LOCUS       sequence-432622-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Supression of shRERE_2 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_RERE_2
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    100..121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(136..141,142..159,160..166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    174..190
                     /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     179..201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     198..214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     198..214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     233..253
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        235..253
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        281..507
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             508..688
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    735..860
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1353..1586
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1771..1815
                     /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        2113..2361
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2113..2133
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2284..2303
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2458..2575
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2624..3134
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3053..3071
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3156..3755
                     /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(3156..3175)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3652..3672
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3883..4116
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4188..4309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4225..4244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4279..4298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4349..4484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4504..4523)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4505..4523)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4533..4550)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4533..4549)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4542..4564)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4691..5146
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4778..4797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4988..5009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5172..5276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5277..5345,5346..6137)
                     /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(5495..5514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6308..6896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6797..6816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7050..7067
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
       61 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      121 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      181 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
      241 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
      301 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
      361 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
      421 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
      481 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
      541 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      601 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      661 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
      721 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
      781 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
      841 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
      901 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
      961 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1021 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1081 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1141 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1201 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     1261 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     1321 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     1381 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     1441 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     1501 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     1561 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     1621 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     1681 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     1741 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     1801 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     1861 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     1921 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     1981 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     2041 tccattcgat tagtgaacgg atctcgacgg tatcgatcac gagactagcc tcgagcggcc
     2101 gcccccttca ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     2161 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
     2221 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     2281 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     2341 ttgtggaaag gacgaaacac cggctgagac aaggaggctg aacactcgag tgttcagcct
     2401 ccttgtctca gtttttgaat tctcgacctc gagacaaatg gcagtattca tccacaattt
     2461 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     2521 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     2581 ttattacagg gacagcagag atccactttg gccgcggctc gagggggttg gggttgcgcc
     2641 ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga
     2701 aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc
     2761 ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc
     2821 gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca
     2881 ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg
     2941 ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg
     3001 tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg
     3061 cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg
     3121 acctctctcc ccagggggat ccaccggagc ttaccatgac cgagtacaag cccacggtgc
     3181 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     3241 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     3301 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     3361 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     3421 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     3481 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     3541 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     3601 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     3661 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     3721 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     3781 cgaccgaaag gagcgcacga ccccatgcat cggtaccttt aagaccaatg acttacaagg
     3841 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     3901 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     3961 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     4021 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     4081 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     4141 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     4201 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4261 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     4321 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4381 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4441 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     4501 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     4561 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     4621 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4681 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4741 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4801 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4861 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4921 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4981 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5041 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5101 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     5161 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5221 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5281 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5341 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5401 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5461 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5521 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5581 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5641 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5701 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5761 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5821 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5881 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5941 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6001 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6061 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6121 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6181 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6241 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6301 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6361 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6421 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6481 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6541 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6601 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6661 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6721 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6781 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6841 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6901 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6961 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7021 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     7081 gccc
//
