LOCUS       Exported                8623 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Retroviral Luciferase Expression vector, CMV/TO promoter, Blast .
ACCESSION   .
VERSION     .
KEYWORDS    pQCXIB CMV/TO LUC (w431-1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8623)
  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 8623)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8623
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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        1609..1912
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1913..2116
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    2118..2136
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2139..2157
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2198..2222
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2228..3880
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2294..2312)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    complement(3887..3911)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3964..4005
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4052..4624
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4219..4236)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4446..4465
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4683..5081
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5329..5754
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5857..5879)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6015..6035)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6020..6349
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6200..6335
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6262..6281
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6371..6390
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6471..6488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6642..7230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6722..6741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7401..8261)
                     /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     8024..8043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8262..8366)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8434..8452
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        8552..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 ataagcttgg gagttccgcg ttacataact
     1621 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     1681 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     1741 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     1801 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1861 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1921 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1981 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2041 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2101 ctatataagc agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg
     2161 tcgactagtc cagtgtggtg gaattctgca gatatcaaca agtttgtaca aaaaagcagg
     2221 ctccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga
     2281 agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg
     2341 aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga
     2401 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
     2461 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
     2521 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
     2581 gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa
     2641 cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta
     2701 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
     2761 atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc
     2821 ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag
     2881 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
     2941 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
     3001 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca
     3061 ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag
     3121 cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc
     3181 cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca
     3241 aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa
     3301 accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac
     3361 cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat
     3421 gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct
     3481 acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct
     3541 gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt
     3601 gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg
     3661 tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat
     3721 cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt
     3781 tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat
     3841 cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctagaccc agctttcttg
     3901 tacaaagtgg ttgatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc
     3961 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtaggcct
     4021 cgtacgctta attaacggat ccggaattcc gcccctctcc ctcccccccc cctaacgtta
     4081 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     4141 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     4201 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     4261 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     4321 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     4381 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     4441 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     4501 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     4561 taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     4621 ataagcttgc cacggggacg tggttttcct ttgaaaaaca cgatgataag cttgccacaa
     4681 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     4741 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     4801 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     4861 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     4921 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     4981 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     5041 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agcacttcgt ggccgaggag
     5101 caggactgac acgacccgga cgggacgctc ctgcgcctga tacagaacga attgcttgca
     5161 ggcatctcat gagtgtgtct tcccgttttc cgcctgaggt cactgcgtgg atggagcatc
     5221 agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga
     5281 gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     5341 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     5401 ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag ggtcgaccct
     5461 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     5521 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     5581 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     5641 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     5701 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
     5761 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
     5821 tggctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5881 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5941 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     6001 gtagatccgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     6061 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6181 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6241 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6301 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttactgg
     6361 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     6421 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     6481 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6541 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6601 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6661 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6721 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6781 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6841 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6901 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6961 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7021 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7081 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7141 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7201 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7261 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7321 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7381 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7441 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7501 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7561 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7621 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7681 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     7741 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7801 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7861 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7921 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7981 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     8041 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8101 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8161 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8221 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8281 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8341 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     8401 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     8461 cttcaagaat tagcttggcc attgcatacg ttgtatccat atcataatat gtacatttat
     8521 attggctcat gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag
     8581 taatcaatta cggggtcatt agttcatagc ccatatatgg agt
//
