LOCUS       sequence-273386-        8504 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 8504)
  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 8504)
  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: pHM033Env
FEATURES             Location/Qualifiers
     source          1..8504
                     /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             4002..4043
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4053..4070
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4093..4110)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4099..4323
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4369..4797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4456..4475)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4666..4687
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4806..4826)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4811..5140
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4991..5126
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5053..5072
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5207..6001
                     /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(5261..5280)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5871..5890
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6177..6298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6214..6233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6268..6287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6347..6363)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6347..6363)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6360..6382)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6371..6387
                     /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(6395..6401,6402..6419,6420..6425))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6440..6461
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6578..6595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6749..7337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6829..6848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7508..8299,8300..8368))
                     /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     8131..8150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8369..8473)
                     /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 caccgacgaa gaactcctca gagcagtaag gatcatcaaa
     1021 atcctaaacc agagcagtaa gtaaatgcaa tgacaccttt gcaaattagt gcaatagcag
     1081 gattgatagt agcgctaatc gcagcaatag tagtgtggac tatagtagct atagaagtta
     1141 agaaattata taggcaaagc aaaatagaca ggttagttaa aagaataaga gaaagagcag
     1201 aagacagtgg aaatgagagt gaaggggaca cagatgaatt ggccaaactt gtggaaatgg
     1261 gggactttga tccttgggtt ggtgataatt tgtagtgcct cagacaagtt gtgggttaca
     1321 gtttattatg gagttcctgt gtggaaagat gcagatacca ccctattttg tgcatcagat
     1381 gctaaagcac aggagacaga agtgcacaat atctgggcca cacatgcctg tgtacccaca
     1441 gaccccaacc cacaagaaat acacctgaaa aatgtaacag aaaattttaa catgtggaaa
     1501 aataacatgg tagagcagat gcaggaggat gtaatcagtt tatgggatca aagtctaaag
     1561 ccatgtgtaa agttaactcc tctctgtgtt actttaaatt gtagcattgc tacttttccc
     1621 aatgacacta tgtccaggga caatagcagt atcaatgacg ataccatagg aaatctaaca
     1681 gaggagttaa aaaactgttc ttttagtata accacagaac taagagataa gaagcaaaag
     1741 gcccatgcac ttttttataa gcttgatata gtaccaattg aagatagtgg taatagtagt
     1801 acctataggt taataaattg taatacttca gtcattaagc aggcttgtcc aaaggtatcc
     1861 tttgatccaa ttcccataca ttattgtact ccagctggtt atgcgatttt caagtgtaat
     1921 gataagaatt tcaatgggac agggccatgt aaaaatgtca gctcagtaca atgcacacat
     1981 ggaattaagc cggtggtatc aactcaattg ctgttaaatg gcagtctagc agaagaagag
     2041 ataataatca gatctgaaaa cctcacaaat aatgccaaaa ccataatagt gcacctcaat
     2101 gactctgtag aaatcacttg taccagaccc tccaacaata caagaacaag tttaactata
     2161 ggaccaggac aagtgttcta tagaacagga gacataatag gagatataag aagggcacat
     2221 tgtgtgatta atggagcaaa atggaatgaa actttaaaac aggtagctaa aaaattaaga
     2281 gaaagtttta ataagacaat aatctttcaa ccacccgtag gaggagatcc agaaattaca
     2341 atgcttcatt ttaattgcag aggggaattt ttctattgca atacaacaaa actgtttaat
     2401 agtacttggg gagaaaatga aaacataacc acgggggggt ctaataacac tcacactgtc
     2461 attccatgca ggataaagca aattataaac atgtggcagg gaacaggaca agcaatgtat
     2521 gctcctccca tcaggggaaa cattacttgt gtatcaaaca ttacaggaat actattgaca
     2581 agagatggtg gtgataataa tagtattaac gagaccttca gacctggagg aggaaatata
     2641 aaggacaatt ggagaagtga attatataaa tataaagtag tacaaattga accactagga
     2701 gtagcaccct ccaaggcaag gagaagagtg gtggagagag aaaaaagagc agttggaata
     2761 ggagctatga tctttgggtt cttaggagca gcaggaagca ctatgggcgc ggcgtcgcta
     2821 acgctgacgg tacaggccag acaattattg tctggtatag tgcaacagca aagcaatttg
     2881 ctgagggcta tagaggcgca gcagcatatg ttgcaactca cagtctgggg cattaaacag
     2941 ctccaggcaa gagtcctggc tgtggaaaga tacctaaagg atcaaaagtt cctaggactt
     3001 tggggctgct ctggaaaaat catctgcacc actaatgtgc cttggaactc cacttggagt
     3061 aataaatctt atgcagatat ttggaacaat atgacatgga cagaatggga gagagaaatt
     3121 agcaattaca caaaccaaat atatgaattg cttacagaat cgcagaacca gcaggacaag
     3181 aatgaaaagg atttgttgga attggataaa tgggcaagct tgtggacttg gtttgacata
     3241 acacagtggc tgtggtatat aaaaatattt ataatgataa taggaggctt aataggttta
     3301 agaataattt ttgctgtgct ttctatagta aatagagtta ggcaggggta ctcacctttg
     3361 tctttccaga cccctaccca tcatcagagg gaacaacccg acagactcga aagaatcgaa
     3421 gaagaaggtg gcgagcaagg cagagacaga tccgtgcgct tagtgagcgg attcttggct
     3481 ctgttctggg acgatctacg gagcctgtgc ctcttcagct accgccgctt gagagacttc
     3541 agcttgattg taacgaggac tgtggaactt ctgggacaca gcagtctcaa gggactgaga
     3601 cgggggtggg aaggcctcaa atatctgggg aatcttctgt tgtattgggg tcaggaacta
     3661 aagattagtg ctgtttcttt gtttgatgct atagccataa gagtagcggg gtggacagat
     3721 aggattatag aagtagcaca aagagcttgg agagctattc tccacatacc tagaagaatc
     3781 aggcagggct ttgaaagggc tttgttataa catgggaaac aagtggtcaa aaaatagcat
     3841 agtgggatgg cctcagatca gggaaagaat aaggagaact cctccagcag cagaaggcgt
     3901 aggagcagta tctcaagacg gatccagcta agggtcaaga caattctgca gatatccagc
     3961 acagtggcgg ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc
     4021 ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa
     4081 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     4141 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     4201 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     4261 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     4321 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     4381 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4441 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4501 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4561 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4621 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4681 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4741 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4801 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4861 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4921 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4981 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5041 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     5101 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     5161 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     5221 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     5281 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5341 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     5401 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5461 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5521 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5581 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5641 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5701 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5761 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5821 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5881 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5941 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     6001 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6061 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6121 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6181 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6241 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6301 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6361 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6421 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6481 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6541 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6601 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6661 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6721 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6781 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6841 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6901 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6961 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7021 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7081 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7141 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7201 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7261 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7321 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7381 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7441 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7501 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7561 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7621 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7681 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7741 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7801 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7861 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7921 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7981 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8041 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8101 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8161 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8221 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8281 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8341 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8401 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8461 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
