LOCUS       sequence-311568-        7094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a HA-tagged E. coli PPK1 in mammalian cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7094)
  AUTHORS   Bondy-Chorney E, Abramchuk I, Nasser R, Holinier C, Denoncourt A, 
            Baijal K, McCarthy L, Khacho M, Lavallee-Adam M, Downey M
  TITLE     A Broad Response to Intracellular Long-Chain Polyphosphate in Human 
            Cells.
  JOURNAL   Cell Rep. 2020 Oct 27;33(4):108318. doi: 
            10.1016/j.celrep.2020.108318.
  PUBMED    33113373
REFERENCE   2  (bases 1 to 7094)
  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: "Cell Rep.";
            date: "2020-10-27"; volume: "33(4):108318. doi"; pages: " 
            10.1016/j.celrep.2020.108318"
COMMENT     Sequence Label: HA-EcPPK1
FEATURES             Location/Qualifiers
     source          1..7094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             981..1007
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3102..3119)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3108..3332
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3378..3806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3465..3484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3675..3696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3815..3835)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3820..4149
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4000..4135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4062..4081
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4197..4244
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4263..4637
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4767..4888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4804..4823)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4858..4877
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4937..4953)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4937..4953)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4950..4972)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4961..4977
                     /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)."
     promoter        complement(join(4985..4991,4992..5009,5010..5015))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5030..5051
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5168..5185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5339..5927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5419..5438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6098..6889,6890..6958))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6721..6740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6959..7063)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc
      961 accacactgg actagtgatg tacccatacg atgttccaga ttacgctggg gggatgggtc
     1021 aggaaaagct atacatcgaa aaagagctca gttggttatc gttcaatgaa cgcgtgcttc
     1081 aggaagcggc ggacaaatct aacccgctga ttgaaaggat gcgtttcctg gggatctatt
     1141 ccaataacct tgatgagttc tataaagtcc gcttcgctga actgaagcga cgcatcatta
     1201 ttagcgaaga acaaggctcc aactctcatt cccgccattt actgggcaaa attcagtccc
     1261 gggtgctgaa agccgatcag gaattcgacg gcctctacaa cgagctattg ctggagatgg
     1321 cgcgcaacca gatcttcctg attaatgaac gccagctctc cgtcaatcaa caaaactggc
     1381 tgcgtcatta ttttaagcag tatctgcgtc agcacattac gccgatttta atcaatcctg
     1441 acactgactt agtgcagttc ctgaaagatg attacaccta tctggcggtg gaaattatcc
     1501 gtggcgatac catccgttac gcgctgctgg agatcccatc agataaagtg ccgcgctttg
     1561 tgaatttacc gccagaagcg ccgcgtcgac gcaagccgat gattcttctg gataacattc
     1621 tgcgttactg ccttgatgat attttcaaag gcttctttga ttatgacgcg ctgaatgcct
     1681 attcaatgaa gatgacccgc gatgccgaat acgatttagt gcatgagatg gaagccagcc
     1741 tgatggagtt gatgtcttcc agtctcaagc agcgtttaac tgctgagccg gtgcgttttg
     1801 tttatcagcg cgatatgccc aatgcgctgg ttgaagtgtt acgcgaaaaa ctgactattt
     1861 cccgctacga ctccatcgtc cccggcggtc gttatcataa ttttaaagac tttattaatt
     1921 tccccaatgt cggcaaagcc aatctggtga acaaaccact gccgcgttta cgccatattt
     1981 ggtttgataa agcccagttc cgcaatggtt ttgatgccat tcgcgaacgc gatgtgttgc
     2041 tctattatcc ttatcacacc tttgagcatg tgctggaact gctgcgtcag gcttcgttcg
     2101 acccgagcgt actggcgatt aaaattaaca tttaccgcgt ggcgaaagat tcacgcatca
     2161 tcgactcgat gatccacgcc gcacataacg gtaagaaagt caccgtggtg gttgagttac
     2221 aggcgcgttt cgacgaagaa gccaacattc actgggcgaa gcgcctgacc gaagcaggcg
     2281 tgcacgttat cttctctgcg ccggggctga aaattcacgc caaactgttc ctgatttcac
     2341 gtaaagaaaa cggtgaagtg gtgcgttatg cacacatcgg gaccgggaac tttaacgaaa
     2401 aaaccgcgcg tctttatact gactattcgt tgctgaccgc cgatgcgcgc atcaccaacg
     2461 aagtacggcg ggtatttaac tttattgaaa acccataccg tccggtgaca tttgattatt
     2521 taatggtatc gccgcaaaac tcccgccgcc tattgtatga aatggtggac cgcgagatcg
     2581 ccaacgcgca gcaagggctg cccagtggta tcaccctgaa gctaaataac cttgtcgata
     2641 aaggcctggt tgatcgtctg tatgcggcct ccagctccgg cgtaccggtt aatctgctgg
     2701 ttcgcggaat gtgttcgctg atccccaatc tggaaggcat tagcgacaac attcgtgcca
     2761 tcagtattgt tgaccgttac cttgaacatg accgggttta tatttttgaa aatggcggcg
     2821 ataaaaaggt ctacctttct tccgccgact ggatgacgcg caatattgat tatcgtattg
     2881 aagtggcgac gccgctgctc gatccgcgcc tgaagcagcg ggtactggac atcattgaca
     2941 tattgttcag cgatacggtc aaagcacgtt atatcgataa agaactcagt aatcgctacg
     3001 ttccccgcgg caatcgccgc aaagtacggg cgcagttggc gatttatgac tacatcaaat
     3061 cactcgaaca acctgaataa taagtttaaa ccgctgatca gcctcgactg tgccttctag
     3121 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3181 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3241 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3301 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3361 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3421 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3721 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3781 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3841 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3901 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3961 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4021 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4081 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4141 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
     4201 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     4261 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
     4321 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
     4381 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
     4441 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
     4501 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
     4561 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
     4621 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     4681 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4741 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4801 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4861 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4921 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4981 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5041 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5101 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5161 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5221 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5281 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5341 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5401 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5461 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5521 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5581 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5641 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5701 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5761 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5821 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5881 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5941 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6001 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6061 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6181 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6241 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6301 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6481 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6541 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6601 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6781 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6961 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7021 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7081 tgccacctga cgtc
//
