LOCUS       sequence-212706-        8845 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid for extracellular gp140 from the BaL 
            HIV-1 strain fused to a transmembrane tether.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8845)
  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 8845)
  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-BaLgp140-TM
FEATURES             Location/Qualifiers
     source          1..8845
                     /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"
     CDS             4634..4678
                     /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"
     CDS             4718..4735
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4878..5012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4902..4921)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4956..4975
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5155..5177)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      5737..7526
                     /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 ggggagcttg caaccgctcg ccactctcta tctcctcggc atgttggtag catctgtgct
     2761 ggctgaggaa aagctctggg tgactgtgta ttacggggtc ccagtttgga aggaggctac
     2821 taccacgctc ttctgcgcct ccgaccggaa ggcatacgac accgaggtgc ataatgtgtg
     2881 ggccacacac gcctgtgtgc ctaccgaccc caacccacag gaagtggaac tgaagaacgt
     2941 gactgagaat ttcaatatgt ggaaaaataa tatggtggag cagatgcacg aggacattat
     3001 tagcctctgg gatcagtccc ttaaaccctg cgtgaaactg acaccgctgt gcgtaacgct
     3061 gaattgtact gacctccgga atgccacgaa cggaaacgac acaaatacca cgtcaagctc
     3121 ccgagggatg gtgggagggg gagaaatgaa gaactgctca ttcaatataa cgacgaatat
     3181 ccgagggaag gttcagaaag agtatgccct gttttacaaa ctggacatcg ctcctatcga
     3241 caacaacagc aataacaggt accgcctgat tagctgcaac acttctgtga ttacccaggc
     3301 ctgccccaag gtgagcttcg agcctatccc aatccactac tgcgccccag caggctttgc
     3361 catcctgaaa tgcaaggaca agaagttcaa cggcaaaggc ccatgtacaa atgtcagtac
     3421 agtacagtgc acacacggta tccgcccagt tgtctctaca caactgctgc tgaatggatc
     3481 cttggcagag gaagaggtgg tgatcagatc tgctaacttc gccgataatg ctaaagtcat
     3541 cattgttcag ctgaacgagt ctgtcgaaat caattgtact cgcccaaaca acaacacgcg
     3601 caagagcatt catatcggac cgggcagggc cttttacaca acaggggaga ttattggtga
     3661 tattagacaa gcccactgca acctcagtcg agcaaagtgg aatgatactc tcaacaaaat
     3721 tgttatcaaa ttgcgggaac agtttgggaa caagacaatc gtatttaagc actcatccgg
     3781 aggcgatcca gaaatcgtga cacactcttt taactgcggt ggagaatttt tctactgcaa
     3841 ctccacccag ctgttcaaca gtacgtggaa cgtgacagag gagtctaaca acaccgtgga
     3901 gaataatacg atcacccttc catgtcggat taagcagatc atcaatatgt ggcaggaggt
     3961 gggaagagca atgtacgcgc cgcccattag gggccagatc agatgcagtt caaatatcac
     4021 aggcctgctc ctgacacgcg acggaggccc agaggacaac aagacagagg tgtttaggcc
     4081 cggcggcggc gacatgcgag ataattggag gtctgaattg tacaagtaca aggtggtgaa
     4141 aatcgaaccc ctgggagtgg cgcctactaa ggcaaagcgc cgcgttgtgc agagagagaa
     4201 gagggcagtg ggaattgggg cggtttttct tgggttcctg ggggctgcag gtagcaccat
     4261 gggagcggcc gctatgacac tgacagtgca agccagactc ctgctgtctg gtattgtcca
     4321 acagcaaaac aatttgctcc gcgcaataga agcccagcag cacctgctcc aactgactgt
     4381 gtggggcatt aaacaattgc aggctcgcgt gctggcagtc gaacggtacc tgagggatca
     4441 acagctcctg ggcatctggg gctgctccgg gaaactcatc tgtacgacag cggtcccttg
     4501 gaatgccagc tggtccaaca aaagcctcaa caagatttgg gacaatatga cgtggattga
     4561 gtgggacagg gaaattaaca attatacctc aatcatatat tctctgattg aagagtccca
     4621 gaatcagcaa gagaagaatg agcaggaatt gttggaactt gataaatggg caagcctgtg
     4681 gaactggttt gacataacca aatccggagg cagcggacat catcaccatc accacggctc
     4741 tggagccgga agtgctatcc ccatcatggg tatcgttgct ggcctggttg tccttgcagc
     4801 tgtagtcact ggagctgcgg tcgctgctgt gctgtggaga aagaagagct cagattgact
     4861 cgaggccggc aaggccggat ccagacatga taagatacat tgatgagttt ggacaaacca
     4921 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4981 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     5041 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     5101 gtatggctga ttatgatccg gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     5161 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5221 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgaggtcgac
     5281 tctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc tctgcccctt
