LOCUS       sequence-273388-        8486 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 8486)
  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 8486)
  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: pHM044Env
FEATURES             Location/Qualifiers
     source          1..8486
                     /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             3984..4025
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4035..4052
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4075..4092)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4081..4305
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4351..4779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4438..4457)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4648..4669
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4788..4808)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4793..5122
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4973..5108
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5035..5054
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5189..5983
                     /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(5243..5262)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5853..5872
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6159..6280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6196..6215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6250..6269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6329..6345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6329..6345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6342..6364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6353..6369
                     /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(6377..6383,6384..6401,6402..6407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6422..6443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6560..6577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6731..7319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6811..6830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7490..8281,8282..8350))
                     /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     8113..8132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8351..8455)
                     /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 gatcgtcaaa
     1021 atcctacacc agagcagtaa gtaaatgcaa tgacaccttt ggaaattagt gcaatagcag
     1081 gattgatagt agcgctaatc ttagcaatag tagtgtggac tatagtagct atagaagtta
     1141 agaaattata taggcaaggg aaaatagaca ggttagttaa gagaataaga gaaagagcag
     1201 aagacagtgg aaatgagagt gagggagaca cagatgaatt ggccaaactt gtggaaatgg
     1261 ggaactttga tccttgggtt ggtgataatt tgtagtgcct cagacaagtt gtgggttaca
     1321 gtctattatg gagttcctgt gtggagagat gcagatacca ccctattttg tgcatcagat
     1381 gcccaagcac atgagacaga agtgcacaat gtctgggcca cacatgcctg tgtacccaca
     1441 gaccccaacc cacaagagat acacctggaa aatgtaacag aaaattttaa catgtggaaa
     1501 aataacatgg tagagcagat gcaggaggat gtaatcagtt tatgggatca aagtctaaag
     1561 ccatgtgtaa agttaactcc tctctgtgtt actttaaatt gtagcagtgc tacttttccc
     1621 aatggcacta ccgatgacga tagcaaaatc aatggctcta ccataggaaa tctaacagag
     1681 gagttaagaa actgttcttt taatataacc acagaactaa tagataaaaa gcaaaaggtc
     1741 catgcacttt tttataagct ggatatagta ccaattgaaa gtagtaataa tagtggtcag
     1801 tataggttaa taaattgtaa tacttcaatc attaagcagg cttgtccaaa ggtatccttt
     1861 gatccaattc ctatacatta ttgtactcca gctggttatg cgatttttaa gtgtaatgat
     1921 aagaatttca atgggacagg gccatgtaaa aatgtcagct cagtacaatg cacacatgga
     1981 attaagccag tggtatcaac tcaattgctg ttaaatggca gtctggcaga agaagagata
     2041 ataatcagat ctgaaaacct cgtaaataat gccaaaacca taatagtgca cctcaatgaa
     2101 tctgtaaaaa tcacttgtac cagaccctcc aacaatacaa gaacaagtat atctatagga
     2161 ccaggacaag tgttctatag aacaggagac ataataggaa atataagaat ggcatattgt
     2221 gtgattaatg gaacaaaatg gaatgaaact ttaaaacagg taactaaaaa attaagagaa
     2281 cgctttaata agacaataat ctttcaacca cctgcaggag gagatccaga aattacaatg
     2341 tttcatttta attgtagagg ggaatttttc tattgcaata caacaaaact gtttaatagt
     2401 acttggatag gaaatgaaac cgtggagggg tctaataaca ctgtcattcc atgcaggata
     2461 aagcaaatta taaacatgtg gcagggagca ggacaagcaa tgtatgctcc tcccatcagg
     2521 ggaaacatta cttgtgtatc aaatattaca ggaatactat tgacaagaga tggtggtgat
     2581 aataatagta ctaacgagac cttcagacct ggaggaggaa atataaagga caattggaga
     2641 agtgaattat ataaatataa agtagtacaa attgaaccac taggagtagc acccaccaag
     2701 gcaaggagaa gagtggtgga gagagaaaaa agagcagtgg gaataggagc gatgatcttt
     2761 gggttcttgg gagcagcagg aagcactatg ggcgcggcgt cactaacgct gacggtacag
     2821 gccagacaat tattgtctgg tatagtgcaa cagcaaagca atttgctgag ggctatagag
     2881 gcgcagcagc atctgttgca actcacagtc tggggcatta aacagctcca ggcaagagtc
     2941 ctggctgtgg agagatacct aaacgatcaa aagttcctag gactttgggg ctgctctgga
     3001 aaaatcatct gcaccactaa tgtgccttgg aactccactt ggagtaataa atcttttaca
     3061 gagatttggg gcaacatgac atggacagaa tgggagagag aaattagcaa ttacacaaac
     3121 caaatatata gtttacttac agaatcgcag aaccagcagg acaagaatga aaaggatttg
     3181 ttggaattgg ataagtgggc aagtttgtgg acttggtttg acataacaca ttggttgtgg
     3241 tatataaaaa tatttataat gataatagga ggcttaatag gtttaagaat aatttttgct
     3301 gtgctttcta tagtaaatag agttaggcag gggtactcac ctttgtcttt ccagacccct
     3361 acccatcatc agagggaacc cgacaggccc gaaagaatcg aagaagaagg tggcgagcaa
     3421 ggcagagaca gatccgtgag attagtgagc ggattcttgg ctcttatctg ggacgatcta
     3481 cggagcctgt gcctcttcag ctaccgccgc ttgagagact tcagcttgat tgtaacgagg
     3541 actgtggaac ttctgggaca cagcagtctc aagggactga gacgggggtg ggaaggcctc
     3601 aaatatctgg ggaatcttct gttatattgg ggtcaggaac taaaaattag tgctgtttct
     3661 ttgcttgatg ctatagccat aagagtagcg gggtggacag ataggattat agaagtagca
     3721 caaagagctt ggagagcttt tatccacata cctagaagaa tcaggcaggg cttagaaagg
     3781 gctttggtat aacatgggaa acaagtggtc aaaaagtagc atagtgggat ggcctcagat
     3841 cagggaaaga ataaggagaa ctcctccagc agcagaaggc gtaggagcag tatctcaaga
     3901 cggatccagc taagggtcaa gacaattctg cagatatcca gcacagtggc ggccgctcga
     3961 gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     4021 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     4081 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4141 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4201 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4261 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     4321 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     4381 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4441 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4501 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4561 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4621 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4681 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4741 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     4801 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4861 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4921 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4981 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5041 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     5101 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     5161 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     5221 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5281 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5341 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     5401 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5461 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5521 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5581 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5641 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5701 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5761 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5821 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5881 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5941 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     6001 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6061 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6121 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     6181 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6241 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     6301 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6361 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6421 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6481 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6541 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6601 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6661 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6901 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6961 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7021 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7081 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7141 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7201 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7261 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7321 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7381 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7441 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7501 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7561 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7621 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7681 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7741 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7801 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7861 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7921 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7981 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8041 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8101 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8161 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8221 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8281 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8341 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8401 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8461 ttccccgaaa agtgccacct gacgtc
//
