LOCUS       sequence-273351-        8493 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 8493)
  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 8493)
  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: pHM130Env
FEATURES             Location/Qualifiers
     source          1..8493
                     /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             3991..4032
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4042..4059
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4082..4099)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4088..4312
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4358..4786
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4445..4464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4655..4676
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4795..4815)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4800..5129
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4980..5115
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5042..5061
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5196..5990
                     /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(5250..5269)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5860..5879
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6166..6287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6203..6222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6257..6276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6336..6352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6336..6352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6349..6371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6360..6376
                     /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(6384..6390,6391..6408,6409..6414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6429..6450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6567..6584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6738..7326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6818..6837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7497..8288,8289..8357))
                     /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     8120..8139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8358..8462)
                     /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 gaacttctca gagcagtaag gatcatcaaa
     1021 atcctgtacc agagcagtaa gtaataagta tatgtaatga cacctttgga atttggtgca
     1081 atagtaggac tgatagtagc gctaatctta gcaatagtag tgtggactat agtagctata
     1141 gaagttaaga aagaactaaa acaaaggaaa gtagacaggt tagttaagag aataagagaa
     1201 agagcagaag acagtggaaa tgagagcgaa ggagacatgg atgaattggc caaacttgtg
     1261 gaaatggggg actttgatcc ttgggttggt gataatgtgt agtgcctcaa ataacttgtg
     1321 ggttacagtt tattatgggg ttcctgtgtg gagagatgca gaaaccaccc tattttgtgc
     1381 atcagatgct aaagcacagg aaagggaagt gcacaatgtc tgggccacac atgcctgtgt
     1441 gcccacagac cccaacccac aagaaatact cctagaaaat gtaacagaaa attttaacat
     1501 gtggaaaaat aacatggtag agcagatgca ggaggatgta atcagtttat gggatcaaag
     1561 tctaaagcca tgtgtaaaat taactcctct ctgcgttact ttacattgta ccaatgctaa
     1621 tttgaccaag aatgtgacca atggcaataa cacagacaat gtccttaaca tagtaggaaa
     1681 tatgacagat gaagtaagaa actgctcttt taatataacc acagaactaa gagataaaaa
     1741 gaagcaggtc catgcacttt tttataggct tgatatagta caaattgaaa atgatagtaa
     1801 taatagtagt tacaggttaa taaattgtaa tacttcagtc attaagcagg cttgtccaaa
     1861 ggtatccttt gatccaattc ctatacatta ttgtacacca gctggttatg cgattttaaa
     1921 gtgtaatgag aagaatttca atggaacagg gccatgtaaa aatgtcagct cagtacaatg
     1981 cacacatgga attaagccag tagtatcaac tcaattgctg ttaaatggta gtctagcaga
     2041 agatgagata ataatcagat ctgaaaatct cacagacaat gccaaaaaca taatagtgca
     2101 ccttaataaa tctgtagaaa tcaattgtac cagaccctct aacaatacaa gaacaagtgt
     2161 atctatagga ccaggacaag tattctatag aacaggagac ataataggag atataagacg
     2221 agcatattgt gagattaatg aaacacaatg gtataaagtt ttaagacagg taactggaaa
     2281 attaaaagag cactttaata agacaataac cttccaacca ccctcaggag gagatctaga
     2341 aactacaatg catcatttta attgtagagg ggaatttttc tattgtaata catcaaaact
     2401 atttaataat acttgcctaa atggcaccaa ggagtgtaat ggcactatca tacttccatg
     2461 caagataaag caaattataa acatgtggca gggagcagga caagcaatgt atgctcctcc
     2521 catcagtgga accattaatt gtgtatcaaa tattacagga atactactga caagagatgg
     2581 gggtagggat actaacgaga ccgagacctt cagacctgga ggaggaaata taaaggacaa
     2641 ttggagaagt gaattatata aatataaagt agtagaaatt gaaccactag gaatagcacc
     2701 caccggggca aggagaagag tggtggagag agaaaaaaga gcagtaggaa taggagctat
     2761 gatctttggg ttcttaggag cagcaggaag cactatgggc gcggcgtcca taacgctgac
     2821 ggtacaggcc agacaattat tgtctggtat agtgcaacag caaagcaatt tgctgagggc
     2881 tatagaggcg caacagcatc tgttgcaact cacagtctgg ggcattaaac agctccaggc
     2941 aagagtcctg gctgtggaaa gatacctaaa ggatcaaaag ttcctaggac tttggggctg
     3001 ctctggaaaa atcatctgca ccactaatgt gccctggaac tccacttgga gtaatagatc
     3061 tcttgaacag atttggggaa acatgacatg gacagaatgg gagagagaaa ttagcaatta
     3121 cacaaatcaa atatatgagt tacttacaga atcgcagaac cagcaggaca agaatgaaaa
     3181 ggatttgtta gaattggaca aatgggcaag cctgtggaat tggtttgaca taacaaagtg
     3241 gctgtggtat ataagaatgt ttataatgat agtaggaggg ttaataggtt taagaataat
     3301 ttttgctata ctttctatag taaaaagagt taggcaggga tactcacctt tgtcttttca
     3361 gacccctatc catcagcaga gggaacccga caggcccgaa ggaatcgaag aagaaggtgg
     3421 cgagcaaggc agagacagat ccgtgcgctt agtgagcgga ttcttagcac ttgcgtggga
     3481 cgatctacgg agcctgtgcc tcttcagcta ccaccgcttg agagacttaa tctcgattgt
     3541 agcgaggact gtggaacttc tgggacacag cagtctcaag ggactgagac aggggtggaa
     3601 aggcctcaaa tatctgggga accttctaat atattggggc caggaactaa aaactagtgc
     3661 tatttccttg cttaatgcta cagcaatagc agtaggaggg tggacagata gggttataga
     3721 gatagcacaa agagcttgga gagcctttct ccacatacct agaagaatca gacagggttt
     3781 agaaagggct ttactttaac atgggaggca agtggtcaaa aggtaaaata gtaggatggc
     3841 cgcaggtcag ggaaagaata agacgaactg agccagcagc agatggagta ggagcagtat
     3901 ctcaagacgg atccagctaa gggtcaagac aattctgcag atatccagca cagtggcggc
     3961 cgctcgagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt
     4021 ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca
     4081 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4141 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4201 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4261 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4321 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4381 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4441 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4501 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4561 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4621 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4681 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4741 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4801 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4861 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4921 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4981 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5041 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5101 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5161 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5221 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5281 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5341 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5401 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5461 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5521 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5581 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5641 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5701 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5761 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5821 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5881 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5941 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6001 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6061 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6121 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6181 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6241 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6301 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6361 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6421 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6481 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6541 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6601 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6661 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6721 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6781 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6841 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6901 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6961 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7021 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7081 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7141 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7201 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7261 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7321 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7381 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7441 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7501 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7561 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7621 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7681 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7741 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7801 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7861 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7921 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7981 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8041 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8101 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8161 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8221 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8281 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8341 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8401 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8461 cgcacatttc cccgaaaagt gccacctgac gtc
//
