LOCUS       sequence-296768-        8475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HIV Env which is isolated a from IRB approved HIV infected 
            patient samples from Singapore and cloned into expression vector 
            (pcDNA 3.1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8475)
  AUTHORS   Joshi A, Cox EK, Sedano MJ, Punke EB, Lee RT, Maurer-Stroh S, Kaur 
            P, Ng OT, Garg H
  TITLE     HIV-1 subtype CRF01_AE and B differ in utilization of low levels of 
            CCR5, Maraviroc susceptibility and potential N-glycosylation sites.
  JOURNAL   Virology. 2017 Dec;512:222-233. doi: 10.1016/j.virol.2017.09.026. 
            Epub 2017 Oct 9.
  PUBMED    29020646
REFERENCE   2  (bases 1 to 8475)
  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: "Virology.";
            date: "2017-12"; volume: "512:222-233. doi"; pages: " 
            10.1016/j.virol.2017.09.026. Epub 2017 Oct 9"
COMMENT     Sequence Label: pHM080Env
FEATURES             Location/Qualifiers
     source          1..8475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3181..3225
                     /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             3519..3545
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3973..4014
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4024..4041
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4064..4081)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4070..4294
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4340..4768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4427..4446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4637..4658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4777..4797)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4782..5111
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4962..5097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5024..5043
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5178..5972
                     /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(5232..5251)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5842..5861
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6148..6269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6185..6204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6239..6258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6318..6334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6318..6334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6331..6353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6342..6358
                     /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(6366..6372,6373..6390,6391..6396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6411..6432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6549..6566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6720..7308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6800..6819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7479..8270,8271..8339))
                     /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     8102..8121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8340..8444)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccagtac ccttcaccga tcaagcttta ggcatctcct
      961 atggcaggaa gaagaggaga cagcgacgaa gacctcctca acacagtcag aatcatcaag
     1021 ctcctgtacc agagcagtaa gtaatacatg acatgcaaac tttaacaata ttaacaatag
     1081 tagcattagt agtagcagca atactagcaa tagttgtgtg gactatagtg ttcatagaat
     1141 ataggaaaat aaaaagacaa aagaaaatag ataggttaat tgatagaata agagaaagag
     1201 cagaagacag tggcaatgaa agtgaagggg atcaggaaga attgtcaaaa cttgtggaaa
     1261 tggggcacct tgctccttgg gatattgatg atttgtagtg cagaagacaa ttggtgggtc
     1321 acagtctatt atggagtacc tgtatggaaa aatgcaacca ccactctatt ttgtgcatca
     1381 gatgctaaag catatgatac agaggtacac aatgtttggg ccacacatgc ctgtgtaccc
     1441 acagacccca acccacaaga agtagtattg ggaaatgtga cagaatattt taacatgtgg
     1501 gaaaatgata tggtagaaca gatgcaggag gatgtaatca gtttatggga tcaaagctta
     1561 aagccatgtg tgaaattaac cccactctgt gtcactttaa attgcactga gttaaatact
     1621 actagtaatg ctactgggtt aaatactact agtaatacta gcactgtgtc acataatagt
     1681 aaggatgaaa tgaaaaactg ctctttcaat gtcacctcaa acataagaga taagttgcag
     1741 agagaatatg cacttttcta taaacttgat atagtaccaa tagataatga taatagcagc
     1801 tatagattga taagttgtaa cacctcagtc attacacagg cgtgtccaaa ggtatctttt
     1861 acgccaattc ctatacatta ttgtactcca gctggttttg cgcttctaaa gtgtaacaat
     1921 aagacattca atggaaaagg accatgtaca aatgtcagca ctgtacagtg tactcatggg
     1981 attaggccag tagtatcaac tcagctgttg ttaaatggca gtctagcaga agaagaagta
     2041 ataattagag ctgacaattt cactgataac gccaaaacca taatagtaca gctgaaggaa
     2101 cctgtaaaaa ttaattgtac aagaccccac aacaatacaa gaaagagtat acatatagga
     2161 ccagggagag cattctatgc aacaacagat ataataggag atataagaca agcacattgt
     2221 aacattagtg caaaagactg gggaaaaact ttacaacagg tagctgaaaa attaaaagaa
     2281 caatttaaga acaaaacaat atactttaat caatcctcag gaggggaccc agaaattgta
     2341 atgcatagtg ttaattgtgg aggggaattt ttttactgta atacatcaaa actgtttaat
     2401 agtacttggc tgaataataa tacagggtca aatgccactg aacctatcat actcccatgc
     2461 aggataaaac aaattgtaaa catgtggcag gaagtaggaa aagcaatgta tgcccctccc
     2521 atcagaggac aaattagatg catatcaaat attacagggc tcctattaac aagagatggt
