LOCUS       sequence-343345-        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-ts2
FEATURES             Location/Qualifiers
     source          1..6319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(59..79)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        64..393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      244..379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     306..325
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        405..784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        786..984
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     939..959
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1002..1182
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1229..1354
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1851..2084
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2269..2313
                     /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    2611..2728
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2865..3272
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     ncRNA           3987..4001
                     /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             4135..4315
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4386..4974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4466..4485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5145..5936,5937..6005))
                     /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     5768..5787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6006..6110)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6185..6204)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgctgtgg
       61 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
      301 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
      361 ggaggctttt ttggaggcct aggcttttgc aaaaagctcg cgttgacatt gattattgac
      421 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      481 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      541 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      601 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      661 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      721 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      781 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      841 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      901 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      961 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
     1021 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     1081 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgagactct ggtaactaga
     1141 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     1201 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     1261 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     1321 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     1381 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     1441 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     1501 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1561 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1621 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1681 cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat gagggacaat
     1741 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     1801 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     1861 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     1921 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     1981 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     2041 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2101 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2161 ctggaacaga tttggaatca cacgacttgg atggagtggg acagagaaat taacaattac
     2221 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2281 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2341 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2401 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2461 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     2521 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     2581 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     2641 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     2701 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     2761 ttggttaatt aactgcagcc ccgataaaat aaaagatttt atttagtctc cagaaaaagg
     2821 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc tgcagtaacg ccattttgca
     2881 aggcatggaa aaataccaaa ccaagaatag agaagttcag atcaagggcg ggtacatgaa
     2941 aatagctaac gttgggccaa acaggatatc tgcggtgagc agtttcggcc ccggcccggg
     3001 gccaagaaca gatggtcacc gcagtttcgg ccccggcccg aggccaagaa cagatggtcc
     3061 ccagatatgg cccaaccctc agcagtttct taagacccat cagatgtttc caggctcccc
     3121 caaggacctg aaatgaccct gcgccttatt tgaattaacc aatcagcctg cttctcgctt
     3181 ctgttcgcgc gcttctgctt cccgagctct ataaaagagc tcacaacccc tcactcggcg
     3241 cgccagtcct ccgacagact gagtcgcccg ggggtctaga acgcgtgccg ccatgaccga
     3301 atacaaacct accgtgaggc tggctacaag agatgatgtc ccaagggctg tgagaacact
     3361 ggccgccgct tttgccgatt accctgccac acgccacact gtggacccag atcggcatat
     3421 cgagagagtg actgagctgc aggaactgtt cctgacccga gtgggcctgg acattgggaa
     3481 ggtctgggtc gcagacgatg gagcagctgt ggctgtctgg accacaccag agagcgtgga
     3541 agccggagct gtctttgcag agatcggccc tagaatggca gaactgagcg gctccaggct
     3601 ggcagcacag cagcagatgg agggactgct ggccccacac aggcctaagg aaccagcatg
     3661 gttcctggct accgtggggg tctctcctga ccatcagggc aaaggactgg gaagtgctgt
     3721 ggtcctgcca ggagtggagg ctgcagaacg agctggagtc cctgcatttc tggagacctc
     3781 tgctccacga aacctgccct tctatgaacg gctgggcttt actgtgaccg cagatgtgga
     3841 ggtccccgaa ggacctagga cctggtgcat gacacgcaaa cccggcgcct gagcgatcgc
     3901 cgcggccgcc ttcttaaccc aacagaaggc tcgagaaggt atattgctgt tgacagtgag
     3961 cgccaataca gtcaagctgg cttatagtga agccacagat gtataagcca gcttgactgt
     4021 attgttgcct actgcctcgg acttcaaggg gctagaattc ggcagctgta gatcttagcc
     4081 actttttaaa agaaaagggg ggactggaag ggctaactgc atccggactg tactgggtct
     4141 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     4201 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     4261 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc
     4321 cgtttcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4381 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4441 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4501 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4561 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4621 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4681 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4741 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4801 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4861 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4921 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4981 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5041 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5101 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5161 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5221 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5281 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5341 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5401 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5461 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5521 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5581 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5641 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5701 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5761 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5821 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5881 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5941 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6001 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6061 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6121 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     6181 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     6241 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     6301 acaattgcat gaagaatct
//
