LOCUS       sequence-339638-        5227 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 5227)
  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 5227)
  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(+3A)
FEATURES             Location/Qualifiers
     source          1..5227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(564..569,570..587,588..594)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    602..618
                     /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     626..642
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..642
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             638..1165
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(658..675)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(658..674)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(667..689)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1267..1272,1273..1288,1289..1295)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1303..1319
                     /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             1342..1995
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1953..1975
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2002..2019
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2361..2383)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2528..2546)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2614..2718
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2719..2787,2788..3579)
                     /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(2937..2956)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3750..4338
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4239..4258
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4492..4509
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4582..4659
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4660..5227,1..515)
                     /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(4679..4698)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
       61 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
      121 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
      181 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
      241 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
      301 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
      361 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
      421 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      481 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
      541 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      601 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cggattcact
      661 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
      721 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
      781 ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga
      841 agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc
      901 ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtga cctatcccat
      961 tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt
     1021 taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttgg
     1081 aattagctta tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc
     1141 tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc
     1201 ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat
     1261 gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc ggataacaat
     1321 ttcacacagg aaacagtatt catgtcccct atactaggtt attggaaaat taagggcctt
     1381 gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat
     1441 gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc
     1501 aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt
     1561 tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca
     1621 atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa
     1681 gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc
     1741 gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc
     1801 atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc
     1861 ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg
     1921 aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc
     1981 gaccatcctc caaaatcgga tctggttccg cgtggatccc ttgatatgaa tgctaaaagg
     2041 cagctttact ctttgattgc tgcagcgggc tatgcttcat tacgtctaca ttatgttacc
     2101 gtaaaaaaac ccactgcagt tgatcctaat tccatagttg agtgcagagt aggtgatgga
     2161 acagttttag ggaccggtgt gggcagaaat atcaaaattg cgggtattag ggctgcagaa
     2221 aatgctcttc gtgacaaaaa aatgttagat ttttacgcca aacaaagagc tgccattcct
     2281 aggagtgaat cttaactcga gcggccgcat cgtgactgac tgacgatctg cctcgcgcgt
     2341 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2401 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2461 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga
     2521 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2581 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2641 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2701 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2761 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2821 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2881 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2941 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     3001 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3061 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3121 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3181 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3241 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     3301 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3361 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3421 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3481 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3541 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3601 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3661 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3721 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3781 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3841 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3901 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3961 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4021 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4081 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4141 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4201 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4261 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4321 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4381 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4441 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4501 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4561 atttcacacc gcataaattc cgacaccatc gaatggcgca aaacctttcg cggtatggca
     4621 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
     4681 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     4741 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     4801 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     4861 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     4921 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     4981 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     5041 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     5101 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     5161 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     5221 ccattaa
//
