LOCUS       sequence-274211-        8446 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 8446)
  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 8446)
  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: pHM043Env
FEATURES             Location/Qualifiers
     source          1..8446
                     /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             3944..3985
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3995..4012
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4035..4052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4041..4265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4311..4739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4398..4417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4608..4629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4748..4768)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4753..5082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4933..5068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4995..5014
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5149..5943
                     /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(5203..5222)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5813..5832
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6119..6240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6156..6175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6210..6229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6289..6305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6289..6305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6302..6324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6313..6329
                     /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(6337..6343,6344..6361,6362..6367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6382..6403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6520..6537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6691..7279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6771..6790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7450..8241,8242..8310))
                     /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     8073..8092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8311..8415)
                     /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 gaactcctcg cagcagtaag agccatcaag
     1021 atcttatacc agagcagtaa gtatatgtaa tgacaccttt agaaattagt gcaatagcag
     1081 gactgatagt agcattaatc ttagcaatag tagtgtggac tatagtagct atacaaatta
     1141 agcaagtact aagagataga aaaatagaaa aattagttaa gagaataaga gagagagcag
     1201 aagacagtgg aaatgagagt gagggagaca cggatgaatt ggcaaaactg gtggaaatgg
     1261 gggactttga tccttgggtt ggggataatt tgtagtgcct cagacaactt gtgggttaca
     1321 gtttattatg gggttcctgt gtggaaagat gcagacacca ctctattttg tgcatcagat
     1381 gcccaagcac atcagacaga agtgcacaat gtctgggcca catatgcctg tgtacccaca
     1441 gaccccaacc cacaagaaat actcctggga aatgtaacag aaaattttaa catgtggaaa
     1501 aataacatgg tggagcagat gcaggaagat gtaatcagtt tatgggatca aagtctaaag
     1561 ccatgtgtaa agttaactcc cctctgcgtt actttaaatt gtaccgaggc taatctaagt
     1621 gccaacaaca catccaatat cactagcata gtaggaaata taacagagga agtaagaaac
     1681 tgctctttta atatgaccac agaactaaca gataagaggc agcaggtcca tgcacttttt
     1741 tataggcctg atatagtacc aattagaagc aatagtagtg agtataggtt aataaactgt
     1801 aatacttcag tcattaagca ggcttgtcca aaagtatcct ttgatccaat tcctatacat
     1861 tattgtactc cagctggtta tgcgatttta aaatgtaatg ataagaagtt caatgggaca
     1921 gggccatgta gaaatgtcag ctcaatacaa tgcacacatg gaattaagcc agtggtatca
     1981 actcagttgc tgttaaatgg tagtctagca gaagatgaga tagtaatcag aagtgaaaat
     2041 ctcacagaca atgccaagac cataatagtg catcttaata aatctgtaga catcaattgt
     2101 accagacctg gcaataatac aagaacaagt gtgtctatag gaccaggaca agtattctat
     2161 agacaaggag acataatagg agatataaca aaagcatatt gtgagattaa tggaacaaaa
     2221 tggaatgaaa ctttaacaca ggtaactaaa aaattaaagg agcactttaa taagacaata
     2281 ctctttcaac caccctcagg aggagatcta gaaattacaa tgcatcattt taattgtaga
     2341 ggggaatttt tctattgcaa tacaacaaaa ctgtttaatg gtacttgccc gcaaaatgga
     2401 acatgtaatg gcactatcat acttccatgt aagataaagc gaattgtaag attgtggcag
     2461 gaagtaggtc aagcaatgta tgctcctccc atcaagggaa gcattaggtg tgactcaaat
     2521 attacaggaa tactattgac aagagatggt ggtaatacta gtaataatga aaccttcaga
     2581 cctggaggag gaaatatgaa ggacaagtgg agaagtgaat tatataaata caaagtagta
     2641 caaattgaac cactaggaat agcacccacc agggcaaaaa gaagagtggt ggagagagaa
     2701 aagagagcag tgggaatagg agctatgatc tttgggttct taggagcagc aggaagcact
     2761 atgggcgcag cgtcaataac gctgacggta catgccagac aattattgtc cggtatagtg
     2821 caacagcaaa gcaatttgct gagggctata gaggcgcagc agcatctgtt gcaactcaca
     2881 gtctggggca ttaaacagct ccaggcaaga gtcctggctg tggaaagata cctaaaagat
     2941 caaaagttcc taggactttg gggctgctct gggaaaatca tctgcaccac tgctgtgcct
     3001 tggaacacca cttggagtaa taaatctttt gaagagattt gggacaacat gacatggata
     3061 gaatgggaga gagaaattag caattacaca aaccaaatat atgagatact tacagaatcg
     3121 cagaaccagc aggataaaaa tgaaaaggat ctgttagaat tggataagtg ggcaagtctg
     3181 tggaattggt ttgacataac aaggtggctg tggtatataa aaatgtttat aataatagta
     3241 ggaggtttaa taggtttaag aataattttt gctgtgcttt ctatagtaaa tagagttagg
     3301 cagggatact cacctttgtc tttccagacc ccttcccatc atcagaggga acccgacagg
     3361 cccgaaggaa tcgaagaagg aggtggcgag caaggcagag acaaatccgt gcgattagtg
     3421 agcggatttt taccacttat ctgggacgat ctgcggagcc tgtgcctctt cagctaccac
     3481 cgcttgagag acttcatctt gattgcaacg aggactgtgg aacttctggg acacagcagt
     3541 ctcaagggac tgagacgggg gtgggaaggc ctcaaatatc tggggaatct tctcctatat
     3601 tggggtcagg aactaaaaac tagtgctatt tctttgtttg atgctgcagc aatagcagta
     3661 gcggggtgga cagatagggt tatagaagta gcacaaagag cttggagagc cattctccac
     3721 atacctagaa ggatcagaca gggctttgaa agggctttgc tataatatgg gaaacaagtg
     3781 gtcaaaaaca tggcctcggg tcagggaaac aatagaaaaa acttctccag cagcagatgg
     3841 agtaggagca gtatctcaag aggatccagc taagggtcaa gacaattctg cagatatcca
     3901 gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
     3961 ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta
     4021 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4081 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4141 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4201 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4261 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4321 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4381 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4441 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4501 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4561 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4621 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4681 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4741 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4801 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4861 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4921 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4981 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5041 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5101 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5161 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5221 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5281 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5341 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5401 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5461 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5521 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5581 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5641 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5701 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5761 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5821 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5881 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5941 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6001 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6061 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6121 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6181 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6241 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6301 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6361 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6421 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6481 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7261 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7321 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7381 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7441 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7501 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7561 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7621 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7681 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7741 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7801 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7861 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7921 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7981 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8041 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8101 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8161 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8221 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8281 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8341 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8401 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