     5341 cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc aactgggccc
     5401 tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac tgtagttgac
     5461 atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc tggagcagac
     5521 tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg gctgaagctc
     5581 ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac gggaggctac
     5641 accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca agagggcatt
     5701 agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat gcttcccggg
     5761 tagtagtata tactatccag actaacccta attcaatagc atatgttacc caacgggaag
     5821 catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata tctcccaccc
     5881 catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta gtgaaccatt
     5941 ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc
     6001 gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt gtgaacagta
     6061 aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg
     6121 acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac aaaccccttg
     6181 ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat agaaatccat
     6241 ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat ggctatgggc
     6301 aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc aggcagggac
     6361 caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga gagtggacgc
     6421 cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac ggggctccac
     6481 gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat tgtggagtgg
     6541 gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat aagggtgtaa
     6601 taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc cacaaaacca
     6661 ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat aggggcgcga
     6721 ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag cacagggttg
     6781 ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc catgggtagc
     6841 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatag
     6901 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     6961 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     7021 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     7081 gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg ggtagcatat
     7141 actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta gcatatgcta
     7201 tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta
     7261 tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta
     7321 tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta
     7381 tcctaatctg tatccgggta gcatatgcta tcctcatgca tatacagtca gcatatgata
     7441 cccagtagta gagtgggagt gctatccttt gcatatgccg ccacctccca agggggcgtg
     7501 aattttcgct gcttgtcctt ttcctgctgg ttgctcccat tcttaggtga atttaaggag
     7561 gccaggctaa agccgtcgca tgtctgattg ctcaccaggt aaatgtcgct aatgttttcc
     7621 aacgcgagaa ggtgttgagc gcggagctga gtgacgtgac aacatgggta tgcccaattg
     7681 ccccatgttg ggaggacgaa aatggtgaca agacagatgg ccagaaatac accaacagca
     7741 cgcatgatgt ctactgggga tttattcttt agtgcggggg aatacacggc ttttaatacg
     7801 attgagggcg tctcctaaca agttacatca ctcctgccct tcctcaccct catctccatc
     7861 acctccttca tctccgtcat ctccgtcatc accctccgcg gcagcccctt ccaccatagg
     7921 tggaaaccag ggaggcaaat ctactccatc gtcaaagctg cacacagtca ccctgatatt
     7981 gcaggtagga gcgggctttg tcataacaag gtccttaatc gcatccttca aaacctcagc
     8041 aaatatatga gtttgtaaaa agaccatgaa ataacagaca atggactccc ttagcgggcc
     8101 aggttgtggg ccgggtccag gggccattcc aaaggggaga cgactcaatg gtgtaagacg
     8161 acattgtgga atagcaaggg cagttcctcg ccttaggttg taaagggagg tcttactacc
     8221 tccatatacg aacacaccgg cgacccaagt tccttcgtcg gtagtccttt ctacgtgact
     8281 cctagccagg agagctctta aaccttctgc aatgttctca aatttcgggt tggaacctcc
     8341 ttgaccacga tgctttccaa accaccctcc ttttttgcgc ctgcctccat caccctgacc
     8401 ccggggtcca gtgcttgggc cttctcctgg gtcatctgcg gggccctgct ctatcgctcc
     8461 cgggggcacg tcaggctcac catctgggcc accttcttgg tggtattcaa aataatcggc
     8521 ttcccctaca gggtggaaaa atggccttct acctggaggg ggcctgcgcg gtggagaccc
     8581 ggatgatgat gactgactac tgggactcct gggcctcttt tctccacgtc cacgacctct
     8641 ccccctggct ctttcacgac ttccccccct ggctctttca cgtcctctac cccggcggcc
     8701 tccactacct cctcgacccc ggcctccact acctcctcga ccccggcctc cactgcctcc
     8761 tcgaccccgg cctccacctc ctgctcctgc ccctcctgct cctgcccctc ctcctgctcc
     8821 tgcccctcct gcccctcctg ctcct
//
