2802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2993..3014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3133..3153)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3138..3467
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3318..3453
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3380..3399
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3534..4328
                     /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(3588..3607)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4198..4217
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4502..4623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4539..4558)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4593..4612
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4672..4688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4672..4688)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4685..4707)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4696..4712
                     /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(4720..4726,4727..4744,4745..4750))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4765..4786
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4903..4920)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5074..5662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5154..5173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5833..6624,6625..6693))
                     /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     6456..6475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6694..6798)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6873..6892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ccaccatgcc gagtcgccgc gctgccagag agtccgcgcc cgagctaggg gccttgggtt
      721 ccagtgacct ctcttccctg tcactaacgg tctccaggac cacagatgaa ttggaaatca
      781 tcgacgaata cattaaggag aacggctttg gcctggacgg gacacagctg agtgagatgc
      841 cgcgcctggt gccccgcggg cccgcctcac tgagcagcgt cacgctgggc cctgctgcac
      901 caccgcctcc ggccacgccg tcctggagct gcacactggg caggctggtg tcacccggcc
      961 cgtgcccacg gccgtacctg gtcatcacag agcagccaaa gcagcgtggc atgcgcttcc
     1021 gctacgagtg cgagggccgc tcggccggca gcatcctcgg ggagagcagc accgaagcca
     1081 gcaagaccct gcccgccatc gagcttcgag actgtggcgg gctgcgggag gtggaggtga
     1141 cggcgtgcct ggtgtggaag gactggccac accgggtaca cccacatagc ctcgtgggga
     1201 aagactgcac ggacggcgtc tgcagggtgc ggctgcggcc tcacgtcagc ccccggcaca
     1261 gctttaacaa cctgggcatc cagtgtgtta ggaagaagga aattgaagct gccattgagc
     1321 ggaagatcca gctgggaatt gacccctaca atgctggctc cctgaagaac catcaggagg
     1381 tcgacatgaa tgtcgtcagg atctgcttcc aggcctccta tcgggaccag cagggacatc
     1441 tgcaccgcat ggaccccatc ctctctgagc ctgtctacga caagaagtcc accaacacat
     1501 cggagctgcg gatttgccga atcaacaagg agagcgggcc gtgcacaggt ggtgaggagc
     1561 tgtacttgct ctgtgacaag gtgcaaaaag aggacatatc cgtggtgttc agcacagctt
     1621 cctgggaagg ccgtgccgac ttctctcaag ctgatgtgca ccggcagatc gccattgtgt
     1681 tcaaaacgcc accctacgag gacctggaga tctcagagcc cgtgactgtc aatgtgttct
     1741 tgcagcggct cacggatggg gtgtgcagcg agccgctgcc cttcacgtac ctgcctcggg
     1801 atcatgacag ctacggtgtg gacaagaagc gaaagcgggg actgcctgat gtccttggag
     1861 agttgagcag ctctgatcca catggaatcg agagcaaacg aaggaaaaag aaaccagtgt
     1921 tcttggacca cttcctgcct ggccacagct caggcctgtt cctcccacca tcggctctgc
     1981 agccggcaga ctctgatttc ttccctgctt ccatatccct tcctgggctg gagcctcctg
     2041 gtggacccga tctcctggac gatggctttg cctatgatcc ttctgccccc acgctcttca
     2101 ctatgttgga cctgctgccc ccagcaccac cacttgccag tgctgtggtg ggtagcgggg
     2161 gtgcaggggc caccgttgtg gagtcttctg gcccagagcc cctatcactg gactcttttg
     2221 cagcgccggg ccccggggat gttggtactg ctagccttgt gggcagcaac atgtttccca
     2281 accagtaccg agaggcagct ttcgggggtg gcctcctatc tccagggcct gaagccacgg
     2341 gatccgacta caaggacgac gatgacaagt aagaattcta ttctatagtg tcacctaaat
     2401 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2461 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2521 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2581 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2641 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2701 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2761 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2821 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2881 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2941 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3001 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3061 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3121 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3181 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3301 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3361 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3421 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3481 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3541 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3601 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3661 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3721 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3781 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3841 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3901 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3961 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4021 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4081 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4141 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4201 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4261 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4321 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4381 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4441 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4501 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4561 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4621 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     4681 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4741 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4801 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4861 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4921 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4981 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5581 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5641 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5701 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5881 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5941 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6001 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6061 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6121 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6181 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6241 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6301 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6361 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6421 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6481 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6541 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6601 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6661 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6721 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6781 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     6841 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6901 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     6961 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     7021 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gtt
//
