LOCUS       sequence-234151-        8142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8142)
  AUTHORS   Lemaire PA, Lary J, Cole JL
  TITLE     Mechanism of PKR activation: dimerization and kinase activation in 
            the absence of double-stranded RNA.
  JOURNAL   J Mol Biol. 2005 Jan 7;345(1):81-90.
  PUBMED    15567412
REFERENCE   2  (bases 1 to 8142)
  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: "J Mol 
            Biol."; date: "2005-01-7"; volume: "345(1)"; pages: "81-90"
COMMENT     Sequence Label: pPET-PKR/PPase
FEATURES             Location/Qualifiers
     source          1..8142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             1848..1870
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2573..2590
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2643..2661)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2657..2704
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2879..2884,2885..2901,2902..2907))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2934..2952)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3020..3124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3125..3193,3194..3985)
                     /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(3343..3362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4156..4744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4645..4664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4898..4915
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4930..5069
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4995..5014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5155..5177
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5171..5362)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6674..7756)
                     /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     7718..7737
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7757..7834)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     8040..8059
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggctggtgat ctttcagcag gtttcttcat
      121 ggaggaactt aatacatacc gtcagaagca gggagtagta cttaaatatc aagaactgcc
      181 taattcagga cctccacatg ataggaggtt tacatttcaa gttataatag atggaagaga
      241 atttccagaa ggtgaaggta gatcaaagaa ggaagcaaaa aatgccgcag ccaaattagc
      301 tgttgagata cttaataagg aaaagaaggc agttagtcct ttattattga caacaacgaa
      361 ttcttcagaa ggattatcca tggggaacta cataggcctt atcaatagaa ttgcccagaa
      421 gaaaagacta actgtaaatt atgaacagtg tgcatcgggg gtgcatgggc cagaaggatt
      481 tcattataaa tgcaaaatgg gacagaaaga atatagtatt ggtacaggtt ctactaaaca
      541 ggaagcaaaa caattggccg ctaaacttgc atatcttcag atattatcag aagaaacctc
      601 agtgaaatct gactacctgt cctctggttc ttttgctact acgtgtgagt cccaaagcaa
      661 ctctttagtg accagcacac tcgcttctga atcatcatct gaaggtgact tctcagcaga
      721 tacatcagag ataaattcta acagtgacag tttaaacagt tcttcgttgc ttatgaatgg
      781 tctcagaaat aatcaaagga aggcaaaaag atctttggca cccagatttg accttcctga
      841 catgaaagaa acaaagtata ctgtggacaa gaggtttggc atggatttta aagaaataga
      901 attaattggc tcaggtggat ttggccaagt tttcaaagca aaacacagaa ttgacggaaa
      961 gacttacgtt attaaacgtg ttaaatataa taacgagaag gcggagcgtg aagtaaaagc
     1021 attggcaaaa cttgatcatg taaatattgt tcactacaat ggctgttggg atggatttga
     1081 ttatgatcct gagaccagtg atgattctct tgagagcagt gattatgatc ctgagaacag
     1141 caaaaatagt tcaaggtcaa agactaagtg ccttttcatc caaatggaat tctgtgataa
     1201 agggaccttg gaacaatgga ttgaaaaaag aagaggcgag aaactagaca aagttttggc
     1261 tttggaactc tttgaacaaa taacaaaagg ggtggattat atacattcaa aaaaattaat
     1321 tcatagagat cttaagccaa gtaatatatt cttagtagat acaaaacaag taaagattgg
     1381 agactttgga cttgtaacat ctctgaaaaa tgatggaaag cgaacaagga gtaagggaac
     1441 tttgcgatac atgagcccag aacagatttc ttcgcaagac tatggaaagg aagtggacct
     1501 ctacgctttg gggctaattc ttgctgaact tcttcatgta tgtgacactg cttttgaaac
     1561 atcaaagttt ttcacagacc tacgggatgg catcatctca gatatatttg ataaaaaaga
     1621 aaaaactctt ctacagaaat tactctcaaa gaaacctgag gatcgaccta acacatctga
     1681 aatactaagg accttgactg tgtggaagaa aagcccagag aaaaatgaac gacacacatg
     1741 ttagagccct tctgaaggat ccccgggaat tcatcgtgac tgactgacga tctgcctcaa
     1801 ttcgagctcc gtcgacaagc ttgcggccgc actcgactag aaataatttt gtttaacttt
     1861 aagaaggaga tatacatatg cgctattacg aaaaaattga tggcagcaaa taccgaaata
     1921 tttgggtagt tggcgatctg cacggatgct acacgaacct gatgaacaaa ctggatacga
     1981 ttggattcga caacaaaaaa gacctgctta tctcggtggg cgatttggtt gatcgtggtg
     2041 cagagaacgt tgaatgcctg gaattaatca cattcccctg gttcagagct gtacgtggaa
     2101 accatgagca aatgatgatt gatggcttat cagagcgtgg aaacgttaat cactggctgc
     2161 ttaatggcgg tggctggttc tttaatctcg attacgacaa agaaattctg gctaaagctc
     2221 ttgcccataa agcagatgaa cttccgttaa tcatcgaact ggtgagcaaa gataaaaaat
     2281 atgttatctg ccacgccgat tatccctttg acgaatacga gtttggaaag ccagttgatc
     2341 atcagcaggt aatctggaac cgcgaacgaa tcagcaactc acaaaacggg atcgtgaaag
     2401 aaatcaaagg cgcggacacg ttcatctttg gtcatacgcc agcagtgaaa ccactcaagt
     2461 ttgccaacca aatgtatatc gataccggcg cagtgttctg cggaaaccta acattgattc
     2521 aggtacaggg agaaggcgca tgagaattcg agctccgtcg acaagcttcg agcaccacca
     2581 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     2641 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     2701 tttgctgaaa ggaggaacta tatccggata tcccgcaaga ggcccggcag taccggcata
     2761 accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg
     2821 ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat
     2881 taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt
     2941 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3001 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3061 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3121 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3181 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3241 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3301 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3361 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3421 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3481 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3541 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3601 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3661 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3721 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3781 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3841 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3901 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3961 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4021 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4081 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4141 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4201 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4261 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     4321 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4381 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4441 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4501 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4561 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4621 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4681 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4741 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4801 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4861 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4921 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     4981 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     5041 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     5101 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5161 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     5221 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     5281 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     5341 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     5401 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     5461 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     5521 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     5581 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     5641 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     5701 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     5761 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     5821 gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg
     5881 cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga
     5941 attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat
     6001 tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta
     6061 tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat
     6121 cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc
     6181 ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg
     6241 catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt
     6301 cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg
     6361 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     6421 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     6481 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     6541 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     6601 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     6661 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6721 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     6781 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     6841 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     6901 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     6961 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     7021 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     7081 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     7141 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     7201 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     7261 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     7321 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     7381 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     7441 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     7501 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     7561 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     7621 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     7681 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     7741 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     7801 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     7861 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     7921 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     7981 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     8041 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     8101 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa at
//
