LOCUS       sequence-273381-        8497 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 8497)
  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 8497)
  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: pHM089Env
FEATURES             Location/Qualifiers
     source          1..8497
                     /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             3995..4036
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4046..4063
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4086..4103)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4092..4316
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4362..4790
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4449..4468)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4659..4680
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4799..4819)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4804..5133
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4984..5119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5046..5065
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5200..5994
                     /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(5254..5273)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5864..5883
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6170..6291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6207..6226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6261..6280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6340..6356)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6340..6356)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6353..6375)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6364..6380
                     /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(6388..6394,6395..6412,6413..6418))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6433..6454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6571..6588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6742..7330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6822..6841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7501..8292,8293..8361))
                     /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     8124..8143
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8362..8466)
                     /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 gacctcctca gagcagtaag gatcatcaaa
     1021 atcctatacc aaagcagtaa gtaataaatc tatgtaatga cacctttggt aattcttgca
     1081 atagtaggac tggtagtagc gctaatctta gcaatagtag tgtggactat agtagctata
     1141 gaatatagaa aaatactaag ggaaagaaaa atagacaggt taattagaag aataaaagaa
     1201 agagaagagg acagtggaaa tgagagtgaa ggagacaccg atgaattggc caaacttgtg
     1261 gaaatggggg actttgatcc ttgggttggt gataatttgt agtgcctcag acaactcgtg
     1321 ggttacagtt tattatgggg ttcctgtgtg gaaagatgca gataccaccc tattttgtgc
     1381 ctcagatgct aaagcacatg acacagaagt gcacaatgtc tgggccacac atgcctgtgt
     1441 acccacagac cccaacccac aagaaatgta cttagaaaat gtaacagaaa attttaatat
     1501 gtggaaaaat aaaatggcag aacagatgca ggaggatgta atcagtttat gggatcaaag
     1561 tctaaagcca tgtgtaaagt taactcctct ctgtgttact ttacattgta ccaatgctaa
     1621 tttgaccaat gctaacaggg acaatgacac aaccaatgcc cctaacatag taggaaatat
     1681 gacagatgaa gtaaaaaact gtacttttaa tatgaccaca gaactaagag ataagaggca
     1741 aaatgtctat gcactttttt ataagcttga tatagtacaa ataggaagta ataataatag
     1801 tagtgagtat aggttaataa attgtaatac ctcagttatt aagcaggctt gtccaaagat
     1861 atcttttgat ccaattccta tacactattg tactccagct ggttatgcga ttttaaaatg
     1921 taatgataaa aagttcaatg ggacagggcc atgtaaaaat gtcagctcag tacaatgcac
     1981 acatggaatc aagccagtgg tatcaactca attgctgtta aatggcagtc tagcagaaga
     2041 agagataata atcagatctg aaaatctcac aaataatgcc aaaaccataa tagtgcacct
     2101 taataaatct gtagaaatca attgtaccag accctctaac aatacaagaa caagtataac
     2161 tatgggacca ggacaaatgt tctatagaac aggagacata ataggagata taaggaaagc
     2221 atattgtgag attaatggaa caaaatggaa tgaaacttta agccaagtag ctggaaaatt
     2281 aaaagagcac tttaataata agacaataat ctttcaacca cactcaggag gagatctaga
     2341 aattacaatg catcatttta attgtagagg ggaatttttc tattgcaata caacatcatt
     2401 gtttaatagt acttggacag gaaatgataa tagtacttgg aaagaaaata gcactatcac
     2461 gcttccatgc aggataaagc aaattataaa catgtggcag ggagtaggac aagcaatgta
     2521 tgctcctccc gtcagtggaa gcattaattg tgtatcaaat attacaggaa tcctattgac
     2581 aagagatggt ggtgtgcata atgggactaa tgagaccttc agacctggag gaggaaatat
     2641 aaaggacaat tggagaagtg aattatataa atataaagta gtacaaatag agccactagg
     2701 aatagcaccc accggggcaa agagaagagt ggtggagaga gaaaaaagag cagtgggaat
     2761 aggagctatg atctttgggt tcttaggagc agcaggaagc actatgggcg cggcgtcact
     2821 aacgctgacg gtacaggcca gacaattatt gtctggtata gtgcaacagc aaagcaattt
     2881 gctgagggct atagaggcgc agcagcatct gttgcaactc acagtctggg gcattaaaca
     2941 gctccaggca agagtcctgg ctgtggaaag atacctaaaa gatcaaaagt tcctaggact
     3001 ttggggctgc tctggaaaaa tcatctgcac cactgctgtg ccctggaact ccacttggag
     3061 taataaatct gttgacgaaa tttggaacaa catgacatgg atagaatggg agagagaaat
     3121 tagcaactac acaagccaaa tatatgagat acttacagaa tcacaaagcc agcaggataa
     3181 gaatgaaaag gatttgctgg aattggacaa atgggcaagt ctgtggaatt ggtttgacat
     3241 aacaaaatgg ctgtggtata taagaatatt tataatgata gtaggaggtt taataggttt
     3301 aagaataatt tttgctgtgc tttctatagt gaatagagtt aggcagggat actcaccttt
     3361 gtctctccag acccctttcc ctcagcagag ggaacccgac aggcccagag gaatcgaaga
     3421 agaaggtggc gaaacagaca gaggcagatc cgtgcgatta gtgagcgggt tcttagcgct
     3481 tgcgtgggac gacctgcgga gcctgtgcct cttcagctac caccgcttga gagacttcat
     3541 attgattgca gcgaggaccg tggaacttct gggacacagc agcctcaagg gactgaaacg
     3601 ggggtgggaa ggcctcaaat atctggggaa tcttctgttg tattggggtc gggaactaaa
     3661 gactagtgct atttctttgg tagatactat agcaatagta gtagcggggt ggacagatag
     3721 ggttatagaa gcagtacaaa caatttggag agccattctc cacataccta ggagaatcag
     3781 acagggctta gaaagggctt tgctataaca tgggaggcaa gtggtcaaag agtagcatag
     3841 tgggatggcc tcaggtcagg gaaagaataa ggcgaactcc tccagcagca gaaggggtag
     3901 gagcagtatc tcaagacgga tcaagggtca agacaattct gcagatatcc agcacagtgg
     3961 cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct
     4021 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     4081 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4141 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4201 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4261 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4321 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4381 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4441 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4501 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4561 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4621 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4681 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4741 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4801 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4861 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4921 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4981 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5041 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     5101 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5161 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5221 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5281 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5341 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5401 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5461 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5521 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5581 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5641 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5701 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5761 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5821 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5881 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5941 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6001 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6061 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6121 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6181 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6241 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6301 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6361 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6421 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6481 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6541 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6601 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6661 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6721 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6781 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6841 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6901 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6961 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7021 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7081 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7141 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7201 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7261 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7321 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7381 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7441 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7501 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7561 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7621 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7681 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7741 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7801 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7861 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7921 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7981 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8041 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8101 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8161 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8221 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8281 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8341 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8401 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8461 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
