LOCUS       sequence-212775-        9231 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid for Env from the Q769.d22 HIV-1 
            isolate; QES mutant with enhanced binding to antibody PG16.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9231)
  AUTHORS   Heredia JD, Park J, Choi H, Gill KS, Procko E
  TITLE     Conformational engineering of HIV-1 Env based on mutational 
            tolerance in the CD4 and PG16 bound states.
  JOURNAL   J Virol. 2019 Mar 20. pii: JVI.00219-19. doi: 10.1128/JVI.00219-19.
  PUBMED    30894475
REFERENCE   2  (bases 1 to 9231)
  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: "J Virol. 
            2019 Mar 20. pii: JVI.00219-19. doi: 10.1128/JVI.00219-19."
COMMENT     Sequence Label: pCEP4-Q769gp160-QES.i03.V255M
FEATURES             Location/Qualifiers
     source          1..9231
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        86..231
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             274..1311
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     primer_bind     complement(1309..1328)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1353..1401
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        2055..2434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2435..2638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2589..2609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2635..2659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     RBS             4323..4331
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5277..5411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5301..5320)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5355..5374
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5554..5576)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      6136..7925
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
ORIGIN
        1 gccctgcttc atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat
       61 atatttgcat gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc
      121 gaattcgaac acgcagatgc agtcggggcg gcgcggtccg aggtccactt cgcatattaa
      181 ggtgacgcgt gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa
      241 cagcgtgccg cagatcccgg ggggcaatga gatatgaaaa agcctgaact caccgcgacg
      301 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
      361 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
      421 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
      481 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
      541 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
      601 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
      661 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
      721 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
      781 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
      841 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
      901 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
      961 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     1021 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     1081 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     1141 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     1201 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     1261 aaccgacgcc ccagcactcg tccggatcgg gagatggggg aggctaactg aaacacggaa
     1321 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     1381 cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     1441 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     1501 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     1561 ccatagccac tggccccgtg ggttagggac ggggtccccc atggggaatg gtttatggtt
     1621 cgtgggggtt attattttgg gcgttgcgtg gggtcaggtc cacgactgga ctgagcagac
     1681 agacccatgg tttttggatg gcctgggcat ggaccgcatg tactggcgcg acacgaacac
     1741 cgggcgtctg tggctgccaa acacccccga cccccaaaaa ccaccgcgcg gatttctggc
     1801 gtgccaagct agtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac
     1861 gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata cattgaatca
     1921 atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg
     1981 gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat
