LOCUS       sequence-343217-        8455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpresion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8455)
  AUTHORS   Li Y, Yang JY, Xie X, Jie Z, Zhang L, Shi J, Lin D, Gu M, Zhou X, Li
            HS, Watowich SS, Jain A, Yun Jung S, Qin J, Cheng X, Sun SC
  TITLE     Preventing abnormal NF-kappaB activation and autoimmunity by 
            Otub1-mediated p100 stabilization.
  JOURNAL   Cell Res. 2019 Jun;29(6):474-485. doi: 10.1038/s41422-019-0174-3. 
            Epub 2019 May 13.
  PUBMED    31086255
REFERENCE   2  (bases 1 to 8455)
  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: 
            "10.1038/s41422-019-0174-3"; journalName: "Cell Res"; date: 
            "2019-06"; volume: "29"; issue: "6"; pages: "474-485"
COMMENT     Sequence Label: pCLXSN(GFP)-myc-BioID-C
FEATURES             Location/Qualifiers
     source          1..8455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     537..557
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             587..762
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    825..1024
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1225..1641
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1571..1593
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1609..1625
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1649..1658
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1658..1687
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(1691..2038,2039..2041,2042..2650)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(2719..2739)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2724..3053
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2904..3039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(3112..3114,3115..3117,3118..3831)
                     /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(3112..3114,3115..3117,3118..3828)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3157..3178)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3418..3437)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3765..3786
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3937..4530
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     intron          4875..4940
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5070..5090
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5362..5496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5399..5418)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5453..5472
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5954..6088
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6153..6170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6324..6912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6404..6423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7083..7874,7875..7943))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7706..7725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7944..8048)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8116..8134
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     8294..8313
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      421 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      481 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      541 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcaat aaaagagccc
      601 acaacccctc actcggcgcg ccagtcttcc gatagactgc gtcgcccggg tacccgtatt
      661 cccaataaag cctcttgctg tttgcatccg aatcgtggtc tcgctgttcc ttgggagggt
      721 ctcctctgag tgattgacta cccacgacgg gggtctttca tttgggggct cgtccgggat
      781 ttggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg gccagcaact
      841 tatctgtgtc tgtccgattg tctagtgtct atgtttgatg ttatgcgcct gcgtctgtac
      901 tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca
      961 cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc
     1021 tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac
     1081 gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc
     1141 cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg
     1201 tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta
     1261 ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga
     1321 cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac
     1381 ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc acctggcccg
     1441 catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc
     1501 cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc
     1561 ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc
     1621 ctcactcctt ctctaggcgc cggaattggc caccatggaa caaaaactca tctcagaaga
     1681 ggatctcgac aaggacaaca ccgtgcccct gaagctgatc gccctgctgg ccaacggcga
     1741 gttccactct ggcgagcagc tgggagagac cctgggaatg agcagagccg ccatcaacaa
     1801 gcacatccag acactgagag actggggagt ggacgtgttc accgtgcctg gcaagggcta
     1861 cagcctgcct gagcctatcc agctgctgaa cgccaagcag atcctgggac agctggatgg
     1921 cggaagcgtg gccgtgctgc ctgtgatcga ctccaccaat cagtacctgc tggacagaat
     1981 cggagagctg aagtccggcg acgcctgcat cgccgagtac cagcaggctg gcagaggagg
     2041 cagaggacgg aagtggttca gcccattcgg agccaacctg tacctgtcca tgttctggag
     2101 actggagcag ggacctgctg ctgccatcgg actgagtctg gtgatcggaa tcgtgatggc
     2161 cgaggtgctg agaaagctgg gagccgacaa ggtgagagtg aagtggccta atgacctgta
     2221 cctccaggac cgcaagctgg ctggcatcct ggtggagctg acaggcaaga caggcgatgc
     2281 cgctcagatc gtgatcggag ccggaatcaa catggccatg agaagagtgg aggagagcgt
     2341 ggtgaaccag ggctggatca ccctgcagga ggctggcatc aacctggacc ggaacaccct
     2401 ggccgccatg ctgatcagag agctgagagc cgctctggag ctgttcgagc aggagggact
     2461 ggctccttac ctgagcagat gggagaagct ggacaacttc atcaacagac ctgtgaagct
     2521 gatcatcggc gacaaggaaa tcttcggcat ctccagagga atcgacaagc agggagctct
     2581 gctgctggag caggacggaa tcatcaagcc ctggatgggc ggagaaatct ccctgagaag
     2641 cgcagagaag ctcgagcggc cgccactgtg ctggatatct gcagaattcc accacactgg
     2701 actagtggat ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2761 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2821 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2881 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2941 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3001 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
     3061 ggctgcaggt cgaggcggat ctgatcaaga gacaggatga gcatcgatac catggtgagc
     3121 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     3181 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     3241 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3301 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3361 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3421 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3481 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3541 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     3601 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     3661 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3721 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3781 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agatgcatcg
     3841 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgataaa
     3901 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     3961 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     4021 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     4081 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     4141 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     4201 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     4261 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     4321 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     4381 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     4441 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     4501 ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct
     4561 gcccagggac caccgaccca ccaccgggag gtaagctggc tgcctcgcgc gtttcggtga
     4621 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4681 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4741 cgcagccatg acccagtcac gtagcgatag cggagtgtat ctagctaggt agctagagga
     4801 tctttgtgaa ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga
     4861 tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctactgattc
     4921 taattgtttg tgtattttag attccaacct atggaactga tgaatgggag cagtggtgga
     4981 atgcctttaa tgaggaaaac ctgttttgct cagaagaaat gccatctagt gatgatgagg
     5041 ctactgctga ctctcaacat tctactcctc caaaaaagaa gagaaaggta gaagacccca
     5101 aggactttcc ttcagaattg ctaagttttt tgagtcatgc tgtgtttagt aatagaactc
     5161 ttgcttgctt tgctatttac accacaaagg aaaaagctgc actgctatac aagaaaatta
     5221 tggaaaaata tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt
     5281 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     5341 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     5401 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5461 tccaaactca tcaatgtatc ttatcatgtc tggatcagct tcagaagatg gcggagggcc
     5521 tccaacacag taattttcct cccgactctt aaaatagaaa atgtcaagtc agttaagcag
     5581 gaagtgacta actgaccgca gctggccgtg cgacatcctc ttttaattag ttgctaggca
     5641 actgccctcc agagggcagt gtggttttgc aagaggaagc aaaaagcctc tccacccagg
     5701 cctagaatgt ttccacccaa tcattactat gacaacagct gtttttttta gtattaagca
     5761 gaggccgggg gcccctggcc tccgcttact ctggagaaaa agaagagagg cattgtagag
     5821 gcttccagag gcaacttgtc aaaacaggac tgcttctatt tctgtcacac tgtctggccc
     5881 tgtcacaagg tccagcacct ccataccccc tttaataagc agtttgggaa cgggtgcggg
     5941 tcttactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6001 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6061 gaagtagtga ggaggctttt ttggaggctg cattaatgaa tcggccaacg cgcggggaga
     6121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6721 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7381 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8041 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8101 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     8161 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     8221 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     8281 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     8341 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     8401 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcgcg
//
