LOCUS       sequence-156218-        5316 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 5316)
  AUTHORS   Savinova OV, Hoffmann A, Ghosh G
  TITLE     The Nfkb1 and Nfkb2 proteins p105 and p100 function as the core of 
            high-molecular-weight heterogeneous complexes.
  JOURNAL   Mol Cell. 2009 Jun 12;34(5):591-602. doi: 
            10.1016/j.molcel.2009.04.033.
  PUBMED    19524538
REFERENCE   2  (bases 1 to 5316)
  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"; doi: 
            "10.1016/j.molcel.2009.04"; journalName: "Mol Cell"; date: 
            "2009-06-12- 12"; volume: "34"; issue: "5"; pages: "591-602"
COMMENT     Sequence Label: Flag-p50(1-435)
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             616..639
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2104..2225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2141..2160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2195..2214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2232..2687)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2369..2390)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2581..2600
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2714..2818
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2820..3177
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2820..3011
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2843..2863)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3028..3163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3212..4006
                     /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             3212..4006
                     /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(3266..3285)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3876..3895
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4194..4213)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4238..4285
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4614..5202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5103..5122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggacta caaagacgat gacgacaaga gatctcgagc tcaagcttcg
      661 aattctatgg cagacgatga tccctacgga actgggcaaa tgtttcattt gaacactgct
      721 ttgactcact caatatttaa tgcagaatta tattcaccag aaataccact gtcaacagat
      781 ggcccatacc ttcaaatatt agagcaacca aaacagaggg gatttcgatt ccgctatgtg
      841 tgtgaaggcc catcacacgg agggcttccg ggagcctcta gtgagaagaa caagaaatcc
      901 tacccacagg tcaaaatttg caactatgtg gggcctgcaa aggttatcgt tcagttggtc
      961 acaaatggaa aaaacatcca cctgcacgcc cacagcctgg tgggcaagca ctgtgaggac
     1021 ggggtatgca ccgtaacagc aggacccaag gacatggtgg ttggctttgc aaacctggga
     1081 atacttcatg tgactaagaa aaaggtattt gaaacactgg aagcacggat gacagaggcg
     1141 tgtattaggg gctataatcc tggacttctg gtgcattctg accttgccta tctacaagca
     1201 gaaggcggag gagaccggca actcacagac agagagaagg agatcatccg ccaggcagcc
     1261 gtgcagcaga ccaaggagat ggacctgagc gtggtgcgcc tcatgttcac agccttcctc
     1321 cctgacagca ctggcagctt cactcggaga ctggagcctg tggtgtcaga cgccatctat
     1381 gatagcaaag ccccgaatgc atccaacctg aaaatcgtga gaatggacag aacagcagga
     1441 tgtgtgacgg gaggggagga gatttacctt ctctgtgaca aggttcagaa agatgacatc
     1501 cagattcggt tttatgaaga ggaagaaaat ggcggagttt gggaaggatt tggggacttt
     1561 tcccccacgg atgttcatag acagtttgcc attgtcttca aaacgccaaa gtataaggat
     1621 gtcaacatta caaagccagc ttccgtgttt gttcagcttc ggaggaaatc agacctggaa
     1681 actagtgaac cgaaaccctt tctctactac cctgaaatca atgacaaaga ggaagtgcaa
     1741 aggaaacgcc agaagcttat gccgaacttc tcggacagct tcggcggcgg cagtggagcg
     1801 ggagccggtg gtggaggcat gttcggtagt ggcggtggcg gagggagtac cggaagccct
     1861 ggcccagggt atggctactc gaactacgga tttcctccct acggtgggat tacattccat
     1921 cccggagtca cgaaatccaa cgcaggggtc acccatggca ccataaacac ctagggatcc
     1981 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     2041 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2101 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2161 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2221 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2281 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2341 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2401 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2461 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2521 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2581 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2641 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     2701 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2761 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     2821 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2941 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3001 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3061 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3121 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3181 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3241 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3301 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3361 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3421 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3481 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3541 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3601 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3661 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3721 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3781 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3841 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3901 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3961 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4021 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4081 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4141 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4201 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4261 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4321 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4381 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4441 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4501 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4561 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4621 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4681 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4741 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4801 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4861 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4921 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4981 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5041 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5101 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5161 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5221 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5281 cctgattctg tggataaccg tattaccgcc atgcat
//