     2041 atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     2101 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2161 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2221 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2281 cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg
     2341 taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca
     2401 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa
     2461 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2521 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc
     2581 cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     2641 tttagtgaac cgtcagatct ctagaagctg ggtaccagct gctagcgcca ccatgcccat
     2701 ggggtctttg caacccctcg ctactcttta tttgctgggt atgttggtag cgagcgttct
     2761 ggcggcgggc aatctctggg taaccgtgta ctacggcgtg ccagtgtggc gggatgctga
     2821 aacaaccctc ttttgtgcaa gtgatgcaaa agcgtatgac cgagaagcac acaatgtttg
     2881 ggctacacat gcgtgcgttc cgacagatcc atccccccaa gaagtaccat tggggaatgt
     2941 aaccgaggaa ttcaacatgt ggaagaataa catggttgaa cagatgcata cagacatcat
     3001 ctctttgtgg gaccagtcac tgcagccgtg tgtgaaactt acgcctcttt gtgtcacgct
     3061 gaattgttcc aatagtaata acattccaag tgtcagcaac attaccgatg atatgaaaga
     3121 agagattaag aactgcagct ttaacatgac aaccgaattg aaggataaaa agcagaacgt
     3181 ttactctttg ttctatcgac tggacgttgt acccctcgaa acaaaaacca accagaacag
     3241 tagtcactca aggtatcggc tcattaactg caacacgtct gagatcacgc aggcttgccc
     3301 aaaagtttct ttcgagccga tccctattca ctattgtgcg cctgcaggct atgctatcct
     3361 gaaatgtaac gacaagggct ttaacggcac tggactttgt aaaaacgtct caacagtaca
     3421 atgcacgcat gggattaaac cggtcatgag tacccagctg ctccttaatg gaagccttgc
     3481 cgagggtaag gtaatggtac gatccgaaaa tattacaaat aatgcgaaaa acattataat
     3541 acagtttaac aactctgtgc aaattaattg cacgcgccca ggtaataaca cgcgaaaaag
     3601 tatacatctc gggcctggga aagtttttta cgcgacggac ataatagggg atataaggaa
     3661 agcgcactgt aatgttaatc gccaacaatg gaataagacc cttcaggatg tggcaacgca
     3721 actcaggacc cacttcagaa accgtacgat catatttaat aatagtctcg gtggggattt
     3781 ggaaataact acgcatagtt ttaattgtag gggagagttt ttctattgta acacttccgg
     3841 gctctttaat gggatatgga acggaacgca agaacctaat cggaccgagt ctaacgatac
     3901 aattacgctt caatgtcgga taaagcagat catcaacatg tggcagagag taggccaggc
     3961 aatttacgca ccccctatcc agggggagat cagatgcgaa agtaacataa ccggacttat
     4021 cctgacgagg gatgggggga tcatcaacag tacagaggaa acgtttagac ctgggggcgg
     4081 ggacatgaga gataattgga ggtccgagct ctataaatac aaagttgtaa agatagaacc
     4141 gcttggagtg gctccaacta aggccaagag acgggtcgtg gagagagaaa aacgggcggt
     4201 aggatttggc gcgttctttc tcggtttcct tggcgcagcc ggatcaacga tgggggccgc
     4261 ctccatcacc ctgacggttc aggcgcgcca gcttttgagt gggattgtgc agcagcagaa
     4321 taacctcctt agggccatag aggctcagca gcacctgctg aagcttaccg tttggggcat
     4381 taagcaactg caggcgcggg tactcgcggt tgaaagatac cttaaggatc agcaactgct
     4441 gggaatgtgg ggatgtagtg gaaagttcat ctgcacaacc acggtcccat ggaattcctc
     4501 atggtcaaat aaatcacagt cagaaatatg ggataatatg acgtggatgc agtgggacaa
     4561 ggagattaac aattacaccc agattatcta cgatcttatt gaggagtcac aaaggcaaca
     4621 agagaaaaac gaacaggact tgctggcgct cgacaaatgg gctaatttgt ggaactggtt
     4681 cgacatcagt aactggctgt ggtacataaa gattttcata atgatagttg gcgggctcat
     4741 agggcttcgg attgcgttcg cagtgctttc tgtcatcaat agggtcaggc aaggttattc
     4801 cccgctctcc tttcaaaccc ataccccaaa ccctcgggat ttggataggc caggccggat
     4861 agaggaggaa ggaggtgagc aggatcggga ccgaagcata agactggtct ctgggtttct
     4921 tgctttggca tgggacgacc tgaggtctct gtgtttgttc agttaccacc gcctgcgaga
     4981 tttcatcttg gtggctgcta gaacggtcga acttcttgga catatttcac ttaaaggctt
     5041 gagacgaggc tgggaaggat tgaaatacct tgggaacctt ctgtcatact ggggacggga
     5101 actgaagata agtgccataa acttgttgga cacaatcgcg attgttgtgg cggaatggac
     5161 cgatcggata attgagattg gccagagact gtgtcgcgct atcattaata tacctcggag
     5221 aatccgacag gggttcgaac gggctttgtt gtagtaactc gaggccggca aggccggatc
     5281 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     5341 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     5401 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     5461 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatccgg
     5521 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5581 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5641 gggtgttggc gggtgtcggg gcgcagccat gaggtcgact ctagaggatc gatccccgcc
     5701 ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc agtgcatgta
