LOCUS       sequence-177331-        5612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Flag-MCPIP1(150-453).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5612)
  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 5612)
  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(150-453)
FEATURES             Location/Qualifiers
     source          1..5612
                     /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    1636..2258
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1835..1854)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2282..2302)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2287..2616
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2467..2602
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2654..2673)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2655..2673)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2687..2703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2687..2703)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2700..2722)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2711..2727
                     /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(2735..2741,2742..2759,2760..2765))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2780..2801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2918..2935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3089..3677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3169..3188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3848..4639,4640..4708))
                     /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     4471..4490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4709..4813)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4840..5295
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4927..4946)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5137..5158
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5421..5443
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5435..5452
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5436..5452
                     /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 aattcaatga acaaggaggt cttctcctgc cggggcatcc
      721 tgctggcagt gaactggttt ctggagcggg gccacacaga catcacagtg tttgtgccat
      781 cctggaggaa ggagcagcct cggcccgacg tgcccatcac agaccagcac atcctgcggg
      841 aactggagaa gaagaagatc ctggtgttca caccatcacg acgcgtgggt ggcaagcggg
      901 tggtgtgcta tgacgacaga ttcattgtga agctggccta cgagtctgac gggatcgtgg
      961 tttccaacga cacataccgt gacctccaag gcgagcggca ggagtggaag cgcttcatcg
     1021 aggagcggct gctcatgtac tccttcgtca atgacaagtt tatgccccct gatgacccac
     1081 tgggccggca cgggcccagc ctggacaact tcctgcgtaa gaagccactc actttggagc
     1141 acaggaagca gccgtgtccc tatggaagga aatgcaccta tgggatcaag tgccgattct
     1201 tccacccaga gcggccaagc tgcccccagc gctctgtggc agatgagctc cgtgccaatg
     1261 ctctcctctc accccccaga gccccaagca aggacaaaaa tggccggcgg ccttcacctt
     1321 catcccagtc cagctctctg ctaacagaga gtgagcagtg cagcctggat gggaagaagc
     1381 tgggggccca ggcatcccca gggtcccgcc aagagggtct aacacagacc tatgccccat
     1441 caggcaggag cctcgcacct agcgggggca gtggcagcag ctttgggccc acagactggc
     1501 tcccacagac gctggactca ctcccgtacg tctcccagga ttgcctggac tcgggcattg
     1561 gctccctgga gagccagatg tcggaacttt ggggggttta atctagatat cggtaccgga
     1621 tccagatctg atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     1681 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     1741 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     1801 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     1861 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     1921 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     1981 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     2041 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     2101 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     2161 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     2221 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcttggctg
     2281 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2341 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2401 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2461 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2521 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     2581 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcctcgagga actgaaaaac
     2641 cagaaagtta attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc
     2701 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2761 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2821 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2881 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3781 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3841 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3901 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3961 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4021 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4141 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4261 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4321 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4501 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4681 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4741 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4801 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     4861 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4921 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4981 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5041 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5101 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5161 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5221 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5281 taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5341 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5401 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     5461 gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata
     5521 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta
     5581 tattggctca tgtccaatat gaccgccatg tt
//
