LOCUS       sequence-239544-        7754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd gen lentiviral empty control vector, CMV promoter, Blast .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7754)
  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 7754)
  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: pLenti CMV Blast empty (w263-1)
FEATURES             Location/Qualifiers
     source          1..7754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..22
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    139..160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(175..180,181..198,199..205)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    213..229
                     /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)."
     primer_bind     218..240
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     237..253
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     237..253
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     272..292
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        274..292
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        320..546
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             547..727
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    774..899
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1392..1625
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1810..1854
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2121..2237
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2260..2563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2564..2767
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2718..2738
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2764..2788
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2872..2896
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2965..2989)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3020..3608
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3073..3093)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3491..3502)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3659..3679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3664..3993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3844..3979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4041..4088
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4107..4505
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4592..4825
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4897..5018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4934..4953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4988..5007
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5058..5193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5213..5232)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5214..5232)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5242..5259)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5242..5258)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5251..5273)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5400..5855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5487..5506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5697..5718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5881..5985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5986..6054,6055..6846)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6204..6223)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7017..7605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7506..7525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
       61 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      121 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      181 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      241 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
      301 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
      361 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
      421 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
      481 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
      541 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      601 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
      661 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
      721 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
      781 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
      841 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
      901 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
      961 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1021 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1081 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1141 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1201 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     1261 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     1321 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     1381 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     1441 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     1501 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     1561 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     1621 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     1681 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     1741 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     1801 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     1861 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     1921 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     1981 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     2041 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     2101 tctcgacggt atcggttaac ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     2161 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     2221 acaaaattca aaattttatc gataagcttg ggagttccgc gttacataac ttacggtaaa
     2281 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     2341 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     2401 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     2461 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     2521 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     2581 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     2641 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     2701 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     2761 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     2821 ccatagaaga caccgactct agtccagtgt ggtggaattc tgcagatatc aacaagtttg
     2881 tacaaaaaag caggctccac catgggaacc aattcagtcg actggatccg gtaccgaatt
     2941 cgcggccgca ctcgagatat ctagacccag ctttcttgta caaagtggtt gatatccagc
     3001 acagtggcgg ccgctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3061 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3121 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3181 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3241 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3301 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3361 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3421 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3481 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3541 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3601 ctccccgcct ggaattctgc agatatccgg ttagtaatga gtttggaatt aattctgtgg
     3661 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3721 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3781 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3841 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3901 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3961 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4021 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     4081 aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa gaatccaccc
     4141 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     4201 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     4261 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     4321 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     4381 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     4441 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     4501 gctaagcaca attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat
     4561 cttagccact ttttaaaaga aaagggggga ctggaagggc tagctcactc ccaacgaaga
     4621 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     4681 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     4741 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     4801 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     4861 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     4921 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4981 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     5041 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5101 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     5161 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     5221 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     5281 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     5341 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     5401 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5461 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5521 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5581 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5641 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5701 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5761 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5821 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     5881 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5941 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6001 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6061 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6121 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6181 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6241 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6301 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6361 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6421 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6481 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6541 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6601 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6661 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6721 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6781 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6841 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6901 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6961 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7141 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7321 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7381 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7441 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7501 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7561 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7621 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7681 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7741 agccgaacga ccga
//
