LOCUS       sequence-129744-        7477 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 7477)
  AUTHORS   Liang Q, Deng H, Li X, Wu X, Tang Q, Chang TH, Peng H, Rauscher FJ 
            3rd, Ozato K, Zhu F
  TITLE     Tripartite motif-containing protein 28 is a small ubiquitin-related 
            modifier E3 ligase and negative regulator of IFN regulatory factor 
            7.
  JOURNAL   J Immunol. 2011 Nov 1;187(9):4754-63. doi: 10.4049/jimmunol.1101704.
            Epub 2011 Sep 21.
  PUBMED    21940674
REFERENCE   2  (bases 1 to 7477)
  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: 
            "10.4049/jimmunol.1101704"; journalName: "J Immunol"; date: 
            "2011-11-1- 1"; volume: "187"; issue: "9"; pages: "4754-63"
COMMENT     Sequence Label: pGEX-5X-TRIM28
FEATURES             Location/Qualifiers
     source          1..7477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             921..932
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3527..3549)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3694..3712)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3780..3884
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3885..3953,3954..4745)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4103..4122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4916..5504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5405..5424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5658..5675
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5748..5825
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5826..6908
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5845..5864)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6957..6962,6963..6980,6981..6987)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6995..7011
                     /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)."
     primer_bind     7019..7035
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7019..7035
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7031..7477,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(7051..7068)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7051..7067)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7060..7082)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg atcgaaggtc gtggatctat ggcggcctcc gcggcggcag
      961 cctcggcagc agcggcctcg gccgcctctg gcagcccggg cccgggcgag ggctccgctg
     1021 gcggcgaaaa gcgctccacc gccccttcgg ccgcagcctc ggcctctgcc tcagccgcgg
     1081 cgtcgtcgcc cgcggggggc ggcgccgagg cgctggagct gctggagcac tgcggcgtgt
     1141 gcagagagcg cctgcgaccc gagagggagc cccgcctgct gccctgtttg cactcggcct
     1201 gtagtgcctg cttagggccc gcggcccccg ccgccgccaa cagctcgggg gacggcgggg
     1261 cggcgggcga cggcaccgtg gtggactgtc ccgtgtgcaa gcaacagtgc ttctccaaag
     1321 acatcgtgga gaattatttc atgcgtgata gtggcagcaa ggctgccacc gacgcccagg
     1381 atgcgaacca gtgctgcact agctgtgagg ataatgcccc agccaccagc tactgtgtgg
     1441 agtgctcgga gcctctgtgt gagacctgtg tagaggcgca ccagcgggtg aagtacacca
     1501 aggaccatac tgtgcgctct actgggccag ccaagtctcg ggatggtgaa cgtactgtct
     1561 attgcaacgt acacaagcat gaaccccttg tgctgttttg tgagagctgt gatactctca
     1621 cctgccgaga ctgccagctc aatgcccaca aggaccacca gtaccagttc ttagaggatg
     1681 cagtgaggaa ccagcgcaag ctcctggcct cactggtgaa gcgccttggg gacaaacatg
     1741 caacattgca gaagagcacc aaggaggttc gcagctcaat ccgccaggtg tctgacgtac
     1801 agaagcgtgt gcaagtggat gtcaagatgg ccatcctgca gatcatgaag gagctgaata
     1861 agcggggccg tgtgctggtc aatgatgccc agaaggtgac tgaggggcag caggagcgcc
     1921 tggagcggca gcactggacc atgaccaaga tccagaagca ccaggagcac attctgcgct
     1981 ttgcctcttg ggctctggag agtgacaaca acacagccct tttgctttct aagaagttga
     2041 tctacttcca gctgcaccgg gccctcaaga tgattgtgga tcccgtggag ccacatggcg
     2101 agatgaaatt tcagtgggac ctcaatgcct ggaccaagag tgccgaggcc tttggcaaga
     2161 ttgtggcaga gcgtcctggc actaactcaa caggccctgc acccatggcc cctccaagag
     2221 ccccagggcc cctgagcaag cagggctctg gcagcagcca gcccatggag gtgcaggaag
     2281 gctatggctt tgggtcagga gatgatccct actcaagtgc agagccccat gtgtcaggtg
     2341 tgaaacggtc ccgctcaggt gagggcgagg tgagcggcct tatgcgcaag gtgccacgag
     2401 tgagccttga acgcctggac ctggacctca cagctgacag ccagccaccc gtcttcaagg
     2461 tcttcccagg cagtaccact gaggactaca accttattgt tattgaacgt ggcgctgccg
     2521 ctgcagctac cggccagcca gggactgcgc ctgcaggaac ccctggtgcc ccacccctgg
     2581 ctggcatggc cattgtcaag gaggaggaga cggaggctgc cattggagcc cctcctactg
     2641 ccactgaggg ccctgagacc aaacctgtgc ttatggctct tgcggagggt cctggtgctg
     2701 agggtccccg cctggcctca cctagtggca gcaccagctc agggctggag gtggtggctc
     2761 ctgagggtac ctcagcccca ggtggtggcc cgggaaccct ggatgacagt gccaccattt
     2821 gccgtgtctg ccagaagcca ggcgatctgg ttatgtgcaa ccagtgtgag ttttgtttcc
     2881 acctggactg tcacctgccg gccctgcagg atgtaccagg ggaggagtgg agctgctcac
     2941 tctgccatgt gctccctgac ctgaaggagg aggatggcag cctcagcctg gatggtgcag
     3001 acagcactgg cgtggtggcc aagctctcac cagccaacca gcggaaatgt gagcgtgtac
     3061 tgctggccct attctgtcac gaaccctgcc gccccctgca tcagctggct accgactcca
     3121 ccttctccct ggaccagccc ggtggcaccc tggatctgac cctgatccgt gcccgcctcc
     3181 aggagaagtt gtcacctccc tacagctccc cacaggagtt tgcccaggat gtgggccgca
     3241 tgttcaagca attcaacaag ttaactgagg acaaggcaga cgtgcagtcc atcatcggcc
     3301 tgcagcgctt cttcgagacg cgcatgaacg aggccttcgg tgacaccaag ttctctgctg
     3361 tgctggtgga gcccccgccg atgagcctgc ctggtgctgg cctgagttcc caggagctgt
     3421 ctggtggccc tggtgatggc ccctgagaat tcccgggtcg actcgagcgg ccgcatcgtg
     3481 actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3541 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3601 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     3661 atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     3721 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3781 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3841 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3901 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3961 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4021 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4081 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     4141 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4201 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4261 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4321 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4381 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     4441 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4501 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4561 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4621 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4681 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4741 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4801 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4861 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4921 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4981 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5041 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5101 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5161 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5221 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5281 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5341 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5401 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5461 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5521 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5581 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5641 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     5701 attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat
     5761 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     5821 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     5881 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     5941 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     6001 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     6061 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     6121 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     6181 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     6241 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     6301 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     6361 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     6421 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     6481 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     6541 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     6601 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     6661 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     6721 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     6781 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     6841 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6901 ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc ccaggcttta
     6961 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7021 ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg
     7081 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     7141 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     7201 cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg
     7261 cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga
     7321 tgcgcccatc tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac
     7381 ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga
     7441 aggccagacg cgaattattt ttgatggcgt tggaatt
//
