LOCUS       sequence-212702-        6894 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 6894)
  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 6894)
  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..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1522..1539)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1549..1566)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1585..1607
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1622..1646
                     /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     complement(1646..1665)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1647..1665)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1749..1768)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1974..2051
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2052..3134
                     /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(2071..2090)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4446..4637
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4631..4653)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4739..4881
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4794..4813
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4896..4913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5067..5655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5147..5166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5826..6617,6618..6686))
                     /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     6449..6468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6687..6791)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6859..6877
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atcctcactc ttccaggttt gagtacgctt cctctaaata
      361 ggggacgact tgctttctgg tggcctcgac gttcagttgc ttaaacatcg ccacacctcc
      421 atcaagattg cccccaaata attgaagttg gagcttgtcg ctaacccttt caatgagttc
      481 acgtaccacg ttagagtctc ccaatatgac caattctctg tgtgaatcac cagctttcct
      541 ccatgaagtc aacagaacca atacatcgag ccccctctcc gcctgaaatc ttctgcattg
      601 gttgacgaaa tctgcttcac caccgtgttc attgaatagc caagacattc tttttactat
      661 tgatgaaaga ccaatctcta accctttgtg tccctttcct tggaaattga attgtttgta
      721 gtccttcttt agaatatcgc ttaccgaaaa tcctttaata tcgttctttc ttgattttat
      781 ctctttataa aactcactgc tgtcctctaa accttgcgca gacacttcat taaccgcacc
      841 actaagaaca gcttggcatc tctcgatagc taatttatca ctttcctcga ctttgcgcct
      901 catatttgaa gtgtctatta agatggcccc catcaaaagt ggtgcaatgt tcatcaccac
      961 ttcacggtca ccttgcaatt tttcatacca gtagttaaaa accagcgatg agcaactgcc
     1021 ggacactttt acaatccgag gttcagcatc caaatgtttt tgcaagtcaa aatggtggtc
     1081 tataatgcca acgacgttat ctatataatt tttcaaattc tttggtgtat cgttattatc
     1141 taccaagtaa gagtttaatt cagtaccctg ggagacattt tgcttcaagc tctttaaatc
     1201 ttcaatgaag aataattcct cctccttaat tttcagtttt tctaggacat acattacgtc
     1261 tcttcttaaa ctgaggtctt ctctaggaat gtcgataatt gggacaataa agcttccttt
     1321 ctttttctcc tccgagtaag taccttcatt atatatgtat tggcagtacg aataagtgat
     1381 tgcactagca attgagtcca tatctgctga ctcgttacca acacatattg ttaacacatc
     1441 gttgcttgca atttttgaaa ttggcagtga ttttaagtgt gccaaaaatt caggaaccgt
     1501 ctttctcaaa ggcgacatat ggctgccgcg cggcaccagg ccgctgctgt gatgatgatg
     1561 atgatggctg ctgcccatgg tatatctcct tcttaaagtt aaacaaaatt atttctagag
     1621 gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga
     1681 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     1741 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     1801 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     1861 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     1921 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
     1981 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     2041 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     2101 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     2161 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     2221 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     2281 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     2341 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     2401 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     2461 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     2521 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     2581 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     2641 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     2701 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     2761 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     2821 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     2881 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     2941 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     3001 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     3061 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3121 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga
     3181 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     3241 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     3301 ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc
     3361 ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt
     3421 cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg
     3481 ctgggctacg tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt
     3541 cttctcgctt ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta
     3601 gatgacgacc atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg
     3661 atcactggac cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg
     3721 ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt
     3781 gcatggagcc gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca
     3841 ccactccaag aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc
     3901 cttggcagaa catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg
     3961 gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc
     4021 taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt
     4081 gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt
     4141 tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga
     4201 tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc
     4261 gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt
     4321 taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca
     4381 tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg
     4441 catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga
     4501 cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg
     4561 aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg
     4621 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     4681 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     4741 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
     4801 agagcagatt gtactgagag tgcaccatat atgcggtgtg aaataccgca cagatgcgta
     4861 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4921 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4981 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5041 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5101 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5161 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5221 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5281 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5341 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5401 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5461 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5521 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5581 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5641 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5701 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5761 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5821 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5881 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5941 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6001 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6061 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6121 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6181 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6241 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6301 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6361 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6421 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     6481 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6541 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6601 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6661 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6721 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6781 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6841 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaa
//
