LOCUS       sequence-275052-        8521 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 8521)
  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 8521)
  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: pHM153BEnv
FEATURES             Location/Qualifiers
     source          1..8521
                     /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             4019..4060
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4070..4087
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4110..4127)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4116..4340
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4386..4814
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4473..4492)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4683..4704
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4823..4843)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4828..5157
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5008..5143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5070..5089
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5224..6018
                     /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(5278..5297)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5888..5907
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6194..6315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6231..6250)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6285..6304
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6364..6380)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6364..6380)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6377..6399)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6388..6404
                     /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(6412..6418,6419..6436,6437..6442))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6457..6478
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6595..6612)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6766..7354)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6846..6865)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7525..8316,8317..8385))
                     /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     8148..8167
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8386..8490)
                     /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 gatctcctca gagcagtaag gatcatcaaa
     1021 atcctatacc aaagcagtaa gtactaaata catgtaatga cacctttgga aattagtgca
     1081 atagtaggac tgatagtagc gctaatctta gcaatagtag tgtggactat agtagctata
     1141 caaatcaaac aattactaag acaaaggaaa atagacaggt taattaagag aataagagaa
     1201 agagaagaag acagtggaaa tgagagtgaa ggagacaccg atgacttggc caaacttttg
     1261 gaaatggggg actttgctcc ttgggttggt gatgatatgt agtgcctcag acaacttatg
     1321 ggttacagtt tattatgggg ttcctgtgtg gaaagatgca gataccacct tattttgtgc
     1381 atcagatgcc aaagcacagg acccagaagt gcacaatgtc tgggccacac atgcctgtgt
     1441 acccacagac cccaacccac aagaaatgca cctggtaaat gtaacagaga attttaacat
     1501 gtggaaaaat aacatggtag aacagatgca ggaggatgta atcagtttat gggaccaaag
     1561 tctacagcca tgtgtaaagt taacccctct ctgcgttact ttgaactgta cccaggctaa
     1621 tccgactaat gtcactaaga acgctgtccc aggagctgga gctaacttag aaaatgtaac
     1681 agaagaaata agaaactgtt cttttaatat gaccacagaa ctaagagata agaagaagaa
     1741 ggtctatgca ctcttttata agattgatat agtacaactt ggaagtaata ataatagtag
     1801 taataataag agtagtaatt ctagtgatta tagtgagtat aggttaataa attgtaatac
     1861 ttcagtcatt aaacaggctt gtccaaaggt atcttttgat ccaattccta tacattattg
     1921 tactccagct ggttatgcgc ttctaaagtg taatgagaag aatttcactg ggacagggcc
     1981 atgtaaaaat gtcagctcag tacaatgcac acatggaatt aagccagtgg tatcaactca
     2041 attactatta aatggcagtc tagcagaagg agacataata attagatctg aaaatctcac
     2101 aaacaatgcc aaaaccataa tagtgcacct taataaaact gtagaaatca attgtactag
     2161 accctccaaa aaaatgagaa caagtttccg tataggacca ggacaagtat tctacagaac
     2221 aggagatata ataggagata taagacaagc acattgtgtg attaatgcaa caaaatggaa
     2281 tgaaacttta agacaggtag ctagaaaatt aagagaacat tttcctaata agacaataat
     2341 ctttcgacca ccctcaggag gagatctaga aattacaaca catcatttta attgtagagg
     2401 ggaatttttc tattgcaata catcaaaact gtttagaagt acttggacag gaaatgaaac
     2461 tgtggattgg aaaggaaatg gatctgtgag tgatgacaat atcacaattc cctgcaggat
     2521 aaagcaaatt ataaacatgt ggcagggagt aggacaagca atgtatgctc ctcccatcag
     2581 tggagtaatt aattgtacat caaatattac aggaatacta ttgacaagag atggtggtac
     2641 gaatagcaac actagcgaaa cctttagacc tggaggagga aatataaaag acaattggag
     2701 aagtgaatta tataaatata aagtagtaca aattgaacca ttaggaatag cacccaccaa
     2761 ggcaaagaga agagtggtgg agagagaaaa aagagcagtg ggaataggag ctatgatctt
     2821 tgggttctta ggagcagcag gaagcactat gggcgcggcg tcaataacgc tgacggtaca
     2881 ggccagacaa ttattgtctg gtatagtgca acagcaaagc aatttgctga gggctataga
     2941 ggcgcagcag catctgctgc aactcacagt ctggggcatt aaacagctcc aggcaagagt
     3001 cctggctgtg gaaagatacc tacaggatca aaagctccta ggactttggg gctgctctgg
     3061 aaaaaccatc tgcaccactg ctgtgccttg gaactccact tggagtaata aatcttttga
     3121 ccaaatttgg caaaacatga catggataga atgggagaga gaaattagca attacacaga
     3181 ccaaatatat ggtatactta cagaatcgca gaaccagcag gaaaggaatg aaaaggattt
     3241 gttagaattg gataaatggg caagcctgtg gaattggttt agcataacaa actggctgtg
     3301 gtatataaga atatttataa tgatagtagg aggcttaata ggtttaagaa taatttttgc
     3361 tgtgctttct atagtaaata gagttaggca gggatactca cctttgtctt tccagatccc
     3421 cctccatcag cagagggaac cagacagact cgaaagaatc gaagaagaag gtggcgagca
     3481 aggcagagac agatcagtgc gcttagtgac cggattctta gcacttgcct gggacgatct
     3541 ccggagcctg tgccttttca gctaccaccg cttgagagac ttcatcttga ttgtagcgag
     3601 gacagtggac cagggactga gacgggggtg ggaaggcctc aaatatctgg ggaacctcct
     3661 gatatattgg agccaggaac tgaaaattag tgctatctct ttgtttgata ctatagcaat
     3721 agcagtagct gggtggacag atagggttat agaagtagca caaggagctt ggcgagccat
     3781 tctacacata cctagaagaa tcagacaggg attagaaagg gctctgctat aacatgggaa
     3841 acaagtggtc aaaaagtagc atagtgggat ggcctcagat cagggaaaga ataaggcgga
     3901 ctcctccagc agcagaagga gtaggagcag tatctcaaga cggatcaagg gtcaagacaa
     3961 ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgc ggttcgaagg
     4021 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca
     4081 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     4141 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4201 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4261 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4321 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     4381 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4441 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4501 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4561 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4621 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4681 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4741 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4801 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4861 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4921 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4981 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5041 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5101 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5161 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5221 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5281 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5341 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5401 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5461 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5521 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5581 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5641 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5701 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5761 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5821 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5881 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5941 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6001 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     6061 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6121 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6181 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6241 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6301 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6361 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6421 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6481 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6541 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6601 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6661 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6721 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6781 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6841 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6901 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6961 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7021 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7081 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7141 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7201 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7261 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7321 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7381 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7441 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7501 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7561 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7621 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7681 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7741 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7801 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7861 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7921 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7981 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8041 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8101 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8161 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8221 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8281 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8341 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8401 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8461 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8521 c
//
