LOCUS       sequence-273349-        8491 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 8491)
  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 8491)
  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: pHM032Env
FEATURES             Location/Qualifiers
     source          1..8491
                     /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             3989..4030
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4040..4057
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4080..4097)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4086..4310
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4356..4784
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4443..4462)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4653..4674
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4793..4813)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4798..5127
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4978..5113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5040..5059
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5194..5988
                     /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(5248..5267)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5858..5877
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6164..6285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6201..6220)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6255..6274
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6334..6350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6334..6350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6347..6369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6358..6374
                     /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(6382..6388,6389..6406,6407..6412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6427..6448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6565..6582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6736..7324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6816..6835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7495..8286,8287..8355))
                     /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     8118..8137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8356..8460)
                     /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 caccgacgag gaactcctca gagcagtaag gatcatcaga
     1021 atcctatacc agagcagtaa gtaataagta tctgttatgt cacctttgga aattagtgca
     1081 atagcaggac tgataatagc gctaatcgca gcaatagtag tgtggactat agtagctata
     1141 gaatttagga aaatattaaa gcaaagaaaa atagacaggt tagttaagag gataagagaa
     1201 agagcagaag acagtggaaa tgagagtgaa ggagacacag atgaattggc caaactggtg
     1261 gaaatggggg actttgatcc ttgggttggt gataatttgt agtgcctcaa acaacttgtg
     1321 ggttacagtt tattatgggg ttcctgtgtg gagagatgcc gataccacct tattttgtgc
     1381 atcaaatgcc aaagcacatg agacagaagt gcacaatgtc tgggccacac atgcctgtgt
     1441 gcccacagat cccaacccac aagaaataca cctggaaaat gtaacagaaa attttaacat
     1501 gtggaaaaat aaaatggcag agcagatgca ggaggatgta atcagtttat gggatcaaag
     1561 tctaaagcca tgtgtaaagt taactcctct ctgcgttact ttaacttgta ccaatgctag
     1621 tttgaccaat gttactttga ccagggtcaa taacaccaat gactatagca tagtaggaaa
     1681 tataacagat gaaataagaa actgttcttt taatatgacc acagaaataa atgataagaa
     1741 gcagaaggtc catgcacttt tttataggct tgatatagta ccaatgggaa atgaaactgg
     1801 gggtgtgggt gaatataggt taataaattg taatacttca gtcattaagc aggcttgtcc
     1861 aaagataact tttgatccaa ttcctataca ttattgcact ccagctggtt atgcgatttt
     1921 aaaatgtaat gataaggatt tcaatgggac agggccatgt aaaaatgtca gctcagtaca
     1981 atgcacacat ggaattaagc cagtggtatc aactcaattg ctgttaaatg gcagtctagc
     2041 agaaggagag ataataatca gatctgaaaa tctcacaaac aatgccaaaa acataatagt
     2101 gcaccttaat aagagtgtag aaatcaattg taccagaccc tccaacaata caagaacaag
     2161 tataactata ggaccaggaa gagtattcta taaaacagga gacataatag gagatataag
     2221 acaagcatat tgtgagatta atggaacaaa atggaataaa gtattaaaac aagtagctga
     2281 gaaattaaaa gagcacatta ataagacaat aagctttaaa aaaccctcag gaggagatct
     2341 agaaattaca atgcatcatt ttaattgtag aggggaattt ttctattgca atacaacaaa
     2401 actgtttaat aacacttgca taaatggaac aatggaaaat gagagtaata gcactatcat
     2461 acttccatgc aagattaaac aaattataaa catgtggcag ggagcaggac aagcaatgta
     2521 tgctcctccc atcagtggaa gaattaattg tatatcaaat attacaggaa tactattaac
     2581 aagagatggt cagccgaatg atacggagat cttcagacct ggaggaggag atatgaagga
     2641 caattggaga aatgaattat ataaatataa agtagtacaa attgaaccac taggaatagc
     2701 acccaccagg gcaaagagaa gagtggtgga gagagaaaaa agagcagtgg gaataggagc
     2761 gatgatcttt gggttcttag gagcagcagg aagcactatg ggcgcggcgt caataacgct
     2821 gacggtacag gccagacagt tattgtctgg tatagtgcaa cagcaaagca atttgctgag
     2881 ggctatagag gcgcagcagc atctgctgca actcacagtc tggggcatta aacagctcca
     2941 ggcaagagtc ctggctgtgg aaagatacct aaaagatcaa aagttcctag gactttgggg
     3001 ctgctctgga aaaatcatct gcaccactgc tgtaccctgg aactcctctt ggagtaatag
     3061 atcttatgaa gagatttgga acaacatgac atggatagaa tgggagagag aaattagcaa
     3121 ttacacaaac caaatatatg agatacttac agaatcgcag aaccagcagg acagaaatga
     3181 aaaggatttg ttagaattgg ataaatgggc aagtctgtgg agttggttcg acataacaag
     3241 atggctgtgg tatataagaa tatttataat aatagtagga agtctaatag gtttaagaat
     3301 aatttttgct gtgctttcta tagtaaatag agttaggcag ggatactcac ctttatcttt
     3361 ccagaccctt acccatcatc agagggaacc cgacaggccc gaaagaatcg aagaaggcgg
     3421 tggcgagcaa ggcagagaca gatccgtgag atttgtgagc ggattcttag cgcttgcgtg
     3481 ggacgacctg cggagcctgt gcctcttttg ctaccaccgc ttgagagact tcatcttgat
     3541 tgcaacgagg actgtggaac ttctgggaca cagcagtctc aagggactga gacgggggtg
     3601 ggaaggcctc aaatatctgg ggaaccttct gttatattgg ggccaggaac taaaaattag
     3661 tgctatctct ttgtttaatg ctacagcaat aacagtagcg gggtggacag atagggttat
     3721 cgaagtcgca caaagagctt ggagagcctt tctccacata cctagaagaa tcagacaggg
     3781 cttagaaagg gctttgcaat aacatgggaa gcaagtggtc aaaaagtagc atagtgggat
     3841 ggcctcaggt cagagaaaga ataaggcaaa ctcctccagc agcagaagga gtaggagcag
     3901 tatctcaaga cggatcaagg gtcaagacaa ttctgcagat atccagcaca gtggcggccg
     3961 ctcgagtcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc tcctcggtct
     4021 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     4081 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4141 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4201 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4261 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4321 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4381 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4441 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4501 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4561 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4621 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4681 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4741 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4801 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4861 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4921 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4981 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5041 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5101 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5161 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5221 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5281 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5341 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5401 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5461 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5521 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5581 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5641 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5701 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5761 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5821 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5881 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5941 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6001 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6061 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6121 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6181 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6241 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6301 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6361 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6481 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6541 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6601 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7141 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7561 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7621 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7681 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7741 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7801 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7861 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7921 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7981 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8041 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8101 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8161 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8221 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8281 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8341 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8401 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8461 cacatttccc cgaaaagtgc cacctgacgt c
//
