LOCUS       sequence-273392-        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: pHM132F
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             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 gagcagtaag gatcatcaaa
     1021 atcctaaacc agagcagtaa gtaaatgcaa tgacaccttt ggaaattagt gcaatagcag
     1081 gattgatagt agcgctaatc gcagcaatag tagtgtggac tatagtagct atagaagtta
     1141 agaaattata taggcaaagc aaaatagaca ggttagttaa gagaataaga gaaagagcag
     1201 aagacagtgg aaatgagagt gagggagaca cagatgaatt ggccaaactt gtggaaatgg
     1261 gggactttga tccttgggtt ggtgataatt tgtagtgcct cagacaagtt gtgggttaca
     1321 gtttattatg gagttcctgt gtggaaagat gcagatacca ccctattttg tgcatcagat
     1381 gccaaagcac ctgagacaga agtgcacaat gtctgggcca cacatgcctg tgtacccaca
     1441 gaccccaacc cacaagaaat atacttggaa aatgtaacag aaaattttaa catgtggaaa
     1501 aataacatgg tagagcagat gcaggaggat gtaatcagtt tatgggatca aagtctaaag
     1561 ccatgtgtaa agttaactcc tctctgtgtt actttaaatt gtagcattgc tacttttccc
     1621 aacggcacta tgctcaagga caatagcagt atcgatgacg ctaccatagg aaatctaaca
     1681 gaggagttaa gaaactgttc ttttaatata accacagaac taatagataa gaagcaaaag
     1741 gtccatgcac ttttttataa gcttgatata gtaccaattg aagatagtag taatagtagt
     1801 acctataggt taataaattg taatacttca gtcattaagc aggcttgtcc aaaggtatcc
     1861 tttgatccaa ttcccataca ttattgtact ccagctggtt atgcgatttt caagtgtaat
     1921 gataagaatt tcaatgggac agggccatgt aaaaatgtca gctcagtaca atgcacacat
     1981 ggaattaagc cggtggtatc aactcaattg ctgttaaatg gcagtctagc agaagaagag
     2041 ataataatca gatctgaaaa cctcacaaat aatgccaaaa acataatagt gcacctcaat
     2101 gaatctgtac aaatcacttg tatcagaccc tccaacaata caagaacaag tttaactata
     2161 ggaccaggac aagtgttcta tagaacagga gacataatag gagatataag aagggcatat
     2221 tgtgagatta atggaacaaa atggaatgaa actttaagac tggtaactaa aaaattaaga
     2281 gaaagcttta ataagacaat aatctttcaa ccacccgtag gaggagatcc agaaattaca
     2341 atgcttcatt ttaattgtag aggggaattt ttctattgca atacaacaaa actgtttaat
     2401 agtacttgga gaggaaatga aaccaaaacc acgggggggt ctaataacac tcgcactgtc
     2461 attccatgca ggataaagca aattataaac atgtggcagg gagcaggaca agcaatgtat
     2521 gctcctccca tcaggggaaa cattagttgt gtatcaaata ttacaggaat actattgaca
     2581 agagatggtg gtgataataa tagtactatc gagaccttca gacctggagg aggaaatata
     2641 aaggacaatt ggagaagtga attatataaa tataaagtag tacaaattga accactagga
     2701 atagcaccct ccaaggcaag gagaagagtg gtggagagag aaaaaagagc agttggaata
     2761 ggagcgatga tctttgggtt cttaggagca gcaggaagca ctatgggcgc ggcgtcgcta
     2821 acgctgacgg tacaggccag acaattattg tctggtatag tgcaacagca aagcaatttg
     2881 ctgagggcta tagaggcgca gcagcatatg ttgcaactca cagtctgggg cattaaacag
     2941 ctccaggcaa gagtcctggc tgtggaaaga tacctaaagg atcaaaagtt cctaggactt
     3001 tggggctgct ctggaaaaat catctgcacc actaatgtgc cttggaactc cacttggagt
     3061 aataaaactt atgcagagat ttggaacaat atgacatgga cagaatggga gagagaaatt
     3121 agcaattaca caaaccaaat atatgacttg cttacagaat cgcagaacca gcaggacaaa
     3181 aatgaaaagg atttgttgga attggataaa tgggcaagct tgtggacttg gtttgacata
     3241 acacattggc tgtggtatat aaaaatattt ataataataa taggaggctt aataggttta
     3301 agaataattt ttgctgtgct ttctatagta aatagagtta ggcaggggta ctcacctttg
     3361 tctttccaga cccctaccca tcatcagagg gaacaacccg acagactcga aaaaatcgaa
     3421 gaagaaggtg gcgagcaagg cagagacaga tccgtgcgct tagtgaccgg attcttggct
     3481 ctcttctggg acgatctacg gagcctgtgc ctcttcagct accgccgctt gagagacttc
     3541 agcttgattg taacgaggac tgtggaactt ctgggacaca gcagtctcaa gggactgaga
     3601 cgggggtggg aaggcctcaa atatctgggg aatcttctgt tgtattgggg tcaggaacta
     3661 aagattagtg ctgtttcttt gtttgatgct atagccataa gagtagcggg gtggacagat
     3721 aggattatag aagtagcaca aagagcttgg agagctattc tccacatacc tagaagaatc
     3781 aggcagggct ttgaaagggc tttgttataa catgggaaac aagtggtcaa agagtagcat
     3841 agtgggatgg cctcagatca gggaaagaat aaggagaact cctccagcag cagaaggcgt
     3901 aggagcagta tctcaagacg gatcaagggt 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
//
