LOCUS       sequence-219996-        6891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His-tagged yeast exopolyphosphatase for purification from 
            E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6891)
  AUTHORS   Gray MJ, Wholey WY, Wagner NO, Cremers CM, Mueller-Schickert A, Hock
            NT, Krieger AG, Smith EM, Bender RA, Bardwell JC, Jakob U
  TITLE     Polyphosphate is a primordial chaperone.
  JOURNAL   Mol Cell. 2014 Mar 6;53(5):689-99. doi: 
            10.1016/j.molcel.2014.01.012. Epub 2014 Feb 20.
  PUBMED    24560923
REFERENCE   2  (bases 1 to 6891)
  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: 
            "10.1016/j.molcel.2014.01.012"; journalName: "Mol Cell"; date: 
            "2014-03-6- 6"; volume: "53"; issue: "5"; pages: "689-99"
COMMENT     Sequence Label: pScPPX2
FEATURES             Location/Qualifiers
     source          1..6891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..80
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        61..79
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    80..104
                     /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             119..141
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             160..177
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             187..204
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1452..1470)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1466..1513
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1688..1693,1694..1710,1711..1716))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1743..1761)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1829..1933
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1934..2002,2003..2794)
                     /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(2152..2171)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2965..3553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3454..3473
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3707..3724
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3739..3878
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3804..3823)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3964..3986
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3980..4171)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5483..6565)
                     /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     6527..6546
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6566..6643)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6849..6868
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
       61 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
      121 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      181 agcggcctgg tgccgcgcgg cagccatatg tcgcctttga gaaagacggt tcctgaattt
      241 ttggcacact taaaatcact gccaatttca aaaattgcaa gcaacgatgt gttaacaata
      301 tgtgttggta acgagtcagc agatatggac tcaattgcta gtgcaatcac ttattcgtac
      361 tgccaataca tatataatga aggtacttac tcggaggaga aaaagaaagg aagctttatt
      421 gtcccaatta tcgacattcc tagagaagac ctcagtttaa gaagagacgt aatgtatgtc
      481 ctagaaaaac tgaaaattaa ggaggaggaa ttattcttca ttgaagattt aaagagcttg
      541 aagcaaaatg tctcccaggg tactgaatta aactcttact tggtagataa taacgataca
      601 ccaaagaatt tgaaaaatta tatagataac gtcgttggca ttatagacca ccattttgac
      661 ttgcaaaaac atttggatgc tgaacctcgg attgtaaaag tgtccggcag ttgctcatcg
      721 ctggttttta actactggta tgaaaaattg caaggtgacc gtgaagtggt gatgaacatt
      781 gcaccacttt tgatgggggc catcttaata gacacttcaa atatgaggcg caaagtcgag
      841 gaaagtgata aattagctat cgagagatgc caagctgttc ttagtggtgc ggttaatgaa
      901 gtgtctgcgc aaggtttaga ggacagcagt gagttttata aagagataaa atcaagaaag
      961 aacgatatta aaggattttc ggtaagcgat attctaaaga aggactacaa acaattcaat
     1021 ttccaaggaa agggacacaa agggttagag attggtcttt catcaatagt aaaaagaatg
     1081 tcttggctat tcaatgaaca cggtggtgaa gcagatttcg tcaaccaatg cagaagattt
     1141 caggcggaga gggggctcga tgtattggtt ctgttgactt catggaggaa agctggtgat
     1201 tcacacagag aattggtcat attgggagac tctaacgtgg tacgtgaact cattgaaagg
     1261 gttagcgaca agctccaact tcaattattt gggggcaatc ttgatggagg tgtggcgatg
     1321 tttaagcaac tgaacgtcga ggccaccaga aagcaagtcg tcccctattt agaggaagcg
     1381 tactcaaacc tggaagagtg aggatccggc tgctaacaaa gcccgaaagg aagctgagtt
     1441 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1501 gaggggtttt ttgctgaaag gaggaactat atccggatat cccgcaagag gcccggcagt
     1561 accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg
     1621 agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa
     1681 ctaccgcatt aaagcttatc gatgataagc tgtcaaacat gagaattctt gaagacgaaa
     1741 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1801 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1861 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1921 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1981 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2041 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2101 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2161 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     2221 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2281 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2341 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2401 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2461 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2521 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2581 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2641 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2701 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2761 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2821 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2881 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2941 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3001 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3061 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3121 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3181 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3241 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3301 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3361 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3421 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3481 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3541 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3601 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3661 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3721 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3781 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     3841 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     3901 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3961 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     4021 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     4081 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     4141 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     4201 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     4261 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     4321 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     4381 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     4441 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     4501 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     4561 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     4621 cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc
     4681 tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc
     4741 tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc
     4801 ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca
     4861 gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc
     4921 ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc
     4981 gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
     5041 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
     5101 gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat
     5161 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     5221 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     5281 gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     5341 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     5401 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     5461 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5521 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     5581 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     5641 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     5701 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     5761 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     5821 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     5881 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     5941 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     6001 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     6061 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     6121 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     6181 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     6241 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     6301 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     6361 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     6421 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     6481 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     6541 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     6601 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     6661 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     6721 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     6781 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     6841 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc g
//
