LOCUS       sequence-354448-        5554 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 5554)
  AUTHORS   Wu CJ, Conze DB, Li T, Srinivasula SM, Ashwell JD
  TITLE     NEMO is a sensor of Lys 63-linked polyubiquitination and functions 
            in NF-kappaB activation.
  JOURNAL   Nat Cell Biol. 2006 Apr . 8(4):398-406.
  PUBMED    16547522
REFERENCE   2  (bases 1 to 5554)
  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: "Nat Cell 
            Biol. 2006 Apr . 8(4):398-406."
COMMENT     Sequence Label: pCMVTAG-NEMO
FEATURES             Location/Qualifiers
     source          1..5554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1359..1382)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1382..1391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1426..1444)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(1490..1693)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1519..1539)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1694..1997
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2172..2760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2252..2271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3089..3136
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3161..3180
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3368..4162)
                     /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(3368..4162)
                     /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(3479..3498)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4089..4108
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4197..4554)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4211..4346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4363..4554
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4511..4531
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4556..4660)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4687..5142
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4774..4793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4984..5005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5149..5270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5160..5179)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5214..5233
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     join(5544..5554,1..9)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5544..5554,1..8)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 actatagggc gaattgggta cacttacctg gtaccttaat taattaaggt accgggcccc
       61 ccctcgagct actcaatgca ctccatgaca tgtatctgca gggtgtccat atcaggggcc
      121 tgatactggc acttgggaca gcagaagtca ggtggctcct cgggggggct cctcctctgg
      181 ctgggcaggg ccaggggaga ggagaggtag gcaggggcgg ggggcaaggg ggcctgggag
      241 acctcgacat gccgcttcct catgtcctcg atcctggccg actcctgaca gctggccttc
      301 agtttgctgt actccctctg cagctgctcc agctgctcct gcaggagctc cttcttctcg
      361 gccagcttct cccgggcctg cctctcagcc tggaagtccg ccttgtagat atccgcctgg
      421 gccttcagca ccggaacggt ctccatcaca atcttgtgct gctcggcctc ctccttcagc
      481 ttatcgatca cctcctgttt ggccaccagg gcctcctcgg cctgctggag ctgctgtttg
      541 agatcttcca gctgcattcc tcgcttccgc tcactgccca ccacgctgct cttgatgtgg
      601 ttgtcgtatt cttggaagag ctggtgatag gccacctgca actgggccag cttcctcttc
      661 tcctccgagg cggcctggcg ctccatgcgg agcgcggcct ccacgctctg gccctgcatg
      721 cgcagctggt ccacctgcac gctgtgctgc tgctgcagcg cctcgcgctc actctccagc
      781 tgccgcgcct gctcgctggc cgcccgggcc cgaccctcca gagcctggca ttccttagtg
      841 gcagcctcca agcgactctg gctctcctgc agctccccga gcaaggacgt cacctgggct
      901 ttcacagagg ccttgtcctc agccatctgc tgctggcatc tcttcaggtg ctccacctcc
      961 cgcagagcct gctccttctg cctcttcaga tcgagcttct ccaggccgag tctctccacc
     1021 agtttcctgg cctcctggaa cttgcacatg aggaactcct tctcctccct ctggctggct
     1081 tggaaatgca gaagctcctc gcagcgctcc cgcagaatct ggttgctctg ccggatggca
     1141 tctcggagct cttgattctc ctccaggcag cgctggaggg tctcaggagc gccctgttct
     1201 gaaggcaggt gcagcatggc tggcttcccc agaggagact cttcgcccag tacgtcctga
     1261 tctgctgccg ggccaccact gggctgcacc atctcacaca gttggctctt ccagaggtgc
     1321 ctattgaatt cctgcagccc gggggatccg cccgggctct tatcgtcgtc atccttgtaa
     1381 tccatggtgg cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     1441 aatttcgagc ttggcgtaat cgctagcgga tctgacggtt cactaaacca gctctgctta
     1501 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     1561 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     1621 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     1681 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     1741 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     1801 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     1861 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     1921 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     1981 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     2041 attactatta ataactaatg catggcggta atacggttat ccacagaatc aggggataac
     2101 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2161 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2221 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2341 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2581 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2821 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2881 tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc
     2941 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa
     3001 aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc
     3061 cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc
     3121 tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca
     3181 gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     3241 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     3301 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     3361 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     3421 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagt tcttcagcaa
     3481 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     3541 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     3601 gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg
     3661 ctggcgcgag cccctgatgt tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     3721 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     3781 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     3841 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     3901 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     3961 atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     4021 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     4081 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     4141 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg
     4201 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
     4261 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     4321 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     4381 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     4441 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     4501 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggattct
     4561 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4621 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4681 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4741 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4801 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4861 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4921 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4981 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5041 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5101 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcgtta agatacattg
     5161 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5221 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     5281 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     5341 aaaacctcta caaatgtggt atggctgatt atgatcatga acagactgtg aggactgagg
     5401 ggcctgaaat gagccttggg actgtgaatc taaaatacac aaacaattag aatcagtagt
     5461 ttaacacatt atacacttaa aaattggatc tccattcgcc attcaggctg cgcaactgtt
     5521 gggaagggcg atcgagtgaa ttgtaatacg actc
//
