LOCUS       sequence-273384-        8447 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 8447)
  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 8447)
  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: pHM069Env
FEATURES             Location/Qualifiers
     source          1..8447
                     /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             3945..3986
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3996..4013
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4036..4053)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4042..4266
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4312..4740
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4399..4418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4609..4630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4749..4769)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4754..5083
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4934..5069
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4996..5015
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5150..5944
                     /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(5204..5223)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5814..5833
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6120..6241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6157..6176)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6211..6230
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6290..6306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6290..6306)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6303..6325)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6314..6330
                     /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(6338..6344,6345..6362,6363..6368))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6383..6404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6521..6538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6692..7280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6772..6791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7451..8242,8243..8311))
                     /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     8074..8093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8312..8416)
                     /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 tcaagcttct cctatggcag
      961 gaagaagcgg aagcaccgac gaggaactcc tagcagcagt aaggatcatc aaattcttat
     1021 accaaagcag taagtaataa gtatatgtaa tgacaccttt ggaaattagt gcaatagtag
     1081 gactaatagt agcgctaatt gcagcaatag tagtgtggat catagtagct ttagaattta
     1141 ggaaaatact aatacaaaga aaaatagaca ggttagttaa aagaataaga gaaagagaag
     1201 aagacagtgg aaatgagagt gaaggagaca cagatgaatt ggccaaactt gtggaaatgg
     1261 gggattttga tccttgggtt ggtgataatt tgtagtgcct cagacaaatt gtgggttaca
     1321 gtttactatg gggttcctgt gtggagagat gcagatacca ccctattttg tgcatcagat
     1381 gctaaagcac atgatacaga aatgcacaat gtctgggcca cacatgcctg tgtacccaca
     1441 gaccccaacc cacaagaaat acccctggaa aatgtaacag aaaattttaa catgtggaaa
     1501 aataacatgg tagagcagat gcatgaggat gtaatcagtt tatgggacca aagtctaaag
     1561 ccatgtgtaa agttaactcc tctttgcgtt acgttaaatt gtacctgggc taattggacc
     1621 aatgccaatg acagtgacac agtaacagat gaagtaagaa actgcacttt taatatgacc
     1681 acagtaatac gagataagaa gcagcaggtt catgcacttt tttataggct tgatatagtg
     1741 ccaattgaaa agaatagtag tacttatagg ttaataaatt gtaatacttc agtcattaag
     1801 caggcttgtc caaaaatatc ctttgatcca attcctatac attattgtac tccagctggt
     1861 tatgcgattt taaagtgtaa tgataagaat ttcaatggga cagggccatg taaaaatgtc
     1921 agctcagtac aatgcacaca tggaattaag ccagtggtat caactcaatt gctgttaaat
     1981 ggcagtctag cagaagaaga gataataatc agatctgaaa atctcacaga caatgctaaa
     2041 aacataatag tgcaacttaa tgaatctgta gaaatcaatt gtaccagacc ctccaacaat
     2101 acaagaacaa gtgtagctat aggaccagga caagtatggt atagaacaga agacataata
     2161 ggaaatataa ggaaagcata ttgtgaaatc aatggaacaa actggtataa agttttaaaa
     2221 agggtagctg aaaaattcaa aaggcacttt aagaataaaa caatatcctt tcaaccaccc
     2281 tcaggaggag atctagaaat tacaatgcat cattttactt gtagagggga atttttctat
     2341 tgcaatacaa cagaactgtt taattataca gaaaatgaag ccacgaaggg ggataataca
     2401 actatcagac ttccatgtaa gataaagcaa attataaaca tgtggcaggg agtaggacaa
     2461 gcaatgtatg ctcctcccat cagtggaagc attagttgta tatcaaatat tacaggaata
     2521 ctattgacaa gagatggtgg tgataataat gttaatgaga ccttcagacc tggaggagga
     2581 aatataaagg acaattggag aagtgaatta tataaatata aagtagtaca aattgaacca
     2641 ctaggaatag cacccaccaa ggcaaagaga agagtggtgg agagagagaa aagagcagtg
     2701 ggaataggag ctatgatctt tgggttctta ggagcagccg gaagcactat gggcgcggcg
     2761 tcaataacgc tgacggtaca agccagacaa ttattgtctg ggatagtgca acagcaaagc
     2821 aatttgctga gggctataga ggcgcagcag catctgttgc aactcacagt ctggggcatc
     2881 aaacagctcc aggcaagagt cctggctgtg gaaagatacc taaaagatca aaagttccta
     2941 ggactttggg gctgctctgg aaaaatcatc tgcaccactg ctgtgccctg gaactccact
     3001 tggagtaata aatcttatga ggagatttgg gacaacatga catggataga atgggagaaa
     3061 gaaattagca attacacaaa taaaatatac gagctaatca cagaatcgca gaaccagcag
     3121 gacaaaaatg aaagggattt attggaattg gataaatggg caagtctgtg gaattggttt
     3181 gacataacaa actggctgtg gtatataaaa atatttataa tgatagtagg aggcttaata
     3241 ggtttaagaa taatttttgc tgtgctttct atagtgaata gagttaggca gggatactca
     3301 cctttgtctc tccagaccct tttccaccac cagagggaac ccgacaggcc cgaaggaatc
     3361 gaagaagaag gtggcgagcg aggcaaagac agatccgtgc gattagtgag cggattctta
     3421 tctcttgctt gggacgatct acggaacctg tgcctcttca gctaccaccg cttgagagac
     3481 ttcagcttga ttgtagtgag gactgtggaa cttctgggac gcagcagtct caagaaattg
     3541 aaacaggggt gggaaggcct caaatatctg gggaatcttc tggtctattg gattcaggaa
     3601 ctaaaaacca gtgctatttc tttgtgtgat actgcggcaa tagcagtagc ggggtggaca
     3661 gatagggtga tagaagtagc acaaagagct tggagagcct ttctccacat acctagaaga
     3721 attagacagg gcttcgagag ggctttgcta taacatggga agcaagtggt caaaaagtag
     3781 catagtggga tggcctgaga ccagggaaag aataaggcga gcacctccag cagcagcagg
     3841 agtaggagca gtatctcaag acggatccag ctaagggtca agacaattct gcagatatcc
     3901 agcacagtgg cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta
     3961 accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt
     4021 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4081 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4141 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4201 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     4261 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     4321 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4381 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4441 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4501 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4561 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4621 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4681 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4741 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4801 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4861 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4921 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4981 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     5041 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     5101 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5161 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5221 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5281 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     5341 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5401 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5461 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5521 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5581 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5641 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5701 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5761 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5821 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5881 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5941 ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6001 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6061 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     6121 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6181 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6241 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6301 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6361 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6421 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6481 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6541 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6601 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6661 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6721 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6781 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6841 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6901 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6961 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7021 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7081 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7141 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7201 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7261 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7321 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7381 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7441 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7501 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7561 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7621 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7681 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7741 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7801 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7861 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7921 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7981 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8041 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8101 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8161 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8221 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8281 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8341 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8401 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
