LOCUS       sequence-275051-        8489 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 8489)
  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 8489)
  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: pHM151Env
FEATURES             Location/Qualifiers
     source          1..8489
                     /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"
     misc_feature    2769..3002
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3525..3551
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3987..4028
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4038..4055
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4078..4095)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4084..4308
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4354..4782
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4441..4460)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4651..4672
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4791..4811)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4796..5125
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4976..5111
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5038..5057
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5192..5986
                     /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(5246..5265)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5856..5875
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6162..6283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6199..6218)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6253..6272
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6332..6348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6332..6348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6345..6367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6356..6372
                     /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(6380..6386,6387..6404,6405..6410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6425..6446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6563..6580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6734..7322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6814..6833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7493..8284,8285..8353))
                     /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     8116..8135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8354..8458)
                     /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 gaagcggaga cagcgacgaa gagctcctca agacagtaag actcatcaag
     1021 cttcttcatc aaagcagtaa gtagtatatg taatgcaagc tttagagata tatgcaatag
     1081 tagcattagt aatagtagga ataattgcaa tagttgtgtg gactatagta cttatagaat
     1141 ataagaaaat attaagacaa agaagaatag acaggttact tgaaagaata agagaaagag
     1201 cagaagacag tggcaatgag agtgaggggg atcaggaaga attacttatg gaaatggggg
     1261 aaatggggca tcttattcct ggggatattg atgatctgta gcgctgcaga caatttgtgg
     1321 gtcacagtct attatggggt acctgtgtgg agagatgcaa acaccactct attttgtgca
     1381 tcagatgcta aagcatatga aagagaagga cataatgttt gggccacaca tgcatgtgta
     1441 cccacagacc ccaacccaca agaattggta ttggaaaatg tgacagaaga gtttaatatg
     1501 tggaaaaata acatggtaga acagatgcat gaggacataa tcagtttatg ggatcaaagc
     1561 ctaaaaccat gtgtaaaaat aaccccactc tgtgtcacat taaactgcac taacgctacc
     1621 aataacgcta ccaatagtaa taataccagt aataaaataa agggggaggt agtaatgaaa
     1681 aactgctctt tcaatgtcac cacagacata aaagataaga gaatagaaag agcattgttt
     1741 tataaagatg atttagtaca aataggggag aataatacta taggggagaa taatactata
     1801 tataggttaa taagctgtaa ctcctcagtc attacccaag cctgtccaaa ggtatccttt
     1861 gagccaattc ccatacatta ttgtaccccg gctggttatg cgattcttaa gtgtaataat
     1921 aagacattta atggatcagg gctatgtaca aatgtcagca cagtacaatg tacacatgga
     1981 attaggccag tagtatcaac tcaactgctg ttaaatggca gtttagcaga agaagaggta
     2041 gtaattagat ctgaaaattt cacaaacaat gctaaaacca taatagtaca gctgaaagaa
     2101 gctgtagtaa ttaattgcac aagacccaac aacaatacaa gaaaaagtat acatataggg
     2161 caaggaagag catggtatac aacaggagag gtagtgggag acataagaca agcatattgt
     2221 acaattaaca aaacaaaatg gaataacact ttaaaccaga tagttgaaaa attaagaggt
     2281 caatttggga ataaaacaat aatctttact cagtcctcag gaggggaccc agagattgta
     2341 atgcacagtt ttaattgtgg aggggaattt ttctactgta atacatcaca actatttaat
     2401 agtacttgga atggaaatag tagtacttgg gatggaaata gtactaggga gaatagcacc
     2461 acaatcacac tcccgtgcag aataagacaa attataaaca tgtggcagaa agtaggaaaa
     2521 gcaatgtatg cccctcccat cgcgggtcca attatctgtt tatcaaacat tacagggctg
     2581 ttattaacaa gagatggtgg tggtaataat acaattaatg agaccttcag accaggagga
     2641 ggagatatga gggacaattg gagaagtgaa ctatacaaat ataaagtagt aagaattgaa
     2701 ccattaggac tggcaccctc caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     2761 gtgggactag gagcgttgtt cattgggttc ttgggagcag caggaagcac tatgggcgca
     2821 gcgtcagtga cgctgacggt acaggccaga caattattgt ctggtatagt gcaacagcag
     2881 aacaatctgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     2941 atcaagcagc tccaggcaag agtcctggct gtggaaagat acctagggga tcaacagctc
     3001 ctaggaatgt ggggttgctc tggaaaactc atttgcccca cctctgtgcc ttggaatgct
     3061 agttggagca ataaatctct ggaagagatt tggaataaca tgacctggag gcaatgggag
     3121 ggagaaattg acaattacac aggcttaata tacaccttac ttacagaatc gcagagccaa
     3181 caggaaaaga atgaacaaga attattagaa ttggataaat gggcaaattt atggacttgg
     3241 tttgacatat caaactggct gtggtatata aaaatattca taatgatagt aggaggcttg
     3301 ataggcttaa gaatagtttt tactgtactt tctataataa atagagttag gcagggatac
     3361 tcaccattgt cgttgcagac ccacctccca gctcagaggg gacccgacag gccagaagga
     3421 atcgaagaag aaggtggaga gagagacaga gacggatccg gacgattagt gggtggattc
     3481 ttgacactta tctgggtcga cctgaggagc ctgttcctct acagctacca ccgcttgaga
     3541 gacttactct tgattgtaac gaggattgtg gaacttctgg gacgcagggg gtgggaactc
     3601 ctcaagtact ggtggaatct cctacagtat tggagtcagg aactaaagaa tagtgctact
     3661 agcttgctca acaccacagc catagcagta gctgagggga cagataggct tatagaatta
     3721 gtgcaattag cttttagagc tattctccac atacctagaa gaataagaca gggcttagaa
     3781 agggctttgc tataagatgg gtggcaagtg gtcaaaacgc agtaggagtg aatggcctgc
     3841 tgtaagggaa agaatgcaac aagttgagcc agcagcagaa ggggtgggag cagtatctcg
     3901 agacggatcc agctaagggt caagacaatt ctgcagatat ccagcacagt ggcggccgct
     3961 cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     4021 attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct
     4081 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4141 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4201 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4261 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     4321 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4381 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4441 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4501 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4561 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4621 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4681 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4741 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4801 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4861 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4921 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4981 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5041 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     5101 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     5161 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5221 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5281 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5341 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5401 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5461 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5521 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5581 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5641 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5701 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5761 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5821 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5881 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5941 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     6001 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6061 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6121 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6181 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6241 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6301 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6361 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6421 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6481 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6541 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6601 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6901 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7141 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7381 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7441 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7501 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7561 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7621 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7681 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7741 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7801 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7861 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7921 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7981 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8041 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8101 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8161 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8221 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8281 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8341 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8401 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8461 catttccccg aaaagtgcca cctgacgtc
//
