LOCUS       sequence-432515-        7084 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Supression of shcircMTO1(2,RI,3)_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 Apr 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_circMTO1_2
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    159..284
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    777..1010
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1195..1239
                     /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        1537..1785
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1537..1557
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1708..1727
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    1882..1999
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2048..2558
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2477..2495
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2580..3179
                     /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(2580..2599)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3076..3096
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3307..3540
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3612..3733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3649..3668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3703..3722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3773..3908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3928..3947)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3929..3947)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3957..3974)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3957..3973)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3966..3988)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4115..4570
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4202..4221)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4412..4433
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4596..4700
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4701..4769,4770..5561)
                     /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(4919..4938)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5732..6320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6221..6240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6474..6491
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6608..6629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6644..6649,6650..6667,6668..6674)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6682..6698
                     /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     6687..6709
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6706..6722
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6706..6722
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6741..6761
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6743..6761
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6789..7015
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             join(7016..7084,1..112)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
       61 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      121 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      181 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      241 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      301 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      361 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      421 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      481 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      541 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      601 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      661 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
      721 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
      781 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
      841 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
      901 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
      961 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1021 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1081 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1141 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1201 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1261 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1321 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1381 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     1441 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcga
     1501 tcacgagact agcctcgagc ggccgccccc ttcaccgagg gcctatttcc catgattcct
     1561 tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta
     1621 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     1681 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     1741 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggtgt tttgggtcag
     1801 atgtcatgct cgagcatgac atctgaccca aaacattttt gaattctcga cctcgagaca
     1861 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1921 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1981 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac tttggccgcg
     2041 gctcgagggg gttggggttg cgccttttcc aaggcagccc tgggtttgcg cagggacgcg
     2101 gctgctctgg gcgtggttcc gggaaacgca gcggcgccga ccctgggtct cgcacattct
     2161 tcacgtccgt tcgcagcgtc acccggatct tcgccgctac ccttgtgggc cccccggcga
     2221 cgcttcctgc tccgccccta agtcgggaag gttccttgcg gttcgcggcg tgccggacgt
     2281 gacaaacgga agccgcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa
     2341 tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg
     2401 agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc
     2461 tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg
     2521 gctccctcgt tgaccgaatc accgacctct ctccccaggg ggatccaccg gagcttacca
     2581 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     2641 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     2701 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     2761 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     2821 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     2881 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     2941 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     3001 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     3061 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     3121 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     3181 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcggtac
     3241 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     3301 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     3361 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     3421 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     3481 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     3541 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     3601 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3661 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3721 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     3781 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3841 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3901 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     3961 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4021 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4081 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     4141 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4201 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4261 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4321 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4381 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4441 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4501 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4561 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4621 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4681 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4741 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4801 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4861 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4921 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4981 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5041 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5101 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5161 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5221 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5281 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5341 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5401 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5461 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5521 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5581 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5641 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5701 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5761 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5821 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5881 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5941 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6001 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6061 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6301 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6361 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6421 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6481 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6541 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6601 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     6661 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     6721 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
     6781 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
     6841 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
     6901 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
     6961 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
     7021 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     7081 taag
//
