LOCUS       sequence-343753-        6319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TMEM173 knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6319)
  AUTHORS   Guney MH, Nagalekshmi K, McCauley SM, Carbone C, Aydemir O, Luban J
  TITLE     IFIH1 (MDA5) is required for innate immune detection of 
            intron-containing RNA expressed from the HIV-1 provirus.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Jul 16;121(29):e2404349121. doi: 
            10.1073/pnas.2404349121. Epub 2024 Jul 10.
  PUBMED    38985764
REFERENCE   2  (bases 1 to 6319)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2024-07-16"; volume: "121(29):e2404349121. 
            doi"; pages: " 10.1073/pnas.2404349121. Epub 2024 Jul 10"
COMMENT     Sequence Label: pAPM-miR30-TMEM173-ts1
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..66
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        146..525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        527..725
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             743..923
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    970..1095
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1592..1825
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2010..2054
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2352..2469
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2606..3013
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3728..3742
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     LTR             3876..4056
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4127..4715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4207..4226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4886..5677,5678..5746))
                     /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     5509..5528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5747..5851)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5926..5945)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6119..6139)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6124..6319,1..134)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6304..6319,1..120)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
       61 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      121 taggcttttg caaaaagctc gcgttgacat tgattattga ctagttatta atagtaatca
      181 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      241 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      301 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      361 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      421 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      481 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      541 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      601 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      661 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
      781 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
      841 agtagtgtgt gcccgtctgt tgtgagactc tggtaactag agatccctca gaccctttta
      901 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
      961 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     1021 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     1081 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     1141 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     1201 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     1261 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     1321 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     1381 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccggc
     1441 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1501 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1561 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1621 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1681 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1741 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1801 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     1861 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     1921 acacgacttg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1981 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     2041 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     2101 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     2161 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2221 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2281 ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta
     2341 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     2401 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     2461 caaaattttc gggtttatta cagggacagc agagatccag tttggttaat taactgcagc
     2521 cccgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     2581 ctgtaggttt ggcaagctag ctgcagtaac gccattttgc aaggcatgga aaaataccaa
     2641 accaagaata gagaagttca gatcaagggc gggtacatga aaatagctaa cgttgggcca
     2701 aacaggatat ctgcggtgag cagtttcggc cccggcccgg ggccaagaac agatggtcac
     2761 cgcagtttcg gccccggccc gaggccaaga acagatggtc cccagatatg gcccaaccct
     2821 cagcagtttc ttaagaccca tcagatgttt ccaggctccc ccaaggacct gaaatgaccc
     2881 tgcgccttat ttgaattaac caatcagcct gcttctcgct tctgttcgcg cgcttctgct
     2941 tcccgagctc tataaaagag ctcacaaccc ctcactcggc gcgccagtcc tccgacagac
     3001 tgagtcgccc gggggtctag aacgcgtgcc gccatgaccg aatacaaacc taccgtgagg
     3061 ctggctacaa gagatgatgt cccaagggct gtgagaacac tggccgccgc ttttgccgat
     3121 taccctgcca cacgccacac tgtggaccca gatcggcata tcgagagagt gactgagctg
     3181 caggaactgt tcctgacccg agtgggcctg gacattggga aggtctgggt cgcagacgat
     3241 ggagcagctg tggctgtctg gaccacacca gagagcgtgg aagccggagc tgtctttgca
     3301 gagatcggcc ctagaatggc agaactgagc ggctccaggc tggcagcaca gcagcagatg
     3361 gagggactgc tggccccaca caggcctaag gaaccagcat ggttcctggc taccgtgggg
     3421 gtctctcctg accatcaggg caaaggactg ggaagtgctg tggtcctgcc aggagtggag
     3481 gctgcagaac gagctggagt ccctgcattt ctggagacct ctgctccacg aaacctgccc
     3541 ttctatgaac ggctgggctt tactgtgacc gcagatgtgg aggtccccga aggacctagg
     3601 acctggtgca tgacacgcaa acccggcgcc tgagcgatcg ccgcggccgc cttcttaacc
     3661 caacagaagg ctcgagaagg tatattgctg ttgacagtga gcgacacccc cttgcagact
     3721 ttgtatagtg aagccacaga tgtatacaaa gtctgcaagg gggtggtgcc tactgcctcg
     3781 gacttcaagg ggctagaatt cggcagctgt agatcttagc cactttttaa aagaaaaggg
     3841 gggactggaa gggctaactg catccggact gtactgggtc tctctggtta gaccagatct
     3901 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     3961 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     4021 tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttcatg tgagcaaaag
     4081 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4141 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4201 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4261 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4321 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4381 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4441 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4501 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4561 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4621 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4681 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4741 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4801 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4861 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4921 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4981 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5041 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5101 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5161 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5221 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5281 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5341 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5401 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5461 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5521 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5581 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5641 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5701 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5761 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5821 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5881 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     5941 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     6001 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     6061 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgctgtgg
     6121 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6181 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     6241 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6301 cccatcccgc ccctaactc
//
