LOCUS       sequence-191031-        6157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to express MIP-T3 in mammalian system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6157)
  AUTHORS   Ng MH, Ho TH, Kok KH, Siu KL, Li J, Jin DY
  TITLE     MIP-T3 is a negative regulator of innate type I IFN response.
  JOURNAL   J Immunol. 2011 Dec 15;187(12):6473-82. doi: 
            10.4049/jimmunol.1100719. Epub 2011 Nov 11.
  PUBMED    22079989
REFERENCE   2  (bases 1 to 6157)
  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.1100719"; journalName: "J Immunol"; date: 
            "2011-12-15- 15"; volume: "187"; issue: "12"; pages: "6473-82"
COMMENT     Sequence Label: MIP-T3
FEATURES             Location/Qualifiers
     source          1..6157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     495..514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        769..1072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1073..1276
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1227..1247
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1322..1340
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      1375..1384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1384..1407
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3360..3379)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3361..3379)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3653..3774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3690..3709)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3744..3763
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3781..4236)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3918..3939)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4130..4149
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4263..4367
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4369..4726
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4369..4560
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4392..4412)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4577..4712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4761..5555
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4761..5555
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4815..4834)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5425..5444
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5743..5762)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5787..5834
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
       61 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      661 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
      721 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      781 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      841 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      901 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      961 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1021 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1081 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1141 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     1201 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     1261 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgattac gccaagctcg
     1321 aaattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgccacc
     1381 atggattaca aggatgacga cgataagagc ccgggcggat ccatgaacgc ggcggtggtg
     1441 aggcggacgc aggaggcgct ggggaaagtg attcggaggc cgccgctgac agagaagctg
     1501 ctgagcaagc ccccgttccg ctacctgcac gacatcatca cggaggtgat tagaatgact
     1561 ggtttcatga agggcctcta cacagacgcc gagatgaagt ctgataatgt gaaggataaa
     1621 gatgcaaaaa ttagcttcct acaaaaggcc atagacgtgg ttgtaatggt gtcgggagag
     1681 ccactgttgg ccaaaccagc ccgaatcgtg gcggggcatg agcctgaaag aacaaacgag
     1741 ctgctccaga taattggaaa atgctgtctc aacaagctct ctagtgacga tgcggtgcgg
     1801 agggttttag ctggagagaa gggagaagtg aaaggccggg cctcactgac ctcaagatct
     1861 caggaattgg ataataagaa tgtgcgagaa gaagagtcca gagttcacaa aaatacagag
     1921 gatagaggag acgctgaaat aaaagagaga agtacaagca gagatcgaaa acagaaggaa
     1981 gaattgaaag aagaccgcaa gccaagagaa aaggacaagg acaaggagaa ggccaaggag
     2041 aatggcggaa acagacacag agaaggggag agagagagag ccaaagcccg ggccaggcca
     2101 gacagcgagc gacagaaaga cagaggcaac agggagcggg acagagactc cgagcgcaag
     2161 aaggagacag agagaaagag tgaggggggg aaagagaagg agagactgag agacagggac
     2221 cgagagcgcg accgggacaa agggaaggac agggacagac ggagagtgaa aaacggggag
     2281 cactcctggg acctggacag ggagaagaac agagagcatg acaaacccga gaaaaagtca
     2341 gcaagctcag gggagatgtc taaaaagtta tcagatggaa cttttaaaga ctccaaggct
     2401 gaaacagaga ctgagatttc cactagagct tccaagtcat tgacaacaaa aacatcaaaa
     2461 cggcgatcca aaaattcagt ggaaggtgac tccaccagtg atgcagaagg agatgctgga
     2521 cctgctggcc aagataagtc tgaggtgcca gagactccag aaattcctaa tgagctttca
     2581 tccaacatca gaagaattcc tcggcctggg agtgcaagac cagcccctcc ccgggtcaaa
     2641 cggcaagaca gcatggaggc gctacaaatg gataggtcag ggagtggtaa aaccgtttca
     2701 aatgtgatta cagagtcaca caattctgac aatgaagagg atgatcaatt tgtggtggaa
     2761 gctgcccctc agctctctga aatgtcagaa attgaaatgg taacagcagt ggaactagaa
     2821 gaagaggaga agcatggtgg acttgtgaaa aaaattttgg agacgaagaa agattatgag
     2881 aaattgcagc agtcacccaa acctggggag aaggagcgat ctctctttga gtcggcatgg
     2941 aagaaggaga aggacatcgt ttccaaggag atagagaagc tccgcacgtc catccagacc
     3001 ctgtgcaaga gcgcacttcc cctggggaag atcatggact acatccagga agacgtggat
     3061 gccatgcaga atgagctgca gatgtggcac agcgagaaca ggcagcacgc cgaggccctg
     3121 cagcaggagc agaggatcac agactgtgcc gtggagccct taaaggctga gctcgcggag
     3181 ctggagcagc tgatcaaaga ccagcaagac aagatctgtg ctgtgaaggc caacatcctc
     3241 aagaatgaag aaaaaatcca gaaaatggta tatagtatca atttgacttc gagaaggtga
     3301 ctcgaggggg ggcccggtac cttaattaat taaggtacca ggtaagtgta cccaattcgc
     3361 cctatagtga gtcgtattac aattcactcg atcgcccttc ccaacagttg cgcagcctga
     3421 atggcgaatg gagatccaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt
     3481 ttgtgtattt tagattcaca gtcccaaggc tcatttcagg cccctcagtc ctcacagtct
     3541 gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3601 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3661 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3721 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     3781 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3841 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3901 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3961 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4021 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4081 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4141 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4201 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4261 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4321 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagaatcct gaggcggaaa
     4381 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4441 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4501 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4561 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4621 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4681 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4741 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4801 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4861 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4921 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4981 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5041 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5101 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5161 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5221 atcaggatga tctggacgaa gaacatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5281 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5341 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5401 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gaacttggcg
     5461 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5521 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5581 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5641 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5701 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5761 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5821 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5881 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5941 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6001 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6061 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6121 ccactgagcg tcagaccccg tagaaaagat caaagga
//
