LOCUS       sequence-273387-        8444 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 8444)
  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 8444)
  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: pHM024Env
FEATURES             Location/Qualifiers
     source          1..8444
                     /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    2724..2957
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3480..3506
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3942..3983
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3993..4010
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4033..4050)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4039..4263
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4309..4737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4396..4415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4606..4627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4746..4766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4751..5080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4931..5066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4993..5012
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5147..5941
                     /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(5201..5220)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5811..5830
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6117..6238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6154..6173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6208..6227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6287..6303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6287..6303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6300..6322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6311..6327
                     /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(6335..6341,6342..6359,6360..6365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6380..6401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6518..6535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6689..7277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6769..6788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7448..8239,8240..8308))
                     /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     8071..8090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8309..8413)
                     /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 taatgcaaac tttagagata tatgcaatag
     1081 tagcattagt aatagtagga ataattgcaa tagttgtgtg gactatagta cttatagaat
     1141 ataagaaaat attaagacaa agaagaatag acaggttact tgaaagaata agagaaagag
     1201 cagaagacag tggcaatgag agtgaggggg atcaggagga attacttatg gaaatggggg
     1261 aaatggggca tcttattcct ggggatattg atgatctgta gcgctgcaga caatttgtgg
     1321 gtcacagtct attatggggt acctgtgtgg agagatgcaa acaccactct attttgtgca
     1381 tcagatgcta aagcatatga aagagaagta cataatgttt gggccacaca tgcctgtgta
     1441 cccacagacc ccaacccaca agaattggta ttggaaaatg tgacagaaga gtttaatatg
     1501 tggaaaaata acatggtaga acagatgcat gaggacataa tcagtttatg ggatcaaagc
     1561 ctaaaaccat gtgtaaaatt aaccccactc tgtgtcactt taaattgcac tgacgctacc
     1621 agtggtaata ataccagtgg taatataacg ggggagggag gaatgaaaaa ctgctctttt
     1681 aatgtcacca cagacataaa agataagaga atagaaaaag cattgtttta taaagatgat
     1741 ttagtacaaa cagggaagaa taatactatt tataggttga tgagctgtaa cacctcagtc
     1801 attacccaag cctgtccaaa ggtatccttt gagccaattc ccatacatta ttgtaccccg
     1861 gctggttatg cgattcttaa gtgtaataat aagacattta atggattagg gcgatgtaca
     1921 aatgtcagca cagtactatg tacacatgga attaggccag tagtatcaac tcaactgatg
     1981 ttaaatggca gtttagcaga agaagaggta gtaattagat ctgaaaattt cacaaacaat
     2041 gctaaaacca taatagtaca gctgaaagaa gctgtaaaaa ttaattgcac aagacccaac
     2101 aacaatacaa gaaaaagtat acatatgggg caagggaaag catggtatac aacaggagag
     2161 ataataggag acataagaca agcatattgt acaattaaca aaacaaaatg gaatgacact
     2221 ttaaaacaga tagctgaaaa attaagaaga caatttggga ataaaacaat agtctttaat
     2281 cagccctcag gaggggaccc agagattgta atgcacagtt ttaattgtgg aggggaattt
     2341 ttctactgta atacatcaca actatttaat agtacttggg atggaaatag tactagggag
     2401 aatagcactg gacagttatt cacactccca tgcagaataa aacaaattat aaacatgtgg
     2461 cagaaagtag gaaaagcaat gtatgcccct cccatcgcgg gcccaattaa ctgtttatca
     2521 aacattacag ggctgttatt aacaagagat ggtggtggta ataatacaaa taatgagacc
     2581 ttcagaccag gaggaggaga tatgagggac aattggagaa gtgaactata caaatataaa
     2641 gtagtaagag ttgaaccatt aggactggca cccaccaggg caaagagaag agtggtgcag
     2701 agagaaaaaa gagcagtggg actaggagca ttgttcattg ggttcttggg agcagcagga
     2761 agcactatgg gcgcagcgtc agtgacgctg acggtacagg ccagactatt attgtctggt
     2821 atagtgcaac agcagaacaa tctgctgagg gctattgagg cgcaacagca tctgttgcaa
     2881 ctcacagtct ggggcatcaa gcagctccag gcaagagtcc tggctgtgga aagataccta
     2941 agggatcaac agctcctagg aatgtggggt tgctctggaa aactcatttg ccccacctct
     3001 gtgccttgga atgctagttg gagcaataaa tctctggaag atatttggaa taacatgacc
     3061 tggaggcaat gggagacaga aattgacaat tacacaggct taatatacac cttacttaca
     3121 gaatcgcaga accaacagga aaagaatgaa caagaattat tagaattgga taaatgggca
     3181 aatttatgga attggtttga catatcaaac tggctgtggt atataagaat attcataatg
     3241 atagtaggag gcttgataag tttaagaata gtttttactg tactttctat aataaataga
     3301 gttaggcagg gatactcacc attgtcgttg cagacccacc tcccagctca gaggggaccc
     3361 gacaggccag aaggaatcga agaagaaggt ggagagagag acagagacag atccggacga
     3421 ttagtgggtg gattcttgac acttatctgg gtcgacctga ggagcctgtt cctctacagc
     3481 taccaccgct tgagagactt actcttgatt gtaacgagga ttgtggaact tctgggacgc
     3541 agggggtggg aactcctcaa gtactggggg aatctcctac agtattgggg tcaggaacta
     3601 aagaatagtg ctattagctt gctcaacacc acagccatag cagtagctga ggggacagat
     3661 aggcttatag aattagtgca attaactttt agagctattc ttcacatacc tagaagaata
     3721 agacagggct tagaaaggac tttgctataa gatgggtggc aagtggtcaa aacgcagtag
     3781 gagtgaatgg cctgctgtaa gggaaagaat gcaacgagtt gagccagcag cagaaggggt
     3841 gggagcagta tctcgagacg gatccagcta agggtcaaga caattctgca gatatccagc
     3901 acagtggcgg ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc
     3961 ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa
     4021 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     4081 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     4141 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     4201 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     4261 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     4321 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4381 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4441 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4501 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4561 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4621 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4681 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4741 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4801 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4861 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4921 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4981 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     5041 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     5101 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     5161 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     5221 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5281 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     5341 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5401 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5461 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5521 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5581 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5641 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5701 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5761 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5821 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5881 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5941 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6001 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6061 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6121 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6181 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6241 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6301 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6361 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6421 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6481 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
