LOCUS       sequence-169435-        6761 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 6761)
  TITLE     NFkappaB EMSA Studies
REFERENCE   2  (bases 1 to 6761)
  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: p50 cFlag pcDNA3
FEATURES             Location/Qualifiers
     source          1..6761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1294..1317
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1330..1348)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1331..1348)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1368..1385)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1374..1598
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1644..2072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1731..1750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1941..1962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2081..2101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2171..2487
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2338..2473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2400..2419
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2554..3348
                     /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(2608..2627)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3218..3237
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3522..3643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3559..3578)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3613..3632
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3692..3708)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3692..3708)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3705..3727)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3716..3732
                     /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(3740..3746,3747..3764,3765..3770))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3785..3806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3923..3940)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4094..4682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4174..4193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4853..5644,5645..5713))
                     /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     5476..5495
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5714..5818)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5893..5912)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6084..6463
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6464..6667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6618..6638
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6712..6731
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6712..6730
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      join(6756..6761,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggcagacg atgatcccta cggaactggg caaatgtttc atttgaacac tgctttgact
       61 cactcaatat ttaatgcaga attatattca ccagaaatac cactgtcaac agatggccca
      121 taccttcaaa tattagagca accaaaacag aggggatttc gattccgcta tgtgtgtgaa
      181 ggcccatcac acggagggct tccgggagcc tctagtgaga agaacaagaa atcctaccca
      241 caggtcaaaa tttgcaacta tgtggggcct gcaaaggtta tcgttcagtt ggtcacaaat
      301 ggaaaaaaca tccacctgca cgcccacagc ctggtgggca agcactgtga ggacggggta
      361 tgcaccgtaa cagcaggacc caaggacatg gtggttggct ttgcaaacct gggaatactt
      421 catgtgacta agaaaaaggt atttgaaaca ctggaagcac ggatgacaga ggcgtgtatt
      481 aggggctata atcctggact tctggtgcat tctgaccttg cctatctaca agcagaaggc
      541 ggaggagacc ggcaactcac agacagagag aaggagatca tccgccaggc agccgtgcag
      601 cagaccaagg agatggacct gagcgtggtg cgcctcatgt tcacagcctt cctccctgac
      661 agcactggca gcttcactcg gagactggag cctgtggtgt cagacgccat ctatgatagc
      721 aaagccccga atgcatccaa cctgaaaatc gtgagaatgg acagaacagc aggatgtgtg
      781 acgggagggg aggagattta ccttctctgt gacaaggttc agaaagatga catccagatt
      841 cggttttatg aagaggaaga aaatggcgga gtttgggaag gatttgggga cttttccccc
      901 acggatgttc atagacagtt tgccattgtc ttcaaaacgc caaagtataa ggatgtcaac
      961 attacaaagc cagcttccgt gtttgttcag cttcggagga aatcagacct ggaaactagt
     1021 gaaccgaaac cctttctcta ctaccctgaa atcaaagaca aagaggaagt gcaaaggaaa
     1081 cgccagaagc ttatgccgaa cttctcggac agcttcggcg gcggcagtgg agcgggagcc
     1141 ggtggtggag gcatgttcgg tagtggcggt ggcggaggga gtaccggaag ccctggccca
     1201 gggtatggct actcgaacta cggatttcct ccctacggtg ggattacatt ccatcccgga
     1261 gtcacgaaat ccaacgcagg ggtcaccgga tccgactaca aggacgacga tgacaagtaa
     1321 gaattctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     1381 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1441 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1501 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1561 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     1621 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1681 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1741 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1801 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     1861 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     1921 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     1981 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2041 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     2101 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2161 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2221 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2281 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     2341 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2401 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     2461 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     2521 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     2581 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2641 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2701 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     2761 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2821 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     2881 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     2941 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3001 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3061 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     3121 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3181 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3241 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3301 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3361 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3421 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3481 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3541 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3601 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     3661 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3721 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3781 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3841 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3901 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3961 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4021 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4081 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4141 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4201 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4261 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4321 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4381 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4441 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4501 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4561 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4621 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4681 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4741 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4801 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4861 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4921 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4981 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5041 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5101 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5161 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5221 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5281 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5341 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5401 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5461 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5521 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5581 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5641 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5701 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5761 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5821 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     5881 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     5941 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     6001 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     6061 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     6121 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     6181 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6241 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6301 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6361 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6421 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     6481 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6541 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     6601 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     6661 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     6721 tcactatagg gagacccaag cttatctaga ctcgagccac c
//
