LOCUS       sequence-288125-        8474 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 8474)
  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 8474)
  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: pHM141Env
FEATURES             Location/Qualifiers
     source          1..8474
                     /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"
     misc_feature    2754..2987
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3513..3539
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3972..4013
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4023..4040
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4063..4080)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4069..4293
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4339..4767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4426..4445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4636..4657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4776..4796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4781..5110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4961..5096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5023..5042
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5177..5971
                     /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(5231..5250)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5841..5860
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6147..6268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6184..6203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6238..6257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6317..6333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6317..6333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6330..6352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6341..6357
                     /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(6365..6371,6372..6389,6390..6395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6410..6431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6548..6565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6719..7307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6799..6818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7478..8269,8270..8338))
                     /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     8101..8120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8339..8443)
                     /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 gaagcggaga ccgcgacgaa gacctcatca agacagtccg actcatcaag
     1021 cttctctatc aaagcagtaa gtagtatatg tgatgatacc tttaacagta gcaatagtag
     1081 cattagtagt agcagcaata ctagcaataa ttgtatggac catagtattc ttagaatata
     1141 ggaaaatatt aagacaaaga aaaatagaca ggttaattga tagaataagt gaaagagcag
     1201 aagacagtgg caatgaaagt gaaggagacc aggaagaatt gtcagcgctt atggaaatgg
     1261 ggcatcatgc tccttgggat gttaatgata tgtagtgctg cagaacaatt gtgggttaca
     1321 gtatattatg gggtacctgt gtggaaagaa gcaatcacca ctttattttg tgcatcagat
     1381 gctaaagcat atgacacaga ggtacataat gtttgggcca cacatgcctg tgtacccaca
     1441 gaccccaacc cacaagaagt aaggatggaa aatgtgacag aagattttaa catgtgggaa
     1501 aataatatgg tagaacagat gcatgaagac ataatcagtt tatgggatca aagtttaaag
     1561 ccatgtgtaa aattaacccc actctgtgtt actttagagt gcgatgatta taccaccaat
     1621 agtactgtta ccaatagtag tagtgagaaa acaggggaaa aaggagaaat aaaaaactgc
     1681 tctttcaatg tcaccacaac aataagagat aaggtggaaa aagaacaagc actgttttat
     1741 aaacttgatg tagtaccaat agaaaatagt agtaatagta ccaaatatag gttaagaaat
     1801 tgtaatacct cagtcattac acagacttgt ccaaaggtat cctttgagcc aattcccata
     1861 cactattgtg ccccggctgg ttttgcgatt gtaaagtgta acaataagac attcagtgga
     1921 aaaggaccat gtaccaatgt cagcacagta caatgtacac atggaattag gccagtagtg
     1981 tcaactcagc tgttgttaaa tggcagtcta gcagaagaag acatagtaat tagatctaag
     2041 aatttcacag ataattcaaa aaccataata gtacaactga acaaatccat agcaattaat
     2101 tgtacaagac ccaacaacaa cacaagaaaa agtatacatt taggaccagg gcaagcatgg
     2161 tatacaacag gagccataat aggagatata agacaagcac attgtaacat tagtaaaata
     2221 gaatggaata atactttaag acaggtagtt aaaaaattaa aagaaaaatt taatgcaaca
     2281 acaatattct ttaatcaatc ctcaggggga gacccagaaa ttgtaatgca cagctttaat
     2341 tgtggagggg aatttttcta ctgcaataca acacaactat ttaatagttc ttggaataat
     2401 agtagtttga atagtactga gggtaatgga gggtcaaatg acactatcac actcccgtgt
     2461 agaataaaac aaattataaa catgtggcag gaagtaggaa aagcaatgta tgcccctccc
     2521 attcaaggac aaattagatg ctcctcaaat attacaggac tattattagt aagagatggt
     2581 ggtaataaca ggacgggcgg cagtgagacc ttcagacctg gaggaggaaa tatgaaggac
     2641 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggaatagca
     2701 cccaccaggg caaggagaag agtggtgcag agagaaaaaa gagcagtggg actaggagct
     2761 ttgttccttg ggttcctggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     2821 acggtacagg ccagacaatt attgtctggt atagtgcaac agcagaacaa tctgctgaag
     2881 gccattgagg cgcaacagca tatgttgcaa ctcacagtct ggggcatcaa gcagctccag
     2941 gcaagggtcc tggctgtgga aagataccta agggatcaac agctcctagg gatttggggc
     3001 tgctctggaa aactcatttg caacactgct gtgccttgga atactagttg gagtaataaa
     3061 tctcttaatg aaatttggaa taacatgacc tggttagagt gggaaagaga aattgacaat
     3121 tacacaggtt taatatacac cttacttgaa aaagcacaga accagcaaga aaagaatgaa
     3181 caagaattat tgaaattgga tagttgggca agtctgtgga attggtttag catatcaaac
     3241 tggctgtggt atataaggat attcataata gtagtaggag gcttagtagg tttaagaata
     3301 gtttttgctg tactttctat agtaaataga gttaggcagg gatattcacc attatcattg
     3361 cagacccgct cccaagcccc gaggggacca cccgacaggc ccggaggaat cgaagaagaa
     3421 ggtggagaga gagacagaga cagatccggt cgattagtca acggattctt gacacttatc
     3481 tgggacgacc tgaggagcct gtgcctcttc agctaccacc gcttgagaga cttactcttg
     3541 attgtaacga ggattgtgga acttctggga cacagggggt gggaagtcat caaatattgg
     3601 tggaatctcc tgcaatattg gagtcaggaa ctaaagaata gtgctattag cttgctaaat
     3661 gtcacagctg tagcagtagc tgaagggaca gatagaatta taacagtagt acaaagagct
     3721 tatagagctt ttcttcacat acctgtaaga ataagacaag gcttggaaag ggctctgcta
     3781 taagatgggt aacaagtggt caaaaggatg gcctgctgta agggaaagaa tgagacgagc
     3841 taagccagat gagccagcag cagaaggggt gggagcagta tctcgagacg gatccagcta
     3901 agggtcaaga caattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc
     3961 cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg
     4021 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     4081 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4141 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4201 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4261 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     4321 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4741 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4861 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4921 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4981 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5041 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5101 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     5161 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5221 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5281 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5341 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5401 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5461 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5521 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5581 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5641 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5701 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5761 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5821 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5881 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5941 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     6001 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6061 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6121 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     6181 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6241 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     6301 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6361 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6421 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6481 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6541 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6601 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6661 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6721 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6781 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6841 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6901 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6961 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7021 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7081 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7141 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7201 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7261 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7321 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7381 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7441 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7501 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7561 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7621 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7681 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7741 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7801 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7861 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7921 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7981 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8041 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8101 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8161 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8221 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8281 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8341 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8401 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8461 tgccacctga cgtc
//
