LOCUS       sequence-297219-        8471 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 8471)
  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 8471)
  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: pHM019Env
FEATURES             Location/Qualifiers
     source          1..8471
                     /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             3969..4010
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4020..4037
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4060..4077)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4066..4290
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4336..4764
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4423..4442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4633..4654
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4773..4793)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4778..5107
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4958..5093
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5020..5039
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5174..5968
                     /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(5228..5247)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5838..5857
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6144..6265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6181..6200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6235..6254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6314..6330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6314..6330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6327..6349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6338..6354
                     /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(6362..6368,6369..6386,6387..6392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6407..6428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6545..6562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6716..7304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6796..6815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7475..8266,8267..8335))
                     /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     8098..8117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8336..8440)
                     /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 gaactcctcg cagcagtaag aatcatcaag
     1021 atcttatacc agagcagtaa gtatatgtaa tgacaccttt agaaattagt gcagtagcag
     1081 gaattatagt agcattaatc ttagcaatag tagtgtggac tatagtagtt atagaaatta
     1141 agaaagtact aagagagaaa aaaatagaaa gattacttaa gagaataaga gagagagcag
     1201 aagatagtgg aaatgagagt gagggagata cggatgaatt ggcaaaactt gtggaattgg
     1261 gggactttga tccttgggtt ggggataatt tgtagtgcct cagacaactt gtgggttaca
     1321 gtttattatg gggttcctgt gtggaaagat gcagacacca ccctattttg tgcatcagat
     1381 gcccaagcac ataagacaga agtgcacaat gtctgggcca cacatgcctg tgtacccaca
     1441 gaccccaacc cacaagaaat gctcctggaa aatgtaacag aaaattttaa catgtggaaa
     1501 aataacatgg tagagcagat gcatgaagat gtaatcagtt tatgggatca aagtctaaag
     1561 ccatgtgtaa agttaactcc tctctgtgtt actttagatt gtaccagtgc taatctgagt
     1621 aacagttcca ctagcagagt agcaagcata ataggaaatg taacagagga agtaagaaac
     1681 tgctctttta atatgaccac agaactaaat gataagaggc agaagatcca tgcacttttt
     1741 tataagcttg atatagtaca aattgatgat aaaaataata atagtgagca ttataggcta
     1801 ataaactgta atacttcagt cattaagcag gcttgtccaa aggtatcctt tgatccaatt
     1861 cctatacatt attgtactcc agctggttat gcgcttttaa aatgtaatga taagaagttc
     1921 aatgggacag ggccatgtag aaatgtcagc tcagtacaat gcacacatgg aattaagcca
     1981 gtggtatcaa ctcagttgct gttaaatggt agtctagcag aagatgagat agtaatcaga
     2041 agtgaaaatc tcacagacaa tgccaagacc ataatagtgc atcttaataa atctgtagac
     2101 atcaattgta ccagaccctc caacaataca agaacaagtg tgtctatagg accaggacga
     2161 gtattctata gaacaggaga tataatagga gatataacaa aagcatattg tgagattaat
     2221 ggaacaaaat ggaatgaaac tttaacacag gtagaaggaa aattaaagga gcactttaat
     2281 aggacaataa tctttcaacc acactcagga ggagatctag agcttacaat gcatcatttt
     2341 aattgtagag gggaattttt ctattgcaat acaacaaaac tgtttaatag tccttgcctg
     2401 aaaaatgaag aaaatggaac atgtaatagc actatcatac ttccatgtaa gataaagcga
     2461 attgtaagat tgtggcagga agtaggtcaa gcaatgtatg ctcctcccat cagcggaatc
     2521 attaggtgta actcaactat tacaggaata ctattgacaa gagatggtgg taaccttaat
     2581 aacagtacta atgaaaccct cagacctgga ggaggaaata taaaggacaa ttggagaagt
     2641 gaattatata aatataaagt agtacaaatt gaaccactag gaatagcacc caccaaggca
     2701 aaaagaagag tggtggagag agaaaaaaga gcagtgggaa taggagctat gatctttggg
     2761 ttcttaggag cagcaggaag cactatgggc gcggcgtcaa taacgctgac ggtacaggcc
     2821 agacaattat tgtctggtat agtgcaacag caaaacaatt tgctgagggc tatagaggcg
     2881 cagcagcatc tgttgcaact cacagtctgg ggcattaaac agctccaggc aagagtcctg
     2941 gctgtggaaa gatacctaaa agatcaaaag ttcctaggac tttggggctg ctctgggaaa
     3001 accatctgca ccactgctgt gccctggaac gacacttgga gtaataaatc tttagaagag
     3061 atttgggaca acatgacatg gatagaatgg gagagagaaa ttagcaatta cacaaaccaa
     3121 atatatgaga tacttacaga atcgcagaac cagcaggaaa ggaatgaaaa ggatctgtta
     3181 gaattggata aatgggcaag tctgtggaat tggtttagca taacaaattg gctgtggtat
     3241 ataaggatgt ttataataat agtaggaagt ttaataggtt taagaataat ttttgctgtg
     3301 ctttctatag taaatagagt taggcaggga tactcacctt tgtctttcca gatcccttcc
     3361 catcatcaga gggaacccga caggcccgaa gaaatcgaag aaggaggtgg cgagcaaggc
     3421 agagccagat ccgtgcgctt agtgagcgga ttcttaccgc ttatctggga cgatctacgg
     3481 agcctgtgcc tcttcagcta ccaccgcttg agagacttca tcttgatcac aacgaggact
     3541 gtggaacttc tgggacacag cagtctcaag ggactgagac gggggtggga aggcctcaaa
     3601 tatctgggga atcttctatt atattggggt caggaactaa aaactagtgc catttctttg
     3661 tttgatgctg cagcaatagt aatagcggga gggacagata gggttataga agtagcacaa
     3721 agagcttgga gagccattct ccacatacct agaagaatca gacagggctt tgaaagggct
     3781 ttgctataac atgggaaaca agtggtcaaa aacatggcct cgggtcaggg acagattaga
     3841 gaagactcct ccagcagcag atggagtagg agcagtatct caagacggat ccagctaagg
     3901 gtcaagacaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgc
     3961 ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc
     4021 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     4081 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4141 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4201 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     4261 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     4321 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4381 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4441 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4501 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4561 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4621 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4681 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4741 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     4801 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4861 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4921 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4981 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5041 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5101 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     5161 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5221 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5281 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5341 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5401 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5461 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5521 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5581 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5641 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5701 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5761 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5821 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5881 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5941 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
     6001 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6061 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6121 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     6181 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6241 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6301 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6361 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6421 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6481 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6541 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6601 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6661 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6721 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6781 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6841 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6901 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6961 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7021 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     7081 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     7141 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     7201 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     7261 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7321 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7381 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7441 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     8101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8341 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8461 cacctgacgt c
//
