LOCUS       sequence-273365-        8499 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 8499)
  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 8499)
  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: pHM123DEnv
FEATURES             Location/Qualifiers
     source          1..8499
                     /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             3535..3561
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3997..4038
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4048..4065
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4088..4105)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4094..4318
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4364..4792
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4451..4470)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4661..4682
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4801..4821)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4806..5135
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4986..5121
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5048..5067
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5202..5996
                     /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(5256..5275)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5866..5885
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6172..6293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6209..6228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6263..6282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6342..6358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6342..6358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6355..6377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6366..6382
                     /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(6390..6396,6397..6414,6415..6420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6435..6456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6573..6590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6744..7332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6824..6843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7503..8294,8295..8363))
                     /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     8126..8145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8364..8468)
                     /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 gaagcggaag caccgacgaa gaactcctca gagcaataag gatcatcaaa
     1021 atcctatacc agagcagtaa gtaataaata tatgtaatga cacctttaac acctttgcaa
     1081 attagtgcaa tagcaggact gatagtagcg ctaatcttag caatagtagt gtggactata
     1141 gtagctatag aatttaagaa aatattaagg gaaagaaaga taaaaaggtt agttaagaga
     1201 ataagagaaa gagcagaaga cagtggaaat gaaagtgaag gggacacaga tgaattggcc
     1261 aaacttgtgg agatggggga ctttgatcct tgggttggta ataatgtgta gtgcctcaga
     1321 caacttgtgg gttacagttt attatggggt tcctgtgtgg aaagatgcag ataccacctt
     1381 attttgtgca tcagatgcca gagtacatga cacagaagtg cacaatgtct gggccacaca
     1441 tgcctgtgta cccacagacc ccaacccaca agaaatacac ctggaaaatg taacagaaaa
     1501 ttttaatatg tggaaaaaca acatggtaga gcagatgcat gaggatgtaa tcagtctatg
     1561 ggatcaaagt ctaaagccat gtgtaaagtt aactcctctc tgtgttactt taaattgtac
     1621 caatactacc aatgctaatt tgacccaggg caaaatcaca accactagcc ctaacatagg
     1681 aaatacaaca gatgaagtaa aatgttcttt taatataacc acagtactaa aagataagaa
     1741 gcagcaggtc actgcacttt tttataagct tgatgtagtg caaattgatg aggagggtaa
     1801 taagagtaaa agtagtagta atagtggtaa gtataggtta ataaattgta atacttcagt
     1861 cattaaacag gcttgtccaa agatatcttt tgatccaatt cctatacatt attgtactcc
     1921 agctggttat gcgcttttaa agtgtaatga taaaaatttc aatgggacag ggccatgtaa
     1981 taatgtcagt tcagtacaat gcacacatgg aattaagcca gtggtatcaa ctcaattgct
     2041 gttaaatggc agtctagcag aagaaaaaat aataatcagg tctgaaaata tcacagataa
     2101 taccaaaatc ataatagtgc accttaatac atctgtaaaa atcaattgta ccagacctgc
     2161 caacaataca agaacaagta tagctatagg accaggacaa atgttcttct atagacaagg
     2221 agatgtaata ggaaatataa gaaaggcaca gtgtgaggtt aatggagcca aatggaatgc
     2281 agttttaaaa caggtaaaaa gaacattaga aaagcagatt aataagacaa taaactttaa
     2341 acaaccatcc tcaggaggag atctagaaat tacaatgcat cattttaatt gtagagggga
     2401 atttttctat tgcaatacaa caccactgtt taataacact tgcctagaca ccgggaattg
     2461 tactggcagt atcacacttc catgcaagat aaagcaaatt ataaacatgt ggcagggagt
     2521 aggacaagca atgtatgctc ctcctatcag tggaagcatt aattgtgtat cagatatcac
     2581 aggactacta ttgacaagag atggtggtgg taatgagaca agcaagaacg agaccttcag
     2641 acctgcagga ggaaatatga aggacaattg gagaagtgaa ttatataaat ataaagtagt
     2701 acaaattgaa ccactaggaa tagcacccac cagggcaaag agaagagtgg tggagagaga
     2761 aaaaagagca gtgggaatag gagctatgat ctttgggttc ttaggagcag caggaagcac
     2821 tatgggcgcg gcgtcaataa cgctgacggt acaggccaga caattattgt ctggtatagt
     2881 gcaacagcaa agcaatttgc tgagggctat agaggcgcag cagcatatgt tgcaactcac
     2941 agtctggggc attaaacagc tccaggcaag agtcctggct gtggaaagat acctaaaaga
     3001 tcaaaagttc ctaggacttt ggggctgctc tggaaaaatc atctgcacca ctgctgtgcc
     3061 ctggaactcc acttggagta ataaatctta tgaagaaatt tggaacaaca tgacatgggc
     3121 acaatgggag aaagaaatta gcaattacac agatcaaata tatgacatac ttacagaatc
     3181 gcagaaccag caggacagga atgaaaagga tttgttagaa ttggataaat gggcaagcct
     3241 gtggaattgg tttgacataa caagttggct gtggtatata aaaatattta taatgatagt
     3301 aggaggttta atagggttaa gaataatttt tgctgtgctt tctatagtaa atagagttag
     3361 gcagggatac tcacctttgt ctttccagat ccctacccat caacagaggg aacccgacag
     3421 gcccgaagga atcgaagaag gaggtggcga gcaaggcaga gacagatccg tgagattagt
     3481 gaccggattc ttagcacttg cctgggacga cctgcggagc ctgtgcctct tctgctaccg
     3541 ccgcttgaga gacttactct tgattgcagc gaggactgtg gaacttctgg gacgcagggg
     3601 gagggaaggc ctcaaatatc tggggaatct tctgttgtat tggggtcagg agctaaaaat
     3661 tagtgccatt tctttgctta atgccacagc aatagcagta gcggggtgga cagatagggt
     3721 tatagaagta gcacaaggag cttggagagc cattctccac atacctagaa gaatcagaca
     3781 gggcttagaa agggctttgc tataacatgg gaaacaagtg gtcaaaaagt agcatagtgg
     3841 gatggcctca aatcagggaa agaataagac gaactcctcc agcagcagaa ggagtaggag
     3901 cagtatctca agacggatcc agctaagggt caagacaatt ctgcagatat ccagcacagt
     3961 ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
     4021 ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc
     4081 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     4141 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4201 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4261 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     4321 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4381 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4441 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4501 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4561 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4621 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4681 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4741 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4801 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4861 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4921 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4981 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5041 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     5101 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5161 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5221 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5281 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5341 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5401 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5461 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5521 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5581 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5641 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5701 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5761 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5821 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5881 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5941 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6001 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6061 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6121 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6181 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6241 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6301 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6361 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6421 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6481 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6541 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6601 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6661 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6721 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6781 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6841 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6901 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6961 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7021 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7081 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7141 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7201 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7261 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7321 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7381 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7441 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7501 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7561 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7621 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7681 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7741 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7801 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7861 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7921 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7981 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8041 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8101 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8161 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8221 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8281 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8341 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8401 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8461 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
