LOCUS       sequence-432620-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supression of shRNF170_1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Liu L, Neve M, Perlaza-Jimenez L, Xi X, Purcell J, Hawdon A, Conn 
            SJ, Zenker J, Tamayo P, Goodall GJ, Rosenbluh J
  TITLE     Systematic loss-of-function screens identify pathway-specific 
            functional circular RNAs.
  JOURNAL   Nat Cell Biol. 2024 Aug;26(8):1359-1372. doi: 
            10.1038/s41556-024-01467-y. Epub 2024 Aug 2.
  PUBMED    39095657
REFERENCE   2  (bases 1 to 7084)
  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: 
            "10.1038/s41556-024-01467-y"; journalName: "Nat Cell Biol"; date: 
            "2024-08"; volume: "26"; issue: "8"; pages: "1359-1372"
COMMENT     Sequence Label: shRNA_RNF170_1
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     235..255
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        237..255
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        283..509
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             510..690
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    737..862
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1355..1588
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1773..1817
                     /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"
     promoter        2115..2363
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2115..2135
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2286..2305
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2460..2577
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2626..3136
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3055..3073
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3158..3757
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3158..3177)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3654..3674
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3885..4118
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4190..4311
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4227..4246)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4281..4300
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4351..4486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4506..4525)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4507..4525)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4535..4552)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4535..4551)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4544..4566)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4693..5148
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4780..4799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4990..5011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5174..5278
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5279..5347,5348..6139)
                     /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     complement(5497..5516)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6310..6898
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6799..6818
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7052..7069
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
      241 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
      301 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
      361 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
      421 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
      481 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
      541 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      601 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      661 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      721 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      781 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      841 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
      901 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
      961 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1021 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1081 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1141 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1201 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1261 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1321 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1381 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1441 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1501 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1561 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1621 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     1681 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     1741 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     1801 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     1861 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     1921 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     1981 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     2041 gatccattcg attagtgaac ggatctcgac ggtatcgatc acgagactag cctcgagcgg
     2101 ccgccccctt caccgagggc ctatttccca tgattccttc atatttgcat atacgataca
     2161 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
     2221 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     2281 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     2341 tcttgtggaa aggacgaaac accgggagcg gctctagacc tggaatctcg agattccagg
     2401 tctagagccg ctctttttga attctcgacc tcgagacaaa tggcagtatt catccacaat
     2461 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     2521 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     2581 gtttattaca gggacagcag agatccactt tggccgcggc tcgagggggt tggggttgcg
     2641 ccttttccaa ggcagccctg ggtttgcgca gggacgcggc tgctctgggc gtggttccgg
     2701 gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc acgtccgttc gcagcgtcac
     2761 ccggatcttc gccgctaccc ttgtgggccc cccggcgacg cttcctgctc cgcccctaag
     2821 tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct
     2881 cactagtacc ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg
     2941 ggctgtggcc aatagcggct gctcagcagg gcgcgccgag agcagcggcc gggaaggggc
     3001 ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc
     3061 cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg accgaatcac
     3121 cgacctctct ccccaggggg atccaccgga gcttaccatg accgagtaca agcccacggt
     3181 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     3241 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     3301 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     3361 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     3421 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     3481 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     3541 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     3601 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     3661 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     3721 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg
     3781 cccgaccgaa aggagcgcac gaccccatgc atcggtacct ttaagaccaa tgacttacaa
     3841 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     3901 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     3961 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     4021 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     4081 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     4141 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     4201 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4261 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4321 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4381 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4441 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca
     4501 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     4561 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     4621 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     4681 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4741 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4801 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4861 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4921 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4981 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5041 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5101 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     5161 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5221 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5281 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5341 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5401 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5461 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5521 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5581 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5641 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5701 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5761 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5821 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5881 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5941 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6001 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6061 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6121 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6181 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6241 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6301 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6361 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6421 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6481 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6541 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6601 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6661 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6721 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6781 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6841 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6901 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6961 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     7021 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     7081 gcgc
//