     5761 atcccttcag ttggttggta caacttgcca actgggccct gttccacatg tgacacgggg
     5821 ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa tggatgtgca
     5881 catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg tggactgcaa
     5941 cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg ctgggggaca
     6001 tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa tcagaggggc
     6061 ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt ttataaggcc
     6121 cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat actatccaga
     6181 ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc gaattagggt
     6241 tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc acggttttat
     6301 ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag ggcagtggct
     6361 gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt cattctcctt
     6421 cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga
     6481 aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc agtggtggca
     6541 ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata ctagtgtagg
     6601 aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac aagccgtaaa
     6661 gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc ctagtgcaat
     6721 atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg aaccatgttg
     6781 ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg gactccactg
     6841 gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc ccataaacaa
     6901 agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca
     6961 cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg attgtaaccc
     7021 cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc ccggggaata
     7081 cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc tggaggacaa
     7141 attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat attcacgagg
     7201 tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc aaatatctgg
     7261 atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     7321 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     7381 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     7441 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat agagattagg
     7501 gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat atctggatag
     7561 catatgctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     7621 cataggctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     7681 tatatgctat cctaatttat atctgggtag cataggctat cctaatctat atctgggtag
     7741 catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag
     7801 catatgctat cctcatgcat atacagtcag catatgatac ccagtagtag agtgggagtg
     7861 ctatcctttg catatgccgc cacctcccaa gggggcgtga attttcgctg cttgtccttt
     7921 tcctgctggt tgctcccatt cttaggtgaa tttaaggagg ccaggctaaa gccgtcgcat
     7981 gtctgattgc tcaccaggta aatgtcgcta atgttttcca acgcgagaag gtgttgagcg
     8041 cggagctgag tgacgtgaca acatgggtat gcccaattgc cccatgttgg gaggacgaaa
     8101 atggtgacaa gacagatggc cagaaataca ccaacagcac gcatgatgtc tactggggat
     8161 ttattcttta gtgcggggga atacacggct tttaatacga ttgagggcgt ctcctaacaa
     8221 gttacatcac tcctgccctt cctcaccctc atctccatca cctccttcat ctccgtcatc
     8281 tccgtcatca ccctccgcgg cagccccttc caccataggt ggaaaccagg gaggcaaatc
     8341 tactccatcg tcaaagctgc acacagtcac cctgatattg caggtaggag cgggctttgt
     8401 cataacaagg tccttaatcg catccttcaa aacctcagca aatatatgag tttgtaaaaa
     8461 gaccatgaaa taacagacaa tggactccct tagcgggcca ggttgtgggc cgggtccagg
     8521 ggccattcca aaggggagac gactcaatgg tgtaagacga cattgtggaa tagcaagggc
     8581 agttcctcgc cttaggttgt aaagggaggt cttactacct ccatatacga acacaccggc
     8641 gacccaagtt ccttcgtcgg tagtcctttc tacgtgactc ctagccagga gagctcttaa
     8701 accttctgca atgttctcaa atttcgggtt ggaacctcct tgaccacgat gctttccaaa
     8761 ccaccctcct tttttgcgcc tgcctccatc accctgaccc cggggtccag tgcttgggcc
     8821 ttctcctggg tcatctgcgg ggccctgctc tatcgctccc gggggcacgt caggctcacc
     8881 atctgggcca ccttcttggt ggtattcaaa ataatcggct tcccctacag ggtggaaaaa
     8941 tggccttcta cctggagggg gcctgcgcgg tggagacccg gatgatgatg actgactact
     9001 gggactcctg ggcctctttt ctccacgtcc acgacctctc cccctggctc tttcacgact
     9061 tccccccctg gctctttcac gtcctctacc ccggcggcct ccactacctc ctcgaccccg
     9121 gcctccacta cctcctcgac cccggcctcc actgcctcct cgaccccggc ctccacctcc
     9181 tgctcctgcc cctcctgctc ctgcccctcc tcctgctcct gcccctcctg c
//
