LOCUS       sequence-267120-        8492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Env gene from HIV-1 LAI in pcDNA3.1 expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8492)
  AUTHORS   Joshi A, Sedano M, Beauchamp B, Punke EB, Mulla ZD, Meza A, Alozie 
            OK, Mukherjee D, Garg H
  TITLE     HIV-1 Env Glycoprotein Phenotype along with Immune Activation 
            Determines CD4 T Cell Loss in HIV Patients.
  JOURNAL   J Immunol. 2016 Feb 15;196(4):1768-79. doi: 
            10.4049/jimmunol.1501588. Epub 2016 Jan 13.
  PUBMED    26764036
REFERENCE   2  (bases 1 to 8492)
  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"; doi: 
            "10.4049/jimmunol.1501588"; journalName: "J Immunol"; date: 
            "2016-02-15- 15"; volume: "196"; issue: "4"; pages: "1768-79"
COMMENT     Sequence Label: pLAI-Env
FEATURES             Location/Qualifiers
     source          1..8492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        172..551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        552..755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     706..726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     800..819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        800..818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2709..2942
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3127..3171
                     /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             3465..3491
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3927..3968
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3978..3995
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4018..4035)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4024..4248
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4294..4722
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4381..4400)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4591..4612
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4731..4751)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4736..5065
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4916..5051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4978..4997
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5132..5926
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5186..5205)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5796..5815
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6102..6223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6139..6158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6193..6212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6272..6288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6272..6288)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6285..6307)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6296..6312
                     /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)."
     promoter        complement(join(6320..6326,6327..6344,6345..6350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6365..6386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6503..6520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6674..7262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6754..6773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7433..8224,8225..8293))
                     /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     8056..8075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8294..8398)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8473..8492)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
       61 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
      121 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      181 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      541 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      601 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      661 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      721 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      781 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagttaa
      841 gcttggtacc gagctcggat ccagtaccct tcaccgatca agctttaggc atctcctatg
      901 gcaggaagaa gcggagacag cgacgaagac ctcctcaagg cagtcagact catcaagttt
      961 ctctatcaaa gcagtaagta gtacatgtaa tgcaacctat acaaatagca atagcagcat
     1021 tagtagtagc aataataata gcaatagttg tgtggtccat agtaatcata gaatatagga
     1081 aaatattaag acaaagaaaa atagacaggt taattgatag actaatagaa agagcagaag
     1141 acagtggcaa tgagagtgaa ggagaaatat cagcacttgt ggagatgggg gtggaaatgg
     1201 ggcaccatgc tccttgggat attgatgatc tgtagtgcta cagaaaaatt gtgggtcaca
     1261 gtctattatg gggtacctgt gtggaaggaa gcaaccacca ctctattttg tgcatcagat
     1321 gctaaagcat atgatacaga ggtacataat gtttgggcca cacatgcctg tgtacccaca
     1381 gaccccaacc cacaagaagt agtattggta aatgtgacag aaaattttaa catgtggaaa
     1441 aatgacatgg tagaacagat gcatgaggat ataatcagtt tatgggatca aagcctaaag
     1501 ccatgtgtaa aattaacccc actctgtgtt agtttaaagt gcactgattt ggggaatgct
     1561 actaatacca atagtagtaa taccaatagt agtagcgggg aaatgatgat ggagaaagga
     1621 gagataaaaa actgctcttt caatatcagc acaagcataa gaggtaaggt gcagaaagaa
     1681 tatgcatttt tttataaact tgatataata ccaatagata atgatactac cagctatacg
     1741 ttgacaagtt gtaacacctc agtcattaca caggcctgtc caaaggtatc ctttgagcca
     1801 attcccatac attattgtgc cccggctggt tttgcgattc taaaatgtaa taataagacg
     1861 ttcaatggaa caggaccatg tacaaatgtc agcacagtac aatgtacaca tggaattagg
     1921 ccagtagtat caactcaact gctgttgaat ggcagtctag cagaagaaga ggtagtaatt
     1981 agatctgcca atttcacaga caatgctaaa accataatag tacagctgaa ccaatctgta
     2041 gaaattaatt gtacaagacc caacaacaat acaagaaaaa gtatccgtat ccagagggga
     2101 ccagggagag catttgttac aataggaaaa ataggaaata tgagacaagc acattgtaac
     2161 attagtagag caaaatggaa tgccacttta aaacagatag ctagcaaatt aagagaacaa
     2221 tttggaaata ataaaacaat aatctttaag caatcctcag gaggggaccc agaaattgta
     2281 acgcacagtt ttaattgtgg aggggaattt ttctactgta attcaacaca actgtttaat
     2341 agtacttggt ttaatagtac ttggagtact gaagggtcaa ataacactga aggaagtgac
     2401 acaatcacac tcccatgcag aataaaacaa tttataaaca tgtggcagga agtaggaaaa
     2461 gcaatgtatg cccctcccat cagcggacaa attagatgtt catcaaatat tacagggctg
     2521 ctattaacaa gagatggtgg taataacaac aatgggtccg agatcttcag acctggagga
     2581 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     2641 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     2701 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     2761 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     2821 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     2881 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     2941 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     3001 agttggagta ataaatctct ggaacagatt tggaataaca tgacctggat ggagtgggac
     3061 agagaaatta acaattacac aagcttaata cattccttaa ttgaagaatc gcaaaaccag
     3121 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     3181 tttaacataa caaattggct gtggtatata aaaatattca taatgatagt aggaggcttg
     3241 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     3301 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     3361 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcc
     3421 ttagcactta tctgggacga tctgcggagc ctgtgcctct tcagctacca ccgcttgaga
     3481 gacttactct tgattgtaac gaggattgtg gaacttctgg gacgcagggg gtgggaagcc
     3541 ctcaaatatt ggtggaatct cctacagtat tggagtcagg aactaaagaa tagtgctgtt
     3601 agcttgctca atgccacagc catagcagta gctgagggga cagatagggt tatagaagta
     3661 gtacaaggag cttgtagagc tattcgccac atacctagaa gaataagaca gggcttggaa
     3721 aggattttgc tataagatgg gtggcaagtg gtcaaaaagt agtgtggttg gatggcctac
     3781 tgtaagggaa agaatgagac gagctgagcc agcagcagat ggggtgggag cagtatctcg
     3841 agacggatcc agctaagggt caagacaatt ctgcagatat ccagcacagt ggcggccgct
     3901 cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     3961 attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct
     4021 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4081 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4141 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4201 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     4261 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4321 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4381 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4441 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4501 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4561 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4621 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4681 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4741 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4801 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4861 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4921 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4981 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     5041 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     5101 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5161 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5221 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5281 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5341 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5401 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5461 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5521 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5581 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5641 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5701 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5761 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5821 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5881 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5941 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6001 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6061 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6121 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6181 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6241 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6301 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6361 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6421 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6481 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6541 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6601 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6661 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6721 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6781 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6841 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6901 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6961 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7021 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7081 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7141 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7261 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7321 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7381 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7441 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7501 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7561 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7621 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7681 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7741 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7801 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7861 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7921 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7981 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8041 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8101 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8161 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8221 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8281 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8341 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8401 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     8461 gtgcactctc agtacaatct gctctgatgc cg
//
