LOCUS       sequence-291325-        6944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsZNF598-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6944)
  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"
COMMENT     Sequence Label: pT7-V5-SBP-C1-HsZNF598-siRNAres_AH
FEATURES             Location/Qualifiers
     source          1..6944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    3732..3853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3769..3788)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3823..3842
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3860..4315)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3997..4018)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4209..4228
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4342..4446
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4448..4805
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4448..4639
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4471..4491)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4656..4791
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4840..5634
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4840..5634
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4894..4913)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5504..5523
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5822..5841)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5866..5913
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6242..6830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6731..6750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagctatgg cggcggcggg gggcgccgag gggcggcgcg cggccctgga
      901 ggcggcggcg gcggcagctc ctgagcgggg aggcgggagc tgcgtgctgt gctgcggaga
      961 cctggaggcc acggcgctgg gccgctgcga ccacccggtg tgctaccgct gctctaccaa
     1021 gatgcgggtg ctctgcgagc agcgctactg cgccgtgtgc cgcgaggagc tgcgccaggt
     1081 ggtctttggg aagaagcttc ctgcctttgc caccatcccc atccaccagc tgcagcatga
     1141 gaagaaatat gatatctact ttgcagatgg aaaggtgtac gcattgtaca ggcagctgct
     1201 gcagcacgag tgcccgcggt gccccgagct gccacctttc agcctcttcg gggacctgga
     1261 gcagcacatg cggaggcagc atgagctctt ctgctgccgt ctgtgcctcc agcacctcca
     1321 gatcttcaca tatgagcgca agtggtactc gcgcaaggac ctggcccggc atcgcatgca
     1381 gggtgacccc gatgacacgt cgcaccgtgg gcacccgctc tgcaagttct gtgacgagcg
     1441 ctacctggac aatgatgagc tgcttaagca cctgcgccgc gaccactact tctgccactt
     1501 ctgcgactcg gacggggccc aggactacta cagcgactat gcctacctgc gtgagcactt
     1561 ccgggagaag cactttctgt gtgaggaagg ccgctgcagc acggagcagt tcacccacgc
     1621 cttccgcacc gagatcgacc tcaaggccca caggacggcc tgccacagtc gcagccgcgc
     1681 cgaggcacgc cagaaccgcc acatcgacct gcagttcagc tacgcgccac ggcactcgcg
     1741 ccggaacgag ggggtcgttg gtggcgaaga ctacgaggag gtggacaggt acagccgcca
     1801 gggccgagtg gcccgggctg gcactcgcgg agcccagcag agccgccgag gaagctggag
     1861 gtacaaaagg gaagaagagg accgagaagt agcagctgct gtccgggcct ccgtggccgc
     1921 acagcagcag gaggaggccc gaagaagcga agaccaggag gaaggcggta ggccaaaaaa
     1981 ggaagaagcg gcggcgcggg gacctgagga tccccgtggc ccccggcgct caccccggac
     2041 tcagggcgaa ggcccaggcc ccaaggaaac ctcgacaaat ggtcctgtaa gccaagaagc
     2101 cttctcggtg acaggcccag ccgccccagg gtgtgtgggg gtgccaggcg ccctcccacc
     2161 acccagcccg aagctcaagg acgaagactt ccccagcctc tctgcctcca cttcctcctc
     2221 ctgctccact gcagcaaccc cgggccctgt ggggttggcg ctgccgtacg ccatccctgc
     2281 cagaggcagg agtgccttcc aggaggagga cttccccgcc ctggtgtcct cggtgcccaa
     2341 gcctggcacc gcccccacca gccttgtctc tgcctggaac agcagcagta gcagcaagaa
     2401 ggtagcacag cccccactct cggcgcaggc taccggcagc ggccagccca ccaggaaggc
     2461 tgggaagggg agcaggggcg gcaggaaggg cggcccgccc ttcacacagg aggaggagga
     2521 ggacggcggc ccggccctgc aggagcttct gagcacacgc cccacgggct ccgtctcctc
     2581 cacactgggg ctggcctcca tccaaccctc taaagttggg aagaagaaga aagtgggctc
     2641 ggagaagcca ggcaccacat tgccacagcc cccgcccgct acctgtcccc caggggcttt
     2701 gcaggccccg gaagctcctg ccagcagagc cgaggggcca gttgccgtcg tcgttaatgg
     2761 acacacggag ggcccggccc ctgctcggag tgcccccaag gaaccccctg ggctcccaag
     2821 gcccctgggg tccttcccct gccccacgcc acaggaggac ttcccagcgc tcggcggccc
     2881 ctgcccaccc cggatgccgc cgcccccagg cttcagcgct gtggtgctcc tgaagggcac
     2941 gcctccccca cccccgccgg gcctggtgcc cccaatcagc aagccgcccc ctggcttctc
     3001 tggccttctg cctagccccc acccggcctg tgtccccagc cccgccacca ccaccaccac
     3061 aaaagcaccc aggctgctgc ctgccccacg ggcgtaccta gtccccgaga acttccggga
     3121 gaggaacctt cagctcatcc agtccatcag ggacttcctg cagagcgacg aggcccgctt
     3181 cagcgagttc aagagccact caggggagtt cagacagggc ctgatctccg cagcccagta
     3241 ttacaagagt tgccgggacc tgctggggga gaatttccag aaggtcttta atgagctgct
     3301 ggtcctgctg cccgacacgg ccaagcagca ggagctcctg tctgcacaca cggacttctg
     3361 caaccgcgag aagcctctga gcaccaagtc caagaagaac aagaagagcg cgtggcaggc
     3421 caccacccag caggcgggcc tggactgccg tgtgtgcccc acctgccagc aggtgctcgc
     3481 gcatggcgat gccagcagcc accaggcgct gcatgctgcc cgggacgacg acttcccctc
     3541 cctgcaagcc atcgccagga tcatcacgta ggaattctgc agtcgacggt accgcgggcc
     3601 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     3661 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3721 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3781 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3841 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3901 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3961 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4021 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4081 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4141 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4201 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4261 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4321 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4381 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4441 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4501 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4561 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4621 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4681 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4741 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4801 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4861 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4921 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4981 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     5041 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5101 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5161 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5221 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5281 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5341 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5401 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5461 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5521 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5581 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5641 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5701 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5761 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5821 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5881 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5941 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     6001 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     6061 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     6121 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6181 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6241 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6301 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6361 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6421 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6481 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6541 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6601 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6661 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6721 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6781 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6841 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6901 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
