LOCUS       sequence-211459-        7183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET-pro-siRNA-V2 based plasmid for production of recombinant siRNAs 
            (pro-siRNAs) against human TP53 gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7183)
  AUTHORS   Kaur G, Cheung HC, Xu W, Wong JV, Chan FF, Li Y, McReynolds L, Huang
            L
  TITLE     Milligram scale production of potent recombinant small interfering 
            RNAs in Escherichia coli.
  JOURNAL   Biotechnol Bioeng. 2018 Jun 5. doi: 10.1002/bit.26740.
  PUBMED    29873060
REFERENCE   2  (bases 1 to 7183)
  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"; journalName: "Biotechnol 
            Bioeng. 2018 Jun 5. doi: 10.1002/bit.26740."
COMMENT     Sequence Label: pET-pro-siRNA-V2-TP53
FEATURES             Location/Qualifiers
     source          1..7183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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)."
     RBS             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             73..90
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1293..1311
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2000..2018)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2025..2042
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2095..2113)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2109..2156
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2193..2648
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2280..2299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2490..2511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2740..3555)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2740..3555)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3463..3482
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3547..3566
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3677..4265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4166..4185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4419..4436
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4451..4590
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4516..4535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4676..4698
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4692..4883)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5692..6774)
                     /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     6736..6755
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6775..6852)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7058..7077
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7165..7183
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatatacata tgcatcatca ccaccatcac gaacgtgcaa tccagggcaa cgacgcccgt
      121 gaacaggcca acagcgaacg ctgggatggt ggcagcggcg gtacaaccag cccttttaaa
      181 ctgccggacg aaagcccgag ttggaccgaa tggcgcctgc ataacgatga gaccaacagc
      241 aaccaggaca acccgctggg ttttaaagaa agctggggct tcggcaaagt ggtgttcaag
      301 cgctatctgc gctacgatcg taccgaagcc agcctgcatc gcgttctggg tagctggacc
      361 ggtgacagcg tgaattatgc cgccagccgt tttttcggct tcgatcagat cggctgcacc
      421 tacagcattc gcttccgcgg tgtgagcatt accgttagcg gcggtagccg tacactgcag
      481 cacctgtgtg aaatggccat tcgcagtaag caagagctgc tgcagctggc cccgattgag
      541 gtggaaagta atgtgagccg tggctgtccg gagggcaccc aaaccttcga aaaagagagt
      601 gagtaatgat gaaccccatc gtaattaatc ggcttcaacg gaagctgggc tacactttta
      661 atcatcagga actgttgcag caggcattaa ctcatcgtag tgccagcagt aaacataacg
      721 agcgtttaga atttttaggc gactctattc tgagctacgt tatcgccaat gcgctttatc
      781 accgtttccc tcgtgtggat gaaggcgata tgagccggat gcgcgccacg ctggtccgtg
      841 gcaatacgct ggcggaactg gcgcgcgaat ttgagttagg cgagtgctta cgtttagggc
      901 caggtgaact taaaagcggt ggatttcgtc gtgagtcaat tctcgccgac accgtcgaag
      961 cattaattgg tggcgtattc ctcgacagtg atattcaaac cgtcgagaaa ttaatcctca
     1021 actggtatca aactcgtttg gacgaaatta gcccaggcga taaacaaaaa gatccgaaaa
     1081 cgcgcttgca agaatatttg cagggtcgcc atctgccgct gccgacttat ctggtagtcc
     1141 aggtacgtgg cgaagcgcac gatcaggaat ttactatcca ctgccaggtc agcggcctga
     1201 gtgaaccggt ggttggcaca ggttcaagcc gtcgtaaggc tgagcaggct gccgccgaac
     1261 aggcgttgaa aaaactggag ctggaatgaa tctaatacga ctcactatag gtgagctcta
     1321 ctcccctgcc ctcaacaaga tgttttgcca actggccaag acctgccctg tgcagctgtg
     1381 ggttgattcc acacccccgc ccggcacccg cgtccgcgcc atggccatct acaagcagtc
     1441 acagcacatg acggaggttg tgaggcgctg cccccaccat gagcgctgct cagatagcga
     1501 tggtctggcc cctcctcagc atcttatccg agtggaagga aatttgcgtg tggagtattt
     1561 ggatgacaga aacacttttc gacatagtgt ggtggtgccc tatgagccgc ctgaggttgc
     1621 tcgagttgac tattgcttaa cgatatacgt actcgtttaa atctctgatc atcgaatcta
     1681 aaagtcgacc aacctcaggc ggctcatagg gcaccaccac actatgtcga aaagtgtttc
     1741 tgtcatccaa atactccaca cgcaaatttc cttccactcg gataagatgc tgaggagggg
     1801 ccagaccatc gctatctgag cagcgctcat ggtgggggca gcgcctcaca acctccgtca
     1861 tgtgctgtga ctgcttgtag atggccatgg cgcggacgcg ggtgccgggc gggggtgtgg
     1921 aatcaaccca cagctgcaca gggcaggtct tggccagttg gcaaaacatc ttgttgaggg
     1981 caggggagta gcggccgccc ctatagtgag tcgtattagt cgagcaccac caccaccacc
     2041 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     2101 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     2161 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     2221 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2281 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2341 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2401 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2461 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2521 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2581 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     2641 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2701 taaatacatt caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat
     2761 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     2821 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     2881 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     2941 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     3001 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     3061 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     3121 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     3181 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     3241 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     3301 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     3361 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     3421 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     3481 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg
     3541 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     3601 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3661 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3721 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3781 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3841 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3901 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3961 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4021 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4081 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4141 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4201 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4261 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4321 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4381 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4441 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4501 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     4561 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     4621 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4681 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     4741 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     4801 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4861 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4921 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4981 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     5041 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     5101 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     5161 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     5221 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     5281 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     5341 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     5401 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     5461 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     5521 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     5581 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     5641 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     5701 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5761 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     5821 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     5881 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     5941 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     6001 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     6061 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     6121 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     6181 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     6241 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     6301 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     6361 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     6421 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     6481 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     6541 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     6601 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     6661 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     6721 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     6781 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     6841 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     6901 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     6961 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     7021 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     7081 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     7141 agatcgatct cgatcccgcg aaattaatac gactcactat agg
//
