LOCUS       sequence-468400-        5779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of the yeast PPX1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5779)
  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 5779)
  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: pScPPX1
FEATURES             Location/Qualifiers
     source          1..5779
                     /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"
     misc_feature    775..915
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(841..860)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1001..1023
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1161..1180
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1360..2238)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2265..2549
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2472..2491
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3826..3843)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3996..4082
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4174..4201
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4220..4311
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4312..4380,4381..5172)
                     /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(4530..4549)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5214..5669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5301..5320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5511..5532
                     /label=F1ori-F
                     /note="F1 origin, forward 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 cgcatatggt gcactctcag
      841 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
      901 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
      961 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     1021 aggttttcac cgtcatcacc gaaacgcgcg aggcagaagg agatggcgcc caacagtccc
     1081 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     1141 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     1201 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tctgctcatg tttgacagct
     1261 tatcatcgat gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg
     1321 ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac
     1381 atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt
     1441 tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc
     1501 gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg
     1561 ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga
     1621 caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct
     1681 gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat
     1741 cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata
     1801 gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg
     1861 gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca
     1921 ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc
     1981 cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg
     2041 gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag
     2101 aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc
     2161 aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc
     2221 attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc
     2281 atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc
     2341 ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac
     2401 gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt
     2461 cacactttgc tatgccatag catttttatc cataagatta gcggatccta cctgacgctt
     2521 tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagcg aattcatgtc
     2581 gcctttgaga aagacggttc ctgaattttt ggcacactta aaatcactgc caatttcaaa
     2641 aattgcaagc aacgatgtgt taacaatatg tgttggtaac gagtcagcag atatggactc
     2701 aattgctagt gcaatcactt attcgtactg ccaatacata tataatgaag gtacttactc
     2761 ggaggagaaa aagaaaggaa gctttattgt cccaattatc gacattccta gagaagacct
     2821 cagtttaaga agagacgtaa tgtatgtcct agaaaaactg aaaattaagg aggaggaatt
     2881 attcttcatt gaagatttaa agagcttgaa gcaaaatgtc tcccagggta ctgaattaaa
     2941 ctcttacttg gtagataata acgatacacc aaagaatttg aaaaattata tagataacgt
     3001 cgttggcatt atagaccacc attttgactt gcaaaaacat ttggatgctg aacctcggat
     3061 tgtaaaagtg tccggcagtt gctcatcgct ggtttttaac tactggtatg aaaaattgca
     3121 aggtgaccgt gaagtggtga tgaacattgc accacttttg atgggggcca tcttaataga
     3181 cacttcaaat atgaggcgca aagtcgagga aagtgataaa ttagctatcg agagatgcca
     3241 agctgttctt agtggtgcgg ttaatgaagt gtctgcgcaa ggtttagagg acagcagtga
     3301 gttttataaa gagataaaat caagaaagaa cgatattaaa ggattttcgg taagcgatat
     3361 tctaaagaag gactacaaac aattcaattt ccaaggaaag ggacacaaag ggttagagat
     3421 tggtctttca tcaatagtaa aaagaatgtc ttggctattc aatgaacacg gtggtgaagc
     3481 agatttcgtc aaccaatgca gaagatttca ggcggagagg gggctcgatg tattggttct
     3541 gttgacttca tggaggaaag ctggtgattc acacagagaa ttggtcatat tgggagactc
     3601 taacgtggta cgtgaactca ttgaaagggt tagcgacaag ctccaacttc aattatttgg
     3661 gggcaatctt gatggaggtg tggcgatgtt taagcaactg aacgtcgagg ccaccagaaa
     3721 gcaagtcgtc ccctatttag aggaagcgta ctcaaacctg gaagagtgag tcgacctgca
     3781 ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     3841 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     3901 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     3961 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     4021 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     4081 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     4141 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     4201 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4261 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4321 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4381 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4441 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4501 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     4561 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4621 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4681 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4741 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4801 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     4861 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4921 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4981 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5041 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5101 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5161 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc
     5221 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5281 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5341 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5401 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     5461 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5521 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     5581 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5641 ttttaacaaa atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata
     5701 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5761 aaaagatcaa aggatcttc
//
