LOCUS       sequence-232470-        5990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of protein Rev from HIV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5990)
  AUTHORS   Blanco FJ, Hess S, Pannell LK, Rizzo NW, Tycko R
  TITLE     Solid-state NMR data support a helix-loop-helix structural model for
            the N-terminal half of HIV-1 Rev in fibrillar form.
  JOURNAL   J Mol Biol. 2001 Nov 2;313(4):845-59. doi: 10.1006/jmbi.2001.5067.
  PUBMED    11697908
REFERENCE   2  (bases 1 to 5990)
  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"; doi: "10.1006/jmbi.2001";
            journalName: "J Mol Biol"; date: "2001-11-2- 2"; volume: "313"; 
            issue: "4"; pages: "845-59"
COMMENT     Sequence Label: pET11d_Rev
FEATURES             Location/Qualifiers
     source          1..5990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..111
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        92..110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    111..135
                     /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)."
     RBS             150..172
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             401..427
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(582..600)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      596..643
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(818..823,824..840,841..846))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(873..891)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        959..1063
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1064..1132,1133..1924)
                     /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     complement(1282..1301)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2095..2683
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2584..2603
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2837..2854
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2869..3008
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2934..2953)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3094..3116
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3110..3301)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4613..5695)
                     /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     5657..5676
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5696..5773)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5979..5990,1..8)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
       61 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
      121 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
      181 ggctggtcgt tccggtgact ccgatgaaga cctgctcaaa gcagttcgtc tgatcaaatt
      241 cctgtaccag tctaacccgc caccgaaccc ggaaggcacc cgtcaggcgc gccgtaaccg
      301 tcgccgtcgc tggcgtgaac gtcagcgtca gatccactct attagcgagc gtatcctgtc
      361 cacctacctg ggtcgttccg ctgagccggt accgctgcag ctgccgccac tggaacgtct
      421 gactctggat tgcaacgaag actgcggtac ctctggcacc cagggtgttg gttctccgca
      481 gatcctggta gaaagcccga ctgttctgga aagcggcgct aaagagtaat aggatccggc
      541 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
      601 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
      661 atccggatat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
      721 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
      781 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
      841 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
      901 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
      961 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1021 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1081 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1141 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1201 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1261 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     1321 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     1381 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     1441 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1501 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1561 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     1621 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1681 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     1741 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     1801 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     1861 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1921 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1981 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2041 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2101 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2161 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2221 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2281 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2341 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2401 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2461 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2521 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2581 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2641 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2701 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     2761 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     2821 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     2881 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     2941 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     3001 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3061 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3121 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     3181 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     3241 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     3301 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     3361 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     3421 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     3481 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     3541 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     3601 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     3661 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     3721 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
     3781 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
     3841 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
     3901 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
     3961 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
     4021 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
     4081 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
     4141 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
     4201 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
     4261 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     4321 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     4381 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     4441 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     4501 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     4561 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     4621 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4681 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     4741 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     4801 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     4861 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     4921 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     4981 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     5041 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     5101 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     5161 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     5221 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     5281 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     5341 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     5401 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     5461 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     5521 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     5581 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     5641 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     5701 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     5761 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     5821 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     5881 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     5941 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
//
