LOCUS       sequence-339626-        5821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a DGCR8 fragment extending from E502 to the C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5821)
  AUTHORS   Zhang X, Li P, Zhong H, Yang F, Liu F, Yedid G, Zeng Y
  TITLE     Extending the L1 region in canonical double-stranded RNA-binding 
            domains impairs their functions.
  JOURNAL   Acta Biochim Biophys Sin (Shanghai). 2021 Mar 26;53(4):463-471. doi:
            10.1093/abbs/gmab014.
  PUBMED    33751023
REFERENCE   2  (bases 1 to 5821)
  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"; journalName: 
            "Acta Biochim Biophys Sin (Shanghai)"; date: "2021-03-26- 26"; 
            volume: "53"; issue: "4"; pages: "463-471"
COMMENT     Sequence Label: pGEX-4T-1-DGCR8-2xdsRBD(+6)
FEATURES             Location/Qualifiers
     source          1..5821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(479..484,485..502,503..509)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    517..533
                     /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     541..557
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     541..557
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             553..1080
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(573..590)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(573..589)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(582..604)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1182..1187,1188..1203,1204..1210)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1218..1234
                     /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)."
     CDS             1257..1910
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1868..1890
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1917..1934
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2870..2892)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3037..3055)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3123..3227
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3228..3296,3297..4088)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3446..3465)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4259..4847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4748..4767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5001..5018
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5091..5168
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5169..5821,1..430)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5188..5207)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
       61 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
      121 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
      181 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
      241 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
      301 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
      361 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      421 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt
      481 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      541 caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
      601 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
      661 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
      721 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
      781 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
      841 gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc
      901 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
      961 gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc
     1021 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     1081 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     1141 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc
     1201 tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt
     1261 cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg
     1321 aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc
     1381 gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg
     1441 atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt
     1501 tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata
     1561 ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt
     1621 ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat
     1681 atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg
     1741 ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac
     1801 gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc
     1861 ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg
     1921 ttccgcgtgg atccccggaa ttcgagtttg ttattaaccc caacgggaaa tccgaggtct
     1981 gcatcctgca cgagtacatg cagcgtgtcc tcaagaccct tcggactgaa ggtgtccgcc
     2041 ctgtctataa tttctttgaa tgtgagaacc caagtgagcc ttttggtgcc tcggtgacca
     2101 ttgatggtgt gacttacgga tctggaactg caagcagcaa aaaacttgcg aagaataaag
     2161 ctgcccgagc tacactggaa atcctcatcc ctgactttgt taaacagacc tctgaagaga
     2221 agcccaaaga cagtgaagaa ctcgagtatt ttaaccacat cagcatcgag gactcgcggg
     2281 tctacgagct gaccagcaag gctgggctgt tgtctccata tcagatcctc cacgagtgcc
     2341 ttaaaagaaa ccatgggatg acccttcgga ctgaaggtgg tgacacgtct atcaagtttg
     2401 aagtggttcc tgggaaaaac cagaagagtg aatacgtcat ggcgtgtggc aagcacacag
     2461 tgcgcgggtg gtgtaagaac aagagagttg gaaagcagtt agcctcacag aagatccttc
     2521 agctgctgca cccacatgtc aagaactggg ggtctttact gcgcatgtat ggccgtgaga
     2581 gcagcaagat ggtcaagcag gagacatcgg acaagagtgt gattgagctg cagcagtatg
     2641 ccaagaagaa caagcccaac ctgcacatcc tcagcaagct ccaagaggag atgaagaggc
     2701 tagctgagga aagggaggag actcgaaaga agcccaagat gtccattgtg gcgtccgccc
     2761 agcctggcgg tgagcccctg tgcaccgtgg acgtgtgagt taacgtcgag cggccgcatc
     2821 gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2881 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2941 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     3001 gcgatagcgg agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     3061 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3121 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3181 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3241 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3301 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3361 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3421 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     3481 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3541 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3601 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3661 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3721 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     3781 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3841 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3901 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3961 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4021 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4081 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4141 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4201 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4261 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4321 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4381 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4441 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4501 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4561 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4621 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4681 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4741 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4801 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4861 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4921 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4981 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     5041 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataaattcc gacaccatcg
     5101 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
     5161 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
     5221 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
     5281 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
     5341 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     5401 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     5461 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     5521 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     5581 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     5641 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     5701 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     5761 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     5821 g
//