     2581 ggtcaagaga acaaaaccga gaaccacacc gaggtcttta gacccacggg aggaaatatg
     2641 aaggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     2701 atagcaccca ccaaggcaaa aagaagagtg gtgcagagag aaaaaagagc agtgggaata
     2761 ggagcgatgt tccttgggtt cttgagcgca gcaggaagca ctatgggcgc agcgtcaata
     2821 acgctgacgg tacaggccag acagttactg tctggtatag tgcaacagca aaacaatttg
     2881 ctgagagcta ttgaggcgca acaacatatg ttgcaactca cagtctgggg catcaagcag
     2941 ctccaggcaa gagtcctggc tatagaaaga tacctaaggg atcaacagct cctggggatt
     3001 tggggctgct ctggaaaact catttgcacc actaatgtgc cttggaatgc tagttggagt
     3061 aaaaataaaa gtatagatga ggtttggaat aacatgacct ggatgcagtg ggaaaatgag
     3121 attgacaatt acacaggctt aatatatact ttacttgaag aatcgcaaaa ccagcaagaa
     3181 aagaatgagc aagaattatt ggaattggat aaatgggcaa gtttgtggaa ttggtttgac
     3241 ataacaaatt ggctgtggta tataaaaata ttcataatga tagtaggagg cttgataggc
     3301 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggaaggg atactcacca
     3361 ttgtcattcc agacccacct cccagctccg aggggactcg acaggcccga aggaatcgac
     3421 gaagaaggtg gagagagaga cagagacaca tccgggccat tagtgaacgg attcttagcg
     3481 atcatctggg tcgacctacg gagcctgtgc ctcttcagct accaccgctt gagagactta
     3541 ctcttgattg tagcgaggat tgtggaactt ctgggacgca gggagtggga agccctcaag
     3601 tattggtgga gtctcctaca gtattggagt caggaactaa agaatagtgc tgttagtttg
     3661 cttaatacca tagctatagc tgtagctgag ggaacagata gggttataga aatagtacaa
     3721 agagtttgta gagctattcg caacatacct agaagaataa gacagggctt agaaagggct
     3781 ttgcagtaag atgggtggca agtggtcaaa acgtagtggg ggtggatggt ttacagtaag
     3841 agaaaaaatg agagaagctc cagcagcaga cggggtggga gcagtatctc gagacggatc
     3901 aagggtcaag acaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc
     3961 ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc
     4021 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct
     4081 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     4141 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     4201 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     4261 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     4321 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4381 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4441 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4501 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4561 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4621 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4681 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4741 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4801 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4861 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4921 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4981 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5041 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5101 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     5161 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5221 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5281 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5341 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5401 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5461 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5521 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5581 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5641 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5701 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5761 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5821 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5881 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5941 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc
     6001 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     6061 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6121 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6181 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6241 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6301 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6481 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7141 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7381 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7441 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7501 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7561 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7621 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7681 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7741 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7801 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7861 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7921 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7981 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8041 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8101 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8161 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8221 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8281 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8341 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8401 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8461 gtgccacctg acgtc
//
