LOCUS       sequence-275055-        8462 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 8462)
  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 8462)
  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: pHM053Env
FEATURES             Location/Qualifiers
     source          1..8462
                     /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    2742..2975
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3498..3524
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3960..4001
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4011..4028
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4051..4068)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4057..4281
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4327..4755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4414..4433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4624..4645
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4764..4784)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4769..5098
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4949..5084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5011..5030
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5165..5959
                     /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(5219..5238)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5829..5848
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6135..6256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6172..6191)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6226..6245
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6305..6321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6305..6321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6318..6340)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6329..6345
                     /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(6353..6359,6360..6377,6378..6383))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6398..6419
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6536..6553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6707..7295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6787..6806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7466..8257,8258..8326))
                     /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     8089..8108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8327..8431)
                     /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 aggcagtaag actcatcaag
     1021 cttcttcatc aaagcagtaa gtaatatatg taatgcaaac tttagagata tattcaatag
     1081 tagcattggt agtagtagga ataattgcaa tagttgtgtg gactatagta cttatagaat
     1141 ataagaaaat agtaagacaa aaaagaatag acaggttact tgaaagaata agggaaagag
     1201 cagaagacag tggcaatgag agtgaggggg atcaggagga attacttatg gaaatggggg
     1261 aaatggggca tcttattcct ggggatattg atgatctgta gcgctgcaga caagttgtgg
     1321 gtcacagtct attatggggt acctgtgtgg aaagatgcaa acgccactct attttgtgca
     1381 tcagatgcta aagcatatga aagagaagta cataatgttt gggccacaca tgcctgtgta
     1441 cccacagacc ccaacccaca agaattggta ttgggaaatg tgacagaaga gtttaatgtg
     1501 tggaaaaata acatggtaga acagatgcag gaggacataa tcagtttatg ggatcaaagc
     1561 ctaacaccat gtgtaaaatt aaccccactc tgtgtcactc taaactgcac tgaccttgcc
     1621 actaacgcta ccagtggtaa tgctaccagt ggtaatataa cgggggaggt agtaatgaaa
     1681 aactgctctt tcaatgtcac cacagacaca acagaaaaga caaaagaaaa agcattgttt
     1741 tataaagatg atttagtaca aatagggaat aatactattt ataggttgat gagatgtaac
     1801 acctcagtca ttacccaggc ctgtccaaag gtatcctttg agccaattcc catacattat
     1861 tgtaccccgg ctggttatgc gattcttaag tgtaataata aaacatttaa tggaaaaggg
     1921 ctatgtacaa atgtcagcac agtacaatgt actcatggaa ttaggccagt agtatcaact
     1981 caactgctgt taaatggcag tttagcagaa gaagaggtag taattagatc tgaaaatctc
     2041 acaaacaatg ctaaaaccat aatagtacag ctgaaagaag ctgtagaaat taattgcaca
     2101 agacccaaca acaatacaag aaaaagtata agtgtggggc cagggagagc atggtttaca
     2161 acaggagaga taataggaga cataagacaa gcatattgta aaattaacag agcaaaatgg
     2221 aataacactt taaaacagat agctaaaaaa ttaagagttc aatttgggaa taaaacaata
     2281 gtctttaatc actcctcagg aggggaccca gagattgtaa tgcacagttt taattgtgga
     2341 ggggaattct tctactgtaa tacatcacaa atatttaata gtacttggaa tggaaatagc
     2401 actattggga atagcactga agagtcaaat agcacattca cactcccatg cagaataaaa
     2461 caaattataa atatgtggca gaaaatagga aaagcaatgt atgcccctcc catcgcgggt
     2521 ccaattaact gttcatcaaa cattacaggg ctgttattaa caagagatgg tggtgataat
     2581 aatacaacta aagagatctt cagaccagga ggaggagata tgagggacaa ttggagaagt
     2641 gaactataca aatataaagt agtaagagtt gaaccattag gactggcacc caccaaggca
     2701 aagagaagag tggtgcagag agaaaaaaga gcagtgggac taggagcgtt gttcattggg
     2761 ttcttgggag cagcaggaag cactatgggc gcagcgtcag tgacgctgac ggtacaggcc
     2821 agactattat tgtctggtat agtgcaacag cagaacaatt tgctgagggc tattgaggcg
     2881 caacagcata tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagagtcctg
     2941 gctgtggaaa gatacctaaa ggatcaacag ctcctaggac tttggggttg ctctggaaaa
     3001 ctcatttgcc ccacctctgt gccttggaat gctagttgga gcaataaatc tctggaagat
     3061 atttgggaga acacgacttg gatgcaatgg gagagagaaa ttgacaatta cacaggctta
     3121 atatacaccc tacttacaga atcgcagaac caacaggaaa agaatgaaca agaattatta
     3181 gaattggata aatgggcaaa tttatggaat tggtttgaca tatcaaactg gctgtggtat
     3241 ataagaatat tcataatgat agtaggaggc ttgataggtt taagaatagt ttttactata
     3301 ctttctataa taaatagagt taggcaggga tactcaccat tgtcgttgca gacccacctc
     3361 ccagctcaga ggggacccga caggccagaa ggaatcgaag aagaaggtgg agagagagac
     3421 agaaacggat ccggacgatt agtgggtgga ttcttgacac ttatctgggt cgacctgagg
     3481 agcctgttcc tctacagcta ccaccgcttg agagacttac acttgattgt aacgaggatt
     3541 gtggaacttc tgggacgcag ggggtgggaa ctcctcaagt attggtggaa tctcctacag
     3601 tattggagtc aggaactaaa gaatagtgct actagcttgc tcaacaccac agccatagta
     3661 gtagctgagg ggacagatag gtttatagaa ttagtgcaat tagcttgtag agctattctc
     3721 cacataccta gaagaataag acagggctta gaaaggactt tgctataaga tgggtggcaa
     3781 gtggtcaaaa cgaagtaaga atgaatggcc tgctgtaagg gaaagaatgc aacgagaaga
     3841 gccagcagca gaaggggtgg gagcagtatc tcgagacgga tccagctaag ggtcaagaca
     3901 attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaag
     3961 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc
     4021 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     4081 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4141 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4201 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4261 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     4321 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4381 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4441 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4501 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4561 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4621 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4681 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4741 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4801 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4861 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4921 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4981 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     5041 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     5101 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     5161 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5221 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5281 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5341 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5401 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5461 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5521 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5581 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5641 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5701 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5761 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5821 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5881 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5941 tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc
     6001 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6061 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6121 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6181 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6241 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6301 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6361 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6421 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6481 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6541 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6601 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6661 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6721 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6781 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6841 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6901 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6961 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7021 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7081 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7141 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7201 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7261 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7321 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7381 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7441 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7501 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7561 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7621 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7681 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7741 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7801 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7861 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7921 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7981 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8041 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8101 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8161 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8221 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8281 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8341 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8401 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8461 tc
//
