LOCUS       sequence-273385-        8520 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 8520)
  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 8520)
  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: pHM153IEnv
FEATURES             Location/Qualifiers
     source          1..8520
                     /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             4018..4059
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4069..4086
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4109..4126)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4115..4339
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4385..4813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4472..4491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4682..4703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4822..4842)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4827..5156
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5007..5142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5069..5088
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5223..6017
                     /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(5277..5296)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5887..5906
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6193..6314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6230..6249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6284..6303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6363..6379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6363..6379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6376..6398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6387..6403
                     /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(6411..6417,6418..6435,6436..6441))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6456..6477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6594..6611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6765..7353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6845..6864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7524..8315,8316..8384))
                     /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     8147..8166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8385..8489)
                     /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 atcagatctg aaaatctcac
     2101 aaacaatgcc aaaaccataa tagtgcacct taatgaaact gtagaaatca attgtactag
     2161 accctccaac aatacgagaa caagtatgtc tataggacca ggacaagtat tctatagaac
     2221 aggagatata ataggagata taagacaagc atattgtgtg attaatgcaa caaaatggaa
     2281 tgaaacttta agacaggtag ttggaaaatt aagagaacat tttcataata agacaataat
     2341 ctttcaacca ccctcaggag gagatctaga aattacaaca catcatttta attgtagagg
     2401 ggaatttttc tattgcaata caacaaaact gtttaatagt acttggtaca gaaatggaac
     2461 tgtggatggg ggcaatagca caattcccaa tatcacaatt ccctgcagga taaagcaaat
     2521 tataaacatg tggcagggag taggacaagc aatgtatgct cctcccatca gtggagtaat
     2581 taattgtaca tcaaatatta caggaatact attgacaaga gatggtggta ataacaacac
     2641 tagcgaaagt gaaaccttta gacctggagg aggaaatata aaggacaatt ggagaagtga
     2701 attatataaa tataaagtag tacaaattga accattagga atagcaccca ccaaggcaaa
     2761 gagaagagtg gtggagagag aaaaaagagc agtgggaata ggagctatga tctttgggtt
     2821 cttaggagca gcaggaagca ctatgggcgc ggcgtcaata acgctgacgg tacaggccag
     2881 acaattattg tctggtatag tgcaacagca aagcaatttg ctgagggcta tagaggcgca
     2941 gcagcatttg ctgcaactca cagtctgggg cattaaacag ctccaggcaa gagtcctggc
     3001 tgtggaaaga tacctacagg atcaaaagct cctaggactt tggggctgct ctggaaaaac
     3061 catctgcacc actgctgtgc cttggaactc cacttggagt aataaatctt ttgaccaaat
     3121 ttggcaaaac atgacatgga tagaatggga gagagaaatt agcaattaca cagaccaaat
     3181 atatggtata cttacagaat cgcagaacca gcaggaaagg aatgaaaagg atttgttaga
     3241 attggataaa tgggcaagtc tgtggaattg gtttagtata acaaactggc tgtggtatat
     3301 aagaatattt ataatgatag taggaggctt aataggttta agaataattt ttgctgtgct
     3361 ttctatagta aatagagtta ggcagggata ctcacctttg tctttccaga tccccctcca
     3421 tcagcagagg gaaccagaca gactcgaaag aatcgaagaa gaaggtggcg agcaaggcag
     3481 agacagatca gtgcgcttag tgaccggatt cttagcactt gcctgggacg atctccggag
     3541 cctgtgcctt ttcagctacc accgcttgag agacttcgta ttgattgtag cgaggacagt
     3601 ggaccaggga ctgagacggg ggtgggaagg cctcaaatat ctggggaacc tcctgatata
     3661 ttggagccag gaactgaaaa ttagtgctat ctctttgttt gatactatag caatagcagt
     3721 agctgggtgg acagataggg ttatagaagt agcacaagga gcttggcgag ccattctaca
     3781 catacctaga agaatcagac agggattaga aagggctctg ctataacatg ggaaacaagt
     3841 ggtcaaaaag tagcatagtg ggatggcctc agatcaggga aagaataagg cggactcctc
     3901 cagcagcaga aggagtagga gcagtatctc aagacggatc cagctaaggg tcaagacaat
     3961 tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgcg gttcgaaggt
     4021 aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat
     4081 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     4141 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4201 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4261 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4321 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4381 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4741 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4801 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4921 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4981 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5041 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5101 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5161 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5221 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5281 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5341 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5401 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5461 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5521 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5581 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5641 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5701 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5761 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5821 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5881 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5941 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6001 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     6061 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6121 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6181 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6241 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6301 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6361 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6421 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6481 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6541 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6601 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6661 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6721 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6781 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6841 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6901 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6961 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7021 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7081 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7141 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7201 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7261 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7321 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7381 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7441 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7501 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7561 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7621 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7681 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7741 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7801 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7861 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7921 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7981 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8041 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8101 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8161 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8221 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8281 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8341 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8401 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8461 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
