LOCUS       sequence-8653-ex        8354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral Empty control vector, miRNA (CMV/TO), 3'LTR insertion, 
            Neo .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8354)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, 
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
  PUBMED    19657394
REFERENCE   2  (bases 1 to 8354)
  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: "PLoS One.";
            date: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT     Sequence Label: pQCXIN X2/pSM2(CMV/TO) empty (w334-1)
FEATURES             Location/Qualifiers
     source          1..8354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1604..1907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1908..2111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2108..2132
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    join(2290..2863,2864..2866,2867..2875)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2457..2474)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2684..2703
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2876..3670
                     /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="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2930..2949)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3540..3559
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    4153..4177
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     ncRNA           complement(4209..4321)
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           complement(4335..4428)
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     protein_bind    4466..4484
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4487..4505
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(4507..4710)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        4711..5014
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    complement(5037..5061)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5588..5610)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5746..5766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5751..6080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5931..6066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6102..6121
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6202..6219)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6373..6961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6453..6472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7132..7923,7924..7992))
                     /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     7755..7774
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7993..8097)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8165..8183
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8283..8354,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc tgggggatcg atcctctaga gtccgttaca taacttacgg
     1621 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     1681 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     1741 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     1801 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1861 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1921 ggcagtacat caatgggcgt gaatagcggt ttgactcacg gggatttcca agtctccacc
     1981 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2041 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2101 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     2161 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa
     2221 cgctgcagga attgatccgc ggccgcaccg gtaggcctcg tacgcttaat taacggatcc
     2281 ggaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata
     2341 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     2401 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     2461 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     2521 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     2581 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     2641 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     2701 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     2761 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaacgtct aggccccccg
     2821 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttgcca caaccatggc
     2881 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2941 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3001 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3061 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3121 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3181 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3241 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3301 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3361 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3421 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3481 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3541 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3601 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3661 gttcttctga gcgggatcgg ctagagtcga cccggacggg acgctcctgc gcctgataca
     3721 gaacgaattg cttgcaggca tctcatgagt gtgtcttccc gttttccgcc tgaggtcact
     3781 gcgtggatgg agcgctggcg cctgctgcgc gacggcgagc tgctcaccac ccactcgagg
     3841 gctgcagcgc tgcagaggcc gagtgcagaa ctgctccaaa gggacctcaa ggctttccga
     3901 gggacactag gctgactcca tcgagccagt gtagagataa gcttatcgat tagtccaatt
     3961 tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct
     4021 gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa
     4081 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gactcgagcg gccgccactg
     4141 tgctggatat caaccacttt gtacaagaaa gctgggtcta gataaacgca ttagtcttcc
     4201 aattgaaaaa agtgatttaa tttataccat tttaattcag ctttgtaaaa atgtatcaaa
     4261 gagatagcaa ggtattcagt tttagtaaac aagataattg ctcctaaagt agccccttga
     4321 attctggttg ctcgagcctt ctgttgggtt aacctgaaga agtaatccca gcaagtgttt
     4381 ccaagatgtg caggcaacga ttctgtaaag tactgaagcc tcattcaaac aattaccctg
     4441 ttatccctag tcgactagtc gacgatctct atcactgata gggagatctc tatcactgat
     4501 agggagagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     4561 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     4621 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     4681 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     4741 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     4801 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     4861 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     4921 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     4981 gccaggcggg ccatttaccg taagttatgt aacgcggaac tcccaggttc ctttaaagcc
     5041 tgcttttttg tacaaacttg ttgatatctg cagaattcca ccacactgga ctagcttaag
     5101 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa
     5161 ggtcaggaac agatggaaca gggtcgaccc tagagaacca tcagatgttt ccagggtgcc
     5221 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     5281 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     5341 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     5401 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     5461 tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca
     5521 gggaccaccg acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg
     5581 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     5641 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     5701 ccatgaccca gtcacgtagc gatagcggag tgtagatccg gctgtggaat gtgtgtcagt
     5761 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5821 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5881 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5941 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6001 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     6061 gaggcctagg cttttgcaaa aagcttactg gcttaactat gcggcatcag agcagattgt
     6121 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6181 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6781 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7021 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7081 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7141 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7201 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7261 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7321 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7381 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7441 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7501 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7561 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7621 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7681 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7741 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7801 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7861 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7921 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7981 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8041 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8101 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     8161 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttagcttggc cattgcatac
     8221 gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat taccgccatg
     8281 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     8341 cccatatatg gagt
//
