LOCUS       sequence-314278-        7766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control lentiviral vector for mammalian gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7766)
  AUTHORS   Gstalder C, Liu D, Miao D, Lutterbach B, DeVine AL, Lin C, Shettigar
            M, Pancholi P, Buchbinder EI, Carter SL, Manos MP, Rojas-Rudilla V, 
            Brennick R, Gjini E, Chen PH, Lako A, Rodig S, Yoon CH, Freeman GJ, 
            Barbie DA, Hodi FS, Miles W, Van Allen EM, Haq R
  TITLE     Inactivation of Fbxw7 impairs dsRNA sensing and confers resistance 
            to PD-1 blockade.
  JOURNAL   Cancer Discov. 2020 May 5. pii: 2159-8290.CD-19-1416. doi: 
            10.1158/2159-8290.CD-19-1416.
  PUBMED    32371478
REFERENCE   2  (bases 1 to 7766)
  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: "Cancer 
            Discov. 2020 May 5. pii: 2159-8290.CD-19-1416. doi: 
            10.1158/2159-8290.CD-19-1416."
COMMENT     Sequence Label: pLX304_zeo_empty
FEATURES             Location/Qualifiers
     source          1..7766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..501
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     420..438
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             513..887
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_feature    947..1064
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1181..1484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1485..1688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1639..1659
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1711..1735
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1801..1825)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1827..1868
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1910..2498
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1963..1983)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2381..2392)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2570..2803
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2881..3002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2918..2937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2972..2991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3042..3177
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3197..3216)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3198..3216)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3226..3243)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3226..3242)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3235..3257)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3384..3839
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3471..3490)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3681..3702
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3865..3969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3970..4038,4039..4830)
                     /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(4188..4207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5001..5589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5490..5509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5743..5760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5877..5898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5913..5918,5919..5936,5937..5943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5951..5967
                     /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     5956..5978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5975..5991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5975..5991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6010..6030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6012..6030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6058..6284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6285..6465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6512..6637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7130..7363
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7548..7592
                     /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"
ORIGIN
        1 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
       61 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
      121 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
      181 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
      241 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
      301 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
      361 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
      421 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
      481 atcaccgacc tctctcccca gggggtacca ccatggccaa gttgaccagt gccgttccgg
      541 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
      601 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
      661 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
      721 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
      781 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
      841 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgactg cagctgcagt
      901 agtaagaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
      961 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     1021 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     1081 acagcagaga tccactttgg cgccggctcg agggggcccg gggttattaa tagtaatcaa
     1141 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     1201 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     1261 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     1321 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1381 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     1441 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     1501 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     1561 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     1621 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1681 gcagagctct ctggctaact gtcgggatca acaagtttgt acaaaaaagc aggctttaaa
     1741 ggaaccaatt cagtcgactg gatccggtac cgaattcgcg gccgcactcg agatatctag
     1801 acccagcttt cttgtacaaa gtggttggta agcctatccc taaccctctc ctcggtctcg
     1861 attctacgta gtaatgagct agcgctaacc ggtggcgcgt taagtcgaca atcaacctct
     1921 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     1981 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2041 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2101 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2161 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2221 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2281 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     2341 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2401 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2461 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
     2521 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     2581 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     2641 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     2701 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     2761 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca
     2821 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     2881 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2941 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3001 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     3061 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3121 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3181 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     3241 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3301 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     3361 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3421 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3481 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3541 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3601 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3661 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3721 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3781 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3841 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3901 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3961 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4021 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4081 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4141 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4201 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4261 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4321 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4381 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4441 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4501 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4561 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4621 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4681 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4741 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4801 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4861 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4921 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4981 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcataatctg ctgcttgcaa
     5041 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5101 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5161 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5221 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5281 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5341 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5401 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5461 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5521 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5581 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5641 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5701 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5761 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5821 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5881 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     5941 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6001 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
     6061 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
     6121 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
     6181 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
     6241 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
     6301 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6361 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6421 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     6481 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     6541 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     6601 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     6661 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     6721 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     6781 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     6841 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     6901 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     6961 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     7021 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     7081 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     7141 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     7201 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     7261 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     7321 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     7381 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     7441 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     7501 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     7561 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     7621 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     7681 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     7741 cccacctccc aaccccgagg ggaccc
//
