LOCUS       sequence-297218-        8477 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 8477)
  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 8477)
  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: pHM073Env
FEATURES             Location/Qualifiers
     source          1..8477
                     /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             3975..4016
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4026..4043
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4066..4083)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4072..4296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4342..4770
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4429..4448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4639..4660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4779..4799)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4784..5113
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4964..5099
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5026..5045
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5180..5974
                     /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(5234..5253)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5844..5863
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6150..6271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6187..6206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6241..6260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6320..6336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6320..6336)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6333..6355)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6344..6360
                     /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(6368..6374,6375..6392,6393..6398))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6413..6434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6551..6568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6722..7310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6802..6821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7481..8272,8273..8341))
                     /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     8104..8123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8342..8446)
                     /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 caccaccgac gaggaactcc tcagagttgt gagaatcatc
     1021 aaaatcctat acgagagcag taagtaaaag tatatgtaat gacacctgtg acacctgtgc
     1081 aaattagtgc aatagtagga ctgatagtag cgctaatctc agcaatagtg gtatggacta
     1141 tagtagttat agaaattagg aaattcttaa ggcaaagaaa aatagacagg ttagttgaga
     1201 gaataagaga aagagcagaa gacagtggaa atgagagtga aggggacaca gatgaattgg
     1261 ccaaacttgt ggagatgggg ggctttgatc cttgggttgg tgataatgtg tagtgcctca
     1321 gataacttgt gggttacagt ttattatgga gttcctgtgt ggaaagatgc agataccacc
     1381 ctattttgtg catcagatgc caaagcacag gagacagaaa tgcacaatat ctgggccaca
     1441 catgcctgtg tacccacaga ccccaaccca caagaaataa ccctggagaa tgtaacagaa
     1501 aattataaca tgtggaaaaa taacatggta gagcagatgc atgaggatgt aatcagttta
     1561 tgggaccaaa gtctaaagcc atgtgtaaag ttaactcctc tctgcgttac tttagattgt
     1621 actaataatg tgacggtgaa agatgacaat agcacaaagg ccaattacag aaactctcca
     1681 gtaggaaaca taacagatga attaagaaac tgttctttta atatgaccac agaacttagg
     1741 gataagaagc agaaggcata tgcacttttt tataagcttg acatagtatc actgaaaaag
     1801 aatagtagtg agtataggtt aataaattgt aatacttcag taattaagca ggcttgtcca
     1861 aaggtatcct ttgatccaat tcctatacat tattgtactc cagctggtta tgtgatttta
     1921 aagtgtaagg ataagaagtt caatgggaca gggccatgtg aaaatgtcag ctcagtacaa
     1981 tgcacacatg gaattaagcc agtggtatca actcaattgc tgttaaatgg tagtctagca
     2041 gaagacgaga tagtaatcag ctctgaaaat atcacaaaca atgccaaaac cataatagtg
     2101 caccttaatg aatctgtaga aatcaattgt accagaccct ccaacaatac aagaaaaagt
     2161 atacgtatag gaccaggaca agtcttctat caaacaggag acataacagg agatataaga
     2221 aaagcatatt gtgagattaa tggaacaaaa tggaataaaa ctttagaaca ggtaaagaga
     2281 aaattaggag agcactttaa gaagaacata acctttcagc caccctcagg aggagatcta
     2341 gaaattacaa tgcatcattt tatttgtaga ggggaatttt tctattgcaa tacatcacaa
     2401 ctgtttgata gcacaaaaaa tatcacagga aatatcacac ttccatgtag gataaaacaa
     2461 attataaaca tgtggcaggg aacaggacag gcaatgtatg ctcctcccat cgagggagta
     2521 attaattgtg tatcaaatat tacaggaata ctattgacaa gagatggtgg tattaataat
     2581 accaacgaga ccttcaggcc tgggggagga gacatgaggg acaattggag aagtgaacta
     2641 tataaatata aagtagtaag aattgaacca ctaggaatag cacccaccaa ggcaaagaga
     2701 agagtggtgg agagggaaaa aagagcagtg ggaataggag ctatgatctt tgggttctta
     2761 ggagcagcag gaagcactat gggcgcagcg tcaataacgc tgacggtaca gaccagacaa
     2821 ttattgtctg gtatagtgca acagcaaagc aatttgctga gggctataga ggcgcaacag
     2881 catatgttgc aactcacagt ctggggcatt aaacagctcc aggcaagagt cctggctgtg
     2941 gaaagatacc taaaagatca aaggttccta gggctttggg gctgctctgg aaaaaccatc
     3001 tgcaccacta ctgtaccctg gaacacaact tggagtaata aatcttatga cgagatctgg
     3061 ggcaacatga catggataca atgggagagg gaaattggca attacacaaa ccaaatatat
     3121 gatatactta cagaatcgca gaaccagcag gaaaagaatg aacaggattt gttggcattg
     3181 gatacatggg caagtctgtg gaactggttt gacataacaa attggctgtg gtatataaaa
     3241 atatttatca tgatagtagg aggtttaata ggtttaagaa taatttttgc tgtgctttct
     3301 atagtaaata gagttaggca gggatactca cctttatctt tccagacccc tatccatcag
     3361 cagagggaac ccgacaatcc cgaaggaatc ggagaaggag gtggagagca aggcagagac
     3421 agatccgtgc gattagtgag cggattctta gctcttgcgt gggacgatct acggagcctg
     3481 tgcctcttca gctaccaccg cttgagagac ttcatctcga ttgcagcgag gactgtggaa
     3541 cttctgggac ggagcagtct caagggactg agacgagtgt gggaagccct caaatatctg
     3601 gggaatcttc tgttatattg gggtcaggaa ctaaagatta gtgctatttc tttgtttgat
     3661 actatagcaa tagcagtagc ggggtggaca gataggatta tagaagtagc acaaagaatt
     3721 tggagagcca ttctcaacgt acctagaaga atcagacagg gtttagaaag atttttgcta
     3781 taacatggga ggcaaatggt caaaaagtag catagtggga tggcctcagg tcagggaaag
     3841 aataaggaaa actcctccag cagcagaagg agtaggagca gtatctcaag acggatccag
     3901 ctaagggtca agacaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg
     3961 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     4021 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     4081 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4141 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4201 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4261 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4321 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4381 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4441 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4501 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4561 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4621 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4681 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4741 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4801 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4861 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4921 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4981 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5041 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5101 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     5161 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     5221 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5281 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5341 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5401 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5461 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5521 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5581 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5641 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5701 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5761 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5821 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5881 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5941 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga
     6001 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6061 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6121 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6181 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6241 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6301 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6361 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6421 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6481 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6541 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6601 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6661 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6901 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7141 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8461 aagtgccacc tgacgtc
//
