LOCUS       sequence-305117-        6064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Gag P6 from HIV-1 HXB2 strain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  AUTHORS   Ramaraju B, Nelson SL, Zheng W, Ghirlando R, Deshmukh L
  TITLE     Quantitative NMR Study of Insulin-Degrading Enzyme Using 
            Amyloid-beta and HIV-1 p6 Elucidates Its Chaperone Activity.
  JOURNAL   Biochemistry. 2021 Aug 3. doi: 10.1021/acs.biochem.1c00342.
  PUBMED    34342986
REFERENCE   2  (bases 1 to 6064)
  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: 
            "Biochemistry. 2021 Aug 3. doi: 10.1021/acs.biochem.1c00342."
COMMENT     Sequence Label: pET-11a HIV-1 P6
FEATURES             Location/Qualifiers
     source          1..6064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(475..495)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(496..513)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(553..564)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(565..582)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     RBS             758..780
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    795..819
                     /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)."
     primer_bind     complement(819..838)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(820..838)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(922..941)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1147..1224
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1225..2307
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1244..1263)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3619..3810
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3804..3826)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3912..4051
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3967..3986
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4066..4083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4237..4825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4317..4336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4996..5787,5788..5856))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5619..5638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5857..5961)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6029..6047
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atccttactg gctgctcgga tcgttaccaa ataaagagcg
      361 taaagaggtc agcggataca gctctttatc gatcggttct tgtttctgcg gcggggtagt
      421 ggtttccacg ccactacgaa agctttcttc cggcggtgct gtcggttccg gacgactctg
      481 gaagtataaa ttctcatggt ggtggtgatg gtgttccaga aacagaccgc tatcctttaa
      541 attctcgcga atgcgacctt caatgctacc gcgcggaacc agttcggtca cggtgaaggt
      601 tttggtggcg tcatcatagg tccattcgcc atccacgcca ttatcattgg cgtactgttt
      661 aaacactttc tcggcggtgg cggcatcaac tgcttcggtg gtggtctcgc cttttaaagt
      721 cttgccattt aaagccagtt tatactgcat atgtatatct ccttcttaaa gttaaacaaa
      781 attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat
      841 ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg
      901 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
      961 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     1021 ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg
     1081 gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag
     1141 atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     1201 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     1261 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     1321 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     1381 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     1441 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     1501 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     1561 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     1621 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     1681 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     1741 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     1801 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     1861 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     1921 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     1981 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     2041 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     2101 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     2161 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     2221 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     2281 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca
     2341 ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     2401 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     2461 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
     2521 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     2581 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     2641 ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt
     2701 ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct
     2761 gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac
     2821 cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg cctcggcgag
     2881 cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc
     2941 gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc
     3001 gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat
     3061 gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg
     3121 cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg
     3181 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
     3241 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
     3301 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
     3361 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
     3421 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
     3481 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
     3541 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
     3601 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     3661 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     3721 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     3781 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     3841 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     3901 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     3961 actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca
     4021 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     4081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4681 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4741 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4801 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4861 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4921 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4981 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5041 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5101 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5161 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5221 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5281 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     5341 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5401 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5461 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5521 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5581 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     5641 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5701 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5761 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5821 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5881 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5941 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     6001 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     6061 agaa
//
