LOCUS       sequence-472260-        6242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Rnr nuclease domain (residues 649 to 1929) as a 
            GST-fusion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6242)
  AUTHORS   Baijal K, Kore B, Abramchuk I, Denoncourt A, Han S, Simms A, 
            Dagenais A, Long AR, Rudner AD, Lavallee-Adam M, Gray MJ, Downey M
  TITLE     Identification of polyphosphate-binding proteins in E. coli uncovers
            targets involved in translation control and ribosome biogenesis
  JOURNAL   bioRxiv 2025.02.12.637445
REFERENCE   2  (bases 1 to 6242)
  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: "bioRxiv 
            2025.02.12.637445"
COMMENT     Sequence Label: pRnr-ND
FEATURES             Location/Qualifiers
     source          1..6242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        833..910
                     /gene="lacI"
                     /label=lacI promoter
     CDS             911..1993
                     /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(930..949)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2042..2047,2048..2065,2066..2072)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2080..2096
                     /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     2104..2120
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2104..2120
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2116..2643
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2136..2153)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2136..2152)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2145..2167)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2745..2750,2751..2766,2767..2773)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2781..2797
                     /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)."
     CDS             2820..3473
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3431..3453
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3480..3497
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4854..4876)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5021..5039)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5107..5211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5212..5280,5281..6072)
                     /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(5430..5449)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat
      841 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
      901 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
      961 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     1021 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     1081 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     1141 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     1201 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
     1261 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     1321 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     1381 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     1441 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     1501 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     1561 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     1621 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     1681 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     1741 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     1801 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     1861 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     1921 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     1981 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg
     2041 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     2101 acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt
     2161 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2221 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2281 aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg
     2341 gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt
     2401 tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt
     2461 cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta
     2521 caggaaggcc agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg
     2581 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     2641 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     2701 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
     2761 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagtattca
     2821 tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
     2881 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
     2941 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
     3001 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
     3061 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
     3121 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
     3181 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
     3241 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
     3301 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
     3361 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
     3421 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc
     3481 tggttccgcg tggatccccg gaattcatga atatgggcac cggcatggcg gttgatatcg
     3541 ctctgcgtac ccatgaaatt ccgtacatct ggccgcaggc tgttgagcaa caggttgccg
     3601 ggctgaaaga agaagtgccg gaagaagcaa aagcgggccg tgttgatctg cgcgatttac
     3661 cgctggtcac cattgatggc gaagacgccc gtgactttga cgatgcagtt tactgcgaga
     3721 aaaaacgcgg cggcggctgg cgtttatggg tcgcgattgc cgacgtcagc tactatgtgc
     3781 gtccgtcaac gccgctggac agagaagcgc gtaaccgtgg cacgtcggtg tacttccctt
     3841 cgcaggttat cccgatgctg ccggaagtgc tctctaacgg cctgtgttcg ctcaacccgc
     3901 aggtagaccg cctgtgtatg gtgtgcgaga tgacggtttc gtcgaaaggc cgcctgacgg
     3961 gctacaaatt ctacgaagcg gtgatgagct ctcacgcgcg tctgacctac accaaagtct
     4021 ggcatattct gcagggcgat caggatctgc gcgagcagta cgccccgctg gttaagcatc
     4081 tcgaagagtt gcataacctc tataaagtgc tggataaagc ccgtgaagaa cgcggtggga
     4141 tctcatttga gagcgaagaa gcgaagttca ttttcaacgc tgaacgccgt attgaacgta
     4201 tcgaacagac ccagcgtaac gacgcgcaca aattaattga agagtgcatg attctggcga
     4261 atatctcggc ggcgcgtttc gttgagaaag cgaaagaacc ggcactgttc cgtattcacg
     4321 acaagccgag caccgaagcg attacctctt tccgttcagt gctggcggag ctggggctgg
     4381 aactgccggg cggtaacaag ccggaaccgc gtgactacgc ggagctgctg gagtcggttg
     4441 ccgatcgtcc tgatgcagaa atgctgcaaa ccatgctgct gcgctcgatg aaacaggcga
     4501 tttacgatcc agaaaaccgt ggtcactttg gcctggcatt gcagtcctat gcgcacttta
     4561 cttcgccgat tcgtcgttat ccagacctga cgctgcaccg cgccattaaa tatctgctgg
     4621 cgaaagagca ggggcatcag ggcaacacca ctgaaaccgg cggctaccat tattcgatgg
     4681 aagagatgct gcaactgggt cagcactgtt cgatggcgga acgtcgtgcc gacgaagcaa
     4741 cgcgcgatgt ggctgactgg ctgaagtgtg acttcatgct cgaccaggta tgagcggccg
     4801 catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     4861 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4921 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     4981 cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt
     5041 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5101 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5161 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5221 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5281 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5341 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5401 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     5461 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5521 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5581 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5641 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5701 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     5761 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5821 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5881 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5941 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6001 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6061 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6121 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6181 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6241 tc
//
