LOCUS       sequence-339627-        5215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a Dicer fragment extending from from R1841 to the 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5215)
  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 5215)
  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-Dicer
FEATURES             Location/Qualifiers
     source          1..5215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     505..524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     758..775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        848..925
                     /gene="lacI"
                     /label=lacI promoter
     CDS             926..2008
                     /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(945..964)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2057..2062,2063..2080,2081..2087)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2095..2111
                     /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     2119..2135
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2119..2135
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2131..2658
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2151..2168)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2151..2167)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2160..2182)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2760..2765,2766..2781,2782..2788)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2796..2812
                     /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             2835..3488
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3446..3468
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3495..3512
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3842..3864)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4009..4027)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4095..4199
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4200..4268,4269..5060)
                     /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(4418..4437)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
       61 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      121 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
      181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      781 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
      841 aaattccgac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga
      901 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
      961 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     1021 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     1081 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     1141 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     1201 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     1261 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     1321 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     1381 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     1441 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     1501 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     1561 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     1621 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     1681 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     1741 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
     1801 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     1861 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     1921 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     1981 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
     2041 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2101 gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc
     2161 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2221 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2281 cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc agaagcggtg
     2341 ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac
     2401 tggcagatgc acggttacga tgcgcccatc tacaccaacg tgacctatcc cattacggtc
     2461 aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt
     2521 gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt tggaattagc
     2581 ttatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga agctgtggta
     2641 tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct
     2701 ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga aatgagctgt
     2761 tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac aatttcacac
     2821 aggaaacagt attcatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac
     2881 ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg
     2941 atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc
     3001 cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag
     3061 ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg
     3121 aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg
     3181 aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc
     3241 gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt
     3301 atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat
     3361 tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca
     3421 gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc
     3481 ctccaaaatc ggatctggtt ccgcgtggat ccccggaatt ccggccacta atagaaaagt
     3541 tttctgcaaa tgtaccccgt tcccctgtgc gagaattgct tgaaatggaa ccagaaactg
     3601 ccaaatttag cccggctgag agaacttacg acgggaaggt cagagtcact gtggaagtag
     3661 taggaaaggg gaaatttaaa ggtgttggtc gaagttacag gattgccaaa tctgcagcag
     3721 caagaagagc cctccgaagc ctcaaagcta atcaacctca ggttcccaat agctgactcg
     3781 agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa
     3841 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3901 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     3961 cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc ctcgtgatac
     4021 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4081 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4141 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4201 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4261 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4321 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4381 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4441 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4501 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4561 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4621 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4681 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4741 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4801 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4861 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4921 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4981 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5041 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5101 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5161 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaa
//
