LOCUS       sequence-274146-        8519 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 8519)
  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 8519)
  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: pHM149Env
FEATURES             Location/Qualifiers
     source          1..8519
                     /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             4017..4058
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4068..4085
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4108..4125)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4114..4338
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4384..4812
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4471..4490)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4681..4702
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4821..4841)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4826..5155
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5006..5141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5068..5087
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5222..6016
                     /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(5276..5295)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5886..5905
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6192..6313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6229..6248)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6283..6302
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6362..6378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6362..6378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6375..6397)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6386..6402
                     /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(6410..6416,6417..6434,6435..6440))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6455..6476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6593..6610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6764..7352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6844..6863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7523..8314,8315..8383))
                     /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     8146..8165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8384..8488)
                     /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 gaactcctca gggcagtaag gatcgtcaac
     1021 atcctatacc agagcagtaa gtatatgtaa tgacaccttt agaaattagt gcaatagcag
     1081 gactgataat agcattgatc ttagcaatag tagtgtggac tatagtaggt atacaactta
     1141 agaaaatact aaagcaaaga aaaatagaca ggttaattga gagaataaga gaaagagaag
     1201 aagacagtgg aaatgaaagt gaaggagaca cagatgaatt ggccaaactt gtggaaatgg
     1261 gggaccttga tccttgggtt ggtgatattt tgtagtgctg cagacaactt gtgggttaca
     1321 gtttattatg gggttcctgt gtggagagat gcagagacta ctctattttg tgcatcagat
     1381 gccaaagcac atgagacaga agtgcacaat gtctgggcca cacatgcttg tgtacccaca
     1441 gaccccaacc cacaagaaat gctcctggaa aatgtaacag aaaattttaa catgtggaaa
     1501 aataaaatgg tagagcaaat gcaggaggat gtaatcagtt tatgggatca aagtttacag
     1561 ccatgtgtaa agttaactcc tctctgcgtc actttaaatt gtaccgatgc taagttaagc
     1621 aataccaaca gcagtaacca agagggccct agtatgcaag agataggaaa tataacagat
     1681 gaagtaagaa actgttcttt taatacgacc acagaactaa gagataggca gcagaaggtc
     1741 catgcacttt tttataggct tgatatagta aaagttgata acaataactc aagtgaatat
     1801 aggttaataa attgtaatac ttcagtcatt aagcaggctt gcccaaaggt atcctttgat
     1861 ccaattccta tacattattg tactccagct ggttatgcgc ttttgaagtg taatgataag
     1921 aaattcaatg ggacagggcc atgtaaaaat gtcagctcag tacaatgcac acatggaatc
     1981 aagccagtgg tatcaactca attgctgtta aatggcagtc tagcagaagg agaggtaatg
     2041 atcagatctg aaaatttcac aaacaatgcc aaaatcataa tagtgcatct taatgaatct
     2101 gtaataatca attgtaccag accctccaac aatacaagaa caagtataaa tatgggacca
     2161 ggacaagtac tatatagaac aggagacata ataggagata taagaagagc atattgtaag
     2221 gttaatagaa caagatggta tgaaacttta aaacaggtaa ctggaaaatt gaaagagcac
     2281 tttaatagga caataatctt tcaaccaccc tcaggaggag atatagaaat aacaacgcat
     2341 cattttaatt gtagagggga atttttctat tgcaatacat caaaactgtt taatagtact
     2401 tggggttggg ggcaaaatat aaccatggaa aagtgtaata acactatgtg taatgaaact
     2461 gatatcatac tcccatgcag gataaagcaa attataaaca tgtggcagag agcaggacag
     2521 gcaatgtatg ctcctcccat caagggacac attagttgtg tatcaaatat tacaggaata
     2581 atattgacaa gagatggtgg tgttaataat acaagcaacg agaccttcag acctggagga
     2641 ggagatataa aggacaattg gagaagtgaa ttatataaat ataaagtagt acaaattgaa
     2701 ccactaggaa tagcaccatc caaggcacag agaagagtgg tggacagaca aaaaagagca
     2761 gtgggaatag gagctatgat ctttgggttc ttaggagcag ccgggagcac tatgggcgca
     2821 gcgtcactga cgctgacggt acaggccaga caattattgt ctggtatagt gcaacagcaa
     2881 agcaatttgc tgagggctat tgaggcgcag cagcatctgt tgcaactcac agtctggggc
     2941 attaaacagc tccaggcgag agtcctggct gtggaaagat acctaaagga tcaaagactc
     3001 ctaggacttt ggggctgctc tggaagaacc atctgcacca ctaatgtgcc ttggaactcc
     3061 acttggagta ataaatctta tgatgagatt tggggcaaca tgacatggat agaatgggag
     3121 agagaaatta gtaattacac agaccaaata tatggcatac ttacagaatc gcaaaaccag
     3181 caggaaaaga atgaaaagga tttgttagaa ttggataagt gggcaagtct gtggaattgg
     3241 tttgacataa caaactggct gtggtatata aaaatattta taatgatagt aggaggctta
     3301 ataggtttaa gaataatttt tgctatgctt tctatagtaa atagggttag gcagggatac
     3361 tcacctttgt ctttccagat cccatcccat catcagaggg aacccgacag gcccggagga
     3421 atcgaagaag aaggtggcga gcaaggcaga gacagatcag tgcgcttagt gagcggattc
     3481 ttagcactgt tttgggacga cctacggagc ctgtgcctct tcagcttcca cctattgaga
     3541 gactccacct tgattgcagc gaggactttg gaacttctgg gacgcagcag tctcaaggga
     3601 ctgagacagg ggtgggaagg cctcaaatat ctggggaatc tcctgatata ttggatccag
     3661 gaactaaaaa ctagtgctgt ttctttgctt gatgctgtag cagtagcagt agcagggtgg
     3721 acagataggg ttatagaagt agcacaaaga gctgggagag ccattctcaa catacctaga
     3781 agaatcagac agggtttaga aaggtcttta ctataacatg ggaaacaagt ggtcaaaaca
     3841 atggcctcag gtcagagaaa gaatgaggca gactcctcta aggcaaactc aaactcctcc
     3901 agcagcagaa ggagtaggag cagtatctca agacggatcc agctaagggt caagacaatt
     3961 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta
     4021 agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc
     4081 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     4141 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     4201 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     4261 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     4321 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     4381 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4441 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4501 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4561 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4621 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4681 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4741 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4801 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4861 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4921 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4981 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5041 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     5101 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5161 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     5221 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5281 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5341 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5401 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5461 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5521 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5581 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5641 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5701 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5761 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5821 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5881 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5941 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6001 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     6061 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6121 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6181 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6241 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6301 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     6361 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6421 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6481 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6541 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6601 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6661 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6721 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6781 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6841 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6901 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6961 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7021 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7081 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7141 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7201 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7261 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7321 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7381 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7441 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7501 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7561 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7621 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7681 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7741 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7801 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7861 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7921 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7981 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8041 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8101 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8161 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8221 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8281 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8341 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8401 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8461 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
