LOCUS       sequence-349791-        5236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding part of the budding yeast Rnt1 protein from L366 to S453.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5236)
  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 5236)
  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-Rnt1-dsRBD(+6A)
FEATURES             Location/Qualifiers
     source          1..5236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(146..151,152..169,170..176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    184..200
                     /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     208..224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     208..224
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             220..747
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(240..257)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(240..256)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(249..271)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(849..854,855..870,871..877)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    885..901
                     /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             924..1577
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1535..1557
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1584..1601
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1952..1974)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2119..2137)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2205..2309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2310..2378,2379..3170)
                     /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(2528..2547)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3341..3929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3830..3849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4083..4100
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4173..4250
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4251..5236,1..97)
                     /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(4270..4289)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
       61 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
      121 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      181 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca
      241 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
      301 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
      361 ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt tccggcacca
      421 gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc
      481 ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt gacctatccc
      541 attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta ctcgctcaca
      601 tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt tgatggcgtt
      661 ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa
      721 gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac
      781 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
      841 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca
      901 atttcacaca ggaaacagta ttcatgtccc ctatactagg ttattggaaa attaagggcc
      961 ttgtgcaacc cactcgactt cttttggaat atcttgaaga aaaatatgaa gagcatttgt
     1021 atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc
     1081 ccaatcttcc ttattatatt gatggtgatg ttaaattaac acagtctatg gccatcatac
     1141 gttatatagc tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt gcagagattt
     1201 caatgcttga aggagcggtt ttggatatta gatacggtgt ttcgagaatt gcatatagta
     1261 aagactttga aactctcaaa gttgattttc ttagcaagct acctgaaatg ctgaaaatgt
     1321 tcgaagatcg tttatgtcat aaaacatatt taaatggtga tcatgtaacc catcctgact
     1381 tcatgttgta tgacgctctt gatgttgttt tatacatgga cccaatgtgc ctggatgcgt
     1441 tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat cccacaaatt gataagtact
     1501 tgaaatccag caagtatata gcatggcctt tgcagggctg gcaagccacg tttggtggtg
     1561 gcgaccatcc tccaaaatcg gatctggttc cgcgtggatc ccttgatatg aatgctaaaa
     1621 ggcagcttta ctctttgatt gctgcagcgg ctgccgcagg ctatgcttca ttacgtctac
     1681 attatgttac cgtaaaaaaa cccactgcag ttgatcctaa ttccatagtt gagtgcagag
     1741 taggtgatgg aacagtttta gggaccggtg tgggcagaaa tatcaaaatt gcgggtatta
     1801 gggctgcaga aaatgctctt cgtgacaaaa aaatgttaga tttttacgcc aaacaaagag
     1861 ctgccattcc taggagtgaa tcttaactcg agcggccgca tcgtgactga ctgacgatct
     1921 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     1981 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     2041 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     2101 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     2161 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2221 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2281 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2341 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2401 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2461 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2521 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     2581 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2641 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2701 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2761 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2821 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     2881 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2941 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3001 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3061 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3121 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3181 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3241 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3301 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3361 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3421 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3481 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3541 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3601 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3661 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3721 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3781 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3841 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3901 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3961 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4021 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4081 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     4141 atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc
     4201 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     4261 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     4321 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     4381 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     4441 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     4501 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     4561 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     4621 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     4681 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     4741 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     4801 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     4861 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     4921 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     4981 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     5041 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     5101 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     5161 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     5221 tgaaaagaaa aaccac
//
