LOCUS       sequence-132733-        8691 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 8691)
  AUTHORS   Reiley W, Zhang M, Wu X, Granger E, Sun SC
  TITLE     Regulation of the deubiquitinating enzyme CYLD by IkappaB kinase 
            gamma-dependent phosphorylation.
  JOURNAL   Mol Cell Biol. 2005 May . 25(10):3886-95.
  PUBMED    15870263
REFERENCE   2  (bases 1 to 8691)
  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: "Mol Cell 
            Biol. 2005 May . 25(10):3886-95."
COMMENT     Sequence Label: pCLXSN-GFP
FEATURES             Location/Qualifiers
     source          1..8691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             589..764
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    827..1026
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1227..1643
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1573..1595
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1611..1627
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1693..1702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1699..1701,1702..1704,1705..2418)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1699..1701,1702..1704,1705..2415)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1744..1765)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2005..2024)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2352..2373
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2557..2577)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2562..2891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2742..2877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2951..3745
                     /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(3005..3024)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3615..3634
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             3805..4398
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     intron          4745..4810
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4939..4959
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5231..5352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5268..5287)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5322..5341
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6397..6414)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6568..7156)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6648..6667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7625..8001)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     7948..7967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8186..8290)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8358..8376
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     8536..8555
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga ctatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatgtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctca ataaaagagc
      601 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta
      661 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg
      721 gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg
      781 atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      841 cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt
      901 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      961 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
     1021 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     1081 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     1141 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     1201 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1261 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1321 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1381 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1441 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1501 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1561 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1621 ccctcactcc ttctctaggc gccggaattc tgcagtcgac ggtaccgcgg gcccgggatc
     1681 caccggccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1741 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     1801 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1861 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1921 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1981 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2041 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2101 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2161 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2221 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2281 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2341 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2401 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     2461 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2521 tgaatgcaat tgttgttgtt tcgaggatcc ggctgtggaa tgtgtgtcag ttagggtgtg
     2581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2641 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2701 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2761 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2821 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2881 gcttttgcaa aaagcttggg ctgcaggtcg aggcggatct gatcaagaga caggatgagg
     2941 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3001 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3061 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3121 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3181 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3241 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3301 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3361 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3421 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     3481 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3541 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3601 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3661 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3721 tcgccttctt gacgagttct tctgagcggg actctggggt tcgataaaat aaaagatttt
     3781 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     3841 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag
     3901 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     3961 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     4021 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     4081 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     4141 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     4201 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     4261 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     4321 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     4381 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca
     4441 ccgacccacc accgggaggt aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa
     4501 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4561 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     4621 ccagtcacgt agcgatagcg gagtgtatac ctagctaggt agctagagga tctttgtgaa
     4681 ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga tttaaagctc
     4741 taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctactgattc taattgtttg
     4801 tgtattttag attccaacct atggaactga tgaatgggag cagtggtcca atgcctttaa
     4861 tgaggaaaac ctgttttgct cagaagaaat gcctctagtg atgatgaggc tactgctgac
     4921 tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct
     4981 tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt
     5041 gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat
     5101 ttgatgtata gtgccttgac tagagatcat aatcagccat accacatttg tagaggtttt
     5161 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     5221 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5281 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5341 caatgtatct tatcatgtct gatcccgcca tggtatcaac gccatatttc tatttacagt
     5401 agggacctct tcgttgtgta ggtaccgctg tattcctagg gaaatagtag aggcaccttg
     5461 aactgtctgc atcagccata tagcccccgc tgttcgactt acaaacacag gcacagtact
     5521 gacaaaccca tacacctcct ctgaaatacc catagttgct agggctgtct ccgaactcat
     5581 tacaccctcc aaagtcagag ctgtaatttc gccatcaagg gcagcgaggg cttctccaga
     5641 taaaatagct tctgccgaga gtcccgtaag ggtagacact tcagctaatc cctcgatgag
     5701 gtctactaga atagtcagtg cggctcccat tttgaaaatt cacttacttg atcagcttca
     5761 gaagatggcg gagggcctcc aacacagtaa ttttcctccc gactcttaaa atagaaaatg
     5821 tcaagtcagt taagcaggaa gtggactaac tgacgcagct ggccgtgcga catcctcttt
     5881 taattagttg ctaggcaacg ccctccagag ggcgtgtggt tttgcaagag gaagcaaaag
     5941 cctctccacc caggcctaga atgtttccac ccaatcatta ctatgacaac agctgttttt
     6001 tttagtatta agcagaggcc ggggacccct ggcccgctta ctctggagaa aaaaaacatt
     6061 gtagaggctt ccagaggcaa cttgtcaaaa caggactgct tctatttctg tcacactgtc
     6121 tggccctgtc acaaggtcca gcacctccat acccccttta ataagcagtt tgggaacggg
     6181 tgcgggtctt actccgccca tccgccccta actccgccca gttccgccca ttctccgccc
     6241 catgctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6301 tccagaagta gtgaggaggc ttttttggag gctgcattaa tgaatcggcc aacgcgcggg
     6361 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6421 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6481 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6541 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6601 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6661 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6721 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6781 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6841 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6901 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6961 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7021 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7081 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7141 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7201 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7261 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7321 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7381 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7441 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7501 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7561 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7621 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7681 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7741 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7801 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7861 cttttctgtg actggtgaac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7921 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7981 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8041 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8101 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8161 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8221 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8281 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     8341 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     8401 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     8461 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     8521 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     8581 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     8641 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc c
//
