LOCUS       sequence-274205-        8458 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 8458)
  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 8458)
  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: pHM040Env
FEATURES             Location/Qualifiers
     source          1..8458
                     /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             3956..3997
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4007..4024
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4047..4064)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4053..4277
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4323..4751
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4410..4429)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4620..4641
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4760..4780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4765..5094
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4945..5080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5007..5026
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5161..5955
                     /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(5215..5234)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5825..5844
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6131..6252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6168..6187)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6222..6241
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6301..6317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6301..6317)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6314..6336)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6325..6341
                     /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(6349..6355,6356..6373,6374..6379))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6394..6415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6532..6549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6703..7291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6783..6802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7462..8253,8254..8322))
                     /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     8085..8104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8323..8427)
                     /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 tcagctttag gcatctccta
      961 tggcaggaag aagcggaagc accgacgagg aactcctcgc agcagtaaga atcatcaaga
     1021 tcttatacca gagcagtaag tatatgtaat gtcaccttta gaaattagta caatagtagg
     1081 actgatagta gcattaatct tagcaatagt agtgtggact atagtagcta tagaacttaa
     1141 gaaactacta agagaaagaa aaatagaaag attagttaaa agaataagag agagagcaga
     1201 agacagtgga aatgagagtg agggggacac ggatgaattg gcaaaacttg tggaaatggg
     1261 ggactttgat ccttgggttg gggataattt gtagtgcctc agacaacttg tgggttacag
     1321 tttattatgg ggttcctgtg tggagagatg cagacaccac cctattttgt gcatcagatg
     1381 cccaagcaca taagacagaa gtgcacaatg tctgggccac acatgcctgt gtacccacag
     1441 accccaaccc acaagaaata ctcctggaaa atgtaacaga aaattttaac atgtggaaaa
     1501 ataacatggt ggagcagatg caggaagatg taatcagttt atgggatcaa agtctaaagc
     1561 catgtgtaaa gttaactcct ctctgcgtta ctttaaattg taccaatgct aatctgactg
     1621 ccacgaactc aaacaatgtc actaacatag taggaaaggt aacagaggaa gtaaaaaact
     1681 gctcttttaa tatgacaaca gaactaacag ataggaggca gaaggtccat gcactttttt
     1741 ataaacttga tatagtacaa atggatacaa aaaatagtag tgacaattat agtgagtata
     1801 ggataataaa ctgtaatact tcagtcatta agcaggcttg tccaaaggta tcctttgatc
     1861 caattcctat acattattgt actccagctg gttatgcgat tttaaaatgt aatgataaga
     1921 agttcaatgg aacagggcca tgtagaaatg tcagctcagt acaatgcaca catgggatta
     1981 agccagtggt atcaactcag ttgctgttaa atggtagtct agcagaagat gagatagtaa
     2041 tcagaagtga aaatctcaca gacaatgcca agaccataat agtgcagctt aataagtctg
     2101 tagacatcaa ttgtaccaga ccctccaaca atacaagaac aagtgtgtct ataggaccag
     2161 gacaagtatt ctatagaaca ggagacataa taggagatat aacaaaagca tattgtgaga
     2221 ttaatggaac aaaatggaat gaaactttaa cacaggtagc taaaaaatta aaggagcact
     2281 ttaataagac aataatcttt caaccacact caggaggaga tctagaaatt acaatgcatc
     2341 attttaattg tagaggggaa tttttctatt gcaatacaac aaaactgttt aatggtactt
     2401 gcccaaaaag tggaacatgt aatggcacta tcatacttcc atgtaagata aagcgaattg
     2461 taagattgtg gcaggaagta ggtcaagcaa tgtatgctcc tcccatcagt ggaaaaatta
     2521 tgtgtaactc aagtattaca ggaatactat tgacaagaga tggtggtaat acgagtaatg
     2581 aaaccttcag acctggagga ggaaatataa aggacaattg gagaagtgaa ttatacaaat
     2641 ataaagtagt acaaattgaa ccactaggag tagcacccac cagggcaaaa agaagagtgg
     2701 tggagagaga aaaaagagca gtgggaatag gagctatgat ctttgggttc ttaggagcag
     2761 caggaagcac tatgggcgcg gcgtcaataa cgctgacggt acaggccaga caattattgt
     2821 ctggtatagt gcaacagcaa agcaatttgc tgagagctat agaggcgcag cagcatctgt
     2881 tgcaactcac agtctggggc atcaagcagc tccaggcaag agtcctggct gtagaaagat
     2941 acctaaaaga tcaaaagttc ctaggacttt ggggctgctc tgggaaaatc atctgcacca
     3001 ctaatgtgcc ctggaactcc acttggagta ataaatcgta taaagagata tgggacaaca
     3061 tgacatggat agaatgggag agagaaatta gcaattacac aaatcaaata tatgggctac
     3121 ttacagaatc gcagaaccag caggaaagga atgaaaagga gctgttagaa ttggataaat
     3181 gggcaagtct gtggaattgg tttgacataa caaaatggct gtggtatata aaaatgttta
     3241 taataatagt aggaagttta ataggtttaa gaataatttt tgctgtgctt tctatagtaa
     3301 atagagttag gcagggatac tcacctttgt ctttccagac ccctttccat catcagaggg
     3361 aactcgacag gcccgaagga atcgaagaag gaggtggcga gcaaggcaga gacagatccg
     3421 tgcgattagt gagcggtttc ttaccaatta tctgggacga tctacggagc ctgtgcctct
     3481 tcctctacca ccgcttgaga gacttcatct cgattgtaac gaggactgtg gaacttctgg
     3541 gacgcagcag tctcaaggga ctgagacggg ggtgggaagg cctcaaatat ctggggaatc
     3601 ttctattata ttggggtcag gaactaaaaa ctagtgctat ctctctgttt gatgctgcag
     3661 cagtagcagt agcggggtgg acagataggg ttatagaagt agcacaaaga gcttggagag
     3721 ccattctcca catacctaga agaatcagac agggctttga aagggctttg ctataacatg
     3781 ggaaacaagt ggtcaaaaac atggcctcag gtcagggaaa gattagagaa aactcctcca
     3841 gcagcagagg gagtaggagc agtatctcaa gacggatcca gctaagggtc aagacaattc
     3901 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa
     3961 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
     4021 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4081 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4141 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4201 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4261 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4321 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4381 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4441 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4501 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4561 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4621 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4681 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4741 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4861 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4921 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4981 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5041 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5101 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5161 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5221 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5281 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5341 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5401 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5461 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5521 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5581 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5641 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5701 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5761 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5821 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5881 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5941 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     6001 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6061 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6121 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6181 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6241 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6301 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6361 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6421 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6481 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6541 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6601 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6661 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6721 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6781 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6841 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6901 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6961 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7021 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7081 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7141 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7201 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7261 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7321 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7381 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7441 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7501 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7561 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7621 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7681 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7741 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7801 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7861 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7921 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7981 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8041 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8101 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8161 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8221 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8281 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8341 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8401 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
