LOCUS       sequence-237311-        7607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET-pro-siRNA-V2 based plasmid for production of recombinant siRNAs 
            (pro-siRNAs) against human Lamin A/C gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7607)
  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 7607)
  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-LMNA
FEATURES             Location/Qualifiers
     source          1..7607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             34..51
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1254..1272
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2385..2403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2410..2427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2480..2498)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2494..2541
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2578..3033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2665..2684)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2875..2896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3125..3940)
                     /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(3125..3940)
                     /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     3848..3867
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3932..3951
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4062..4650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4551..4570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4804..4821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4836..4975
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4901..4920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5061..5083
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5077..5268)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6077..7159)
                     /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     7121..7140
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7160..7237)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7443..7462
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7550..7568
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7569..7593
                     /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)."
ORIGIN
        1 tttgtttaac tttaagaagg agatatacat atgcatcatc accaccatca cgaacgtgca
       61 atccagggca acgacgcccg tgaacaggcc aacagcgaac gctgggatgg tggcagcggc
      121 ggtacaacca gcccttttaa actgccggac gaaagcccga gttggaccga atggcgcctg
      181 cataacgatg agaccaacag caaccaggac aacccgctgg gttttaaaga aagctggggc
      241 ttcggcaaag tggtgttcaa gcgctatctg cgctacgatc gtaccgaagc cagcctgcat
      301 cgcgttctgg gtagctggac cggtgacagc gtgaattatg ccgccagccg ttttttcggc
      361 ttcgatcaga tcggctgcac ctacagcatt cgcttccgcg gtgtgagcat taccgttagc
      421 ggcggtagcc gtacactgca gcacctgtgt gaaatggcca ttcgcagtaa gcaagagctg
      481 ctgcagctgg ccccgattga ggtggaaagt aatgtgagcc gtggctgtcc ggagggcacc
      541 caaaccttcg aaaaagagag tgagtaatga tgaaccccat cgtaattaat cggcttcaac
      601 ggaagctggg ctacactttt aatcatcagg aactgttgca gcaggcatta actcatcgta
      661 gtgccagcag taaacataac gagcgtttag aatttttagg cgactctatt ctgagctacg
      721 ttatcgccaa tgcgctttat caccgtttcc ctcgtgtgga tgaaggcgat atgagccgga
      781 tgcgcgccac gctggtccgt ggcaatacgc tggcggaact ggcgcgcgaa tttgagttag
      841 gcgagtgctt acgtttaggg ccaggtgaac ttaaaagcgg tggatttcgt cgtgagtcaa
      901 ttctcgccga caccgtcgaa gcattaattg gtggcgtatt cctcgacagt gatattcaaa
      961 ccgtcgagaa attaatcctc aactggtatc aaactcgttt ggacgaaatt agcccaggcg
     1021 ataaacaaaa agatccgaaa acgcgcttgc aagaatattt gcagggtcgc catctgccgc
     1081 tgccgactta tctggtagtc caggtacgtg gcgaagcgca cgatcaggaa tttactatcc
     1141 actgccaggt cagcggcctg agtgaaccgg tggttggcac aggttcaagc cgtcgtaagg
     1201 ctgagcaggc tgccgccgaa caggcgttga aaaaactgga gctggaatga atctaatacg
     1261 actcactata ggtgagctcc aagaccaaga cccttgactc agtagccaag gagcgcgccc
     1321 gcctgcagct ggagctgagc aaagtgcgtg aggagtttaa ggagctgaaa gcgcgcaata
     1381 ccaagaagga gggtgacctg atagctgctc aggctcggct gaaggacctg gaggctctgc
     1441 tgaactccaa ggaggccgca ctgagcactg ctctcagtga gaagcgcacg ctggagggcg
     1501 agctgcatga tctgcggggc caggtggcca agcttgaggc agccctaggt gaggccaaga
     1561 agcaacttca ggatgagatg ctgcggcggg tggatgctga gaacaggctg cagaccatga
     1621 aggaggaact ggacttccag aagaacatct acagtgagga gctgcgtgag accaagcgcc
     1681 gtcatgagac ccgactggtg gagattgaca atgggaagca gcgtgagttt gagagccggc
     1741 tggcggatgc gctgcaggaa ctgcgggccc agcatgagga ccaggtggag cagtataaga
     1801 aggagctgct cgagtctaga gggcccgttt aaacccgctg atcagcgtcg accagctcct
     1861 tcttatactg ctccacctgg tcctcatgct gggcccgcag ttcctgcagc gcatccgcca
     1921 gccggctctc aaactcacgc tgcttcccat tgtcaatctc caccagtcgg gtctcatgac
     1981 ggcgcttggt ctcacgcagc tcctcactgt agatgttctt ctggaagtcc agttcctcct
     2041 tcatggtctg cagcctgttc tcagcatcca cccgccgcag catctcatcc tgaagttgct
     2101 tcttggcctc acctagggct gcctcaagct tggccacctg gccccgcaga tcatgcagct
     2161 cgccctccag cgtgcgcttc tcactgagag cagtgctcag tgcggcctcc ttggagttca
     2221 gcagagcctc caggtccttc agccgagcct gagcagctat caggtcaccc tccttcttgg
     2281 tattgcgcgc tttcagctcc ttaaactcct cacgcacttt gctcagctcc agctgcaggc
     2341 gggcgcgctc cttggctact gagtcaaggg tcttggcggc cgcccctata gtgagtcgta
     2401 ttagtcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     2461 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     2521 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     2581 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2641 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2701 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2761 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2821 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2881 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2941 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3001 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     3061 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     3121 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3181 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3241 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     3301 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3361 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     3421 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     3481 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     3541 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     3601 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     3661 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     3721 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3781 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     3841 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     3901 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     3961 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     4021 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4081 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4141 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4201 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4261 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4321 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4381 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4441 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4501 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4561 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4621 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4681 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4741 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4801 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4861 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     4921 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     4981 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5041 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5101 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     5161 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     5221 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     5281 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     5341 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     5401 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     5461 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     5521 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     5581 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     5641 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     5701 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     5761 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     5821 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     5881 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     5941 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     6001 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     6061 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6121 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     6181 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     6241 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     6301 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     6361 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     6421 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     6481 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     6541 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     6601 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     6661 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     6721 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     6781 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     6841 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     6901 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     6961 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     7021 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     7081 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     7141 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     7201 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     7261 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     7321 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     7381 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     7441 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     7501 acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt aatacgactc
     7561 actatagggg aattgtgagc ggataacaat tcccctctag aaataat
//
