LOCUS       sequence-177324-        6257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Flag-MCPIP1(81-599).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6257)
  AUTHORS   Liang J, Saad Y, Lei T, Wang J, Qi D, Yang Q, Kolattukudy PE, Fu M
  TITLE     MCP-induced protein 1 deubiquitinates TRAF proteins and negatively 
            regulates JNK and NF-kappaB signaling.
  JOURNAL   J Exp Med. 2010 Dec 20;207(13):2959-73. doi: 10.1084/jem.20092641. 
            Epub 2010 Nov 29.
  PUBMED    21115689
REFERENCE   2  (bases 1 to 6257)
  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: 
            "10.1084/jem.20092641"; journalName: "J Exp Med"; date: "2010-12-20-
            20"; volume: "207"; issue: "13"; pages: "2959-73"
COMMENT     Sequence Label: Flag-MCPIP1(81-599)
FEATURES             Location/Qualifiers
     source          1..6257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             645..668
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2281..2903
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2480..2499)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2927..2947)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2932..3261
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3112..3247
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3299..3318)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3300..3318)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3332..3348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3332..3348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3345..3367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3356..3372
                     /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(3380..3386,3387..3404,3405..3410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3425..3446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3563..3580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3734..4322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3814..3833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4493..5284,5285..5353))
                     /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     5116..5135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5354..5458)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5485..5940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5572..5591)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5782..5803
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6066..6088
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6080..6097
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6081..6097
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgatc taccatggcc cacaaccacc gccacaagca cgccgactac aaagacgatg
      661 acgacaagct tgcggccgcg aattcaatgc tgggtgagct ggtgaaacac gggacagcca
      721 ccgagcggga gcgccagacc tcaccggacc cctgccctca gctccctcta gtcccgcggg
      781 gtggtggcac ccctaaggct cccaacctgg agcctccact cccagaagag gaaaaggagg
      841 gcagcgacct gagaccagtg gtcatcgatg ggagcaacgt ggccatgagc catgggaaca
      901 aggaggtctt ctcctgccgg ggcatcctgc tggcagtgaa ctggtttctg gagcggggcc
      961 acacagacat cacagtgttt gtgccatcct ggaggaagga gcagcctcgg cccgacgtgc
     1021 ccatcacaga ccagcacatc ctgcgggaac tggagaagaa gaagatcctg gtgttcacac
     1081 catcacgacg cgtgggtggc aagcgggtgg tgtgctatga cgacagattc attgtgaagc
     1141 tggcctacga gtctgacggg atcgtggttt ccaacgacac ataccgtgac ctccaaggcg
     1201 agcggcagga gtggaagcgc ttcatcgagg agcggctgct catgtactcc ttcgtcaatg
     1261 acaagtttat gccccctgat gacccactgg gccggcacgg gcccagcctg gacaacttcc
     1321 tgcgtaagaa gccactcact ttggagcaca ggaagcagcc gtgtccctat ggaaggaaat
     1381 gcacctatgg gatcaagtgc cgattcttcc acccagagcg gccaagctgc ccccagcgct
     1441 ctgtggcaga tgagctccgt gccaatgctc tcctctcacc ccccagagcc ccaagcaagg
     1501 acaaaaatgg ccggcggcct tcaccttcat cccagtccag ctctctgcta acagagagtg
     1561 agcagtgcag cctggatggg aagaagctgg gggcccaggc atccccaggg tcccgccaag
     1621 agggtctaac acagacctat gccccatcag gcaggagcct cgcacctagc gggggcagtg
     1681 gcagcagctt tgggcccaca gactggctcc cacagacgct ggactcactc ccgtacgtct
     1741 cccaggattg cctggactcg ggcattggct ccctggagag ccagatgtcg gaactttggg
     1801 gggttcgagg aggaggccct ggtgagccgg gcccaccccg agccccttac acgggctaca
     1861 gtccctatgg atctgagctc ccagccaccg cagccttctc tgcctttggc cgggccatgg
     1921 gtgctggcca cttcagtgtc cctgccgact acccacccgc gccccctgcc tttccacctc
     1981 gagagtactg gtctgaacca tacccactgc ccccacccac atcagtcctt caggagcccc
     2041 cagtgcagag cccaggggct ggcaggagcc cgtggggcag ggcaggcagc ctggccaagg
     2101 agcaggccag cgtgtatact aagctgtgtg gtgtgtttcc cccgcacctg gtggaggctg
     2161 tgatggggcg cttcccacag ctcctggacc cccagcagct ggctgccgag atcctctcct
     2221 acaagtccca gcaccccagt gagtaatcta gatatcggta ccggatccag atctgatccc
     2281 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     2341 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     2401 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     2461 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     2521 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     2581 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     2641 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     2701 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     2761 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     2821 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc
     2881 actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag
     2941 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3001 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3061 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3121 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3181 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3241 ggaggcctag gcttttgcaa aaagctcctc gaggaactga aaaaccagaa agttaattcc
     3301 ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt
     3361 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3421 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3481 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3541 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3601 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3901 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4141 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4381 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4441 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4501 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4561 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4621 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4681 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4741 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4801 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4861 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4921 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4981 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5041 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5101 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5161 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5221 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5281 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5341 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5401 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5461 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5521 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5581 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5641 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5701 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5761 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5821 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5881 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5941 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6001 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6061 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
     6121 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
     6181 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
     6241 aatatgaccg ccatgtt
//
