LOCUS       sequence-209213-        7058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses E. coli PPK1 in mammalian cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7058)
  AUTHORS   Bentley-DeSousa A, Holinier C, Moteshareie H, Tseng YC, Kajjo S, 
            Nwosu C, Amodeo GF, Bondy-Chorney E, Sai Y, Rudner A, Golshani A, 
            Davey NE, Downey M
  TITLE     A Screen for Candidate Targets of Lysine Polyphosphorylation 
            Uncovers a Conserved Network Implicated in Ribosome Biogenesis.
  JOURNAL   Cell Rep. 2018 Mar 27;22(13):3427-3439. doi: 
            10.1016/j.celrep.2018.02.104.
  PUBMED    29590613
REFERENCE   2  (bases 1 to 7058)
  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.celrep.2018.02"; journalName: "Cell Rep"; date: 
            "2018-03-27- 27"; volume: "22"; issue: "13"; pages: "3427-3439"
COMMENT     Sequence Label: EcPPK1
FEATURES             Location/Qualifiers
     source          1..7058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2832..2849)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2838..3062
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3108..3536
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3195..3214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3405..3426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3545..3565)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3550..3879
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3730..3865
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3792..3811
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3927..3974
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3993..4367
                     /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    4497..4618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4534..4553)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4588..4607
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4667..4683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4667..4683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4680..4702)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4691..4707
                     /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(4715..4721,4722..4739,4740..4745))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4760..4781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4898..4915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5069..5657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5149..5168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5828..6619,6620..6688))
                     /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     6451..6470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6689..6793)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6868..6887)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagactcg agcggccgcc actgtgctgg atatctgcag
      721 aattccacca cactggacta gtgatgggtc aggaaaagct atacatcgaa aaagagctca
      781 gttggttatc gttcaatgaa cgcgtgcttc aggaagcggc ggacaaatct aacccgctga
      841 ttgaaaggat gcgtttcctg gggatctatt ccaataacct tgatgagttc tataaagtcc
      901 gcttcgctga actgaagcga cgcatcatta ttagcgaaga acaaggctcc aactctcatt
      961 cccgccattt actgggcaaa attcagtccc gggtgctgaa agccgatcag gaattcgacg
     1021 gcctctacaa cgagctattg ctggagatgg cgcgcaacca gatcttcctg attaatgaac
     1081 gccagctctc cgtcaatcaa caaaactggc tgcgtcatta ttttaagcag tatctgcgtc
     1141 agcacattac gccgatttta atcaatcctg acactgactt agtgcagttc ctgaaagatg
     1201 attacaccta tctggcggtg gaaattatcc gtggcgatac catccgttac gcgctgctgg
     1261 agatcccatc agataaagtg ccgcgctttg tgaatttacc gccagaagcg ccgcgtcgac
     1321 gcaagccgat gattcttctg gataacattc tgcgttactg ccttgatgat attttcaaag
     1381 gcttctttga ttatgacgcg ctgaatgcct attcaatgaa gatgacccgc gatgccgaat
     1441 acgatttagt gcatgagatg gaagccagcc tgatggagtt gatgtcttcc agtctcaagc
     1501 agcgtttaac tgctgagccg gtgcgttttg tttatcagcg cgatatgccc aatgcgctgg
     1561 ttgaagtgtt acgcgaaaaa ctgactattt cccgctacga ctccatcgtc cccggcggtc
     1621 gttatcataa ttttaaagac tttattaatt tccccaatgt cggcaaagcc aatctggtga
     1681 acaaaccact gccgcgttta cgccatattt ggtttgataa agcccagttc cgcaatggtt
     1741 ttgatgccat tcgcgaacgc gatgtgttgc tctattatcc ttatcacacc tttgagcatg
     1801 tgctggaact gctgcgtcag gcttcgttcg acccgagcgt actggcgatt aaaattaaca
     1861 tttaccgcgt ggcgaaagat tcacgcatca tcgactcgat gatccacgcc gcacataacg
     1921 gtaagaaagt caccgtggtg gttgagttac aggcgcgttt cgacgaagaa gccaacattc
     1981 actgggcgaa gcgcctgacc gaagcaggcg tgcacgttat cttctctgcg ccggggctga
     2041 aaattcacgc caaactgttc ctgatttcac gtaaagaaaa cggtgaagtg gtgcgttatg
     2101 cacacatcgg gaccgggaac tttaacgaaa aaaccgcgcg tctttatact gactattcgt
     2161 tgctgaccgc cgatgcgcgc atcaccaacg aagtacggcg ggtatttaac tttattgaaa
     2221 acccataccg tccggtgaca tttgattatt taatggtatc gccgcaaaac tcccgccgcc
     2281 tattgtatga aatggtggac cgcgagatcg ccaacgcgca gcaagggctg cccagtggta
     2341 tcaccctgaa gctaaataac cttgtcgata aaggcctggt tgatcgtctg tatgcggcct
     2401 ccagctccgg cgtaccggtt aatctgctgg ttcgcggaat gtgttcgctg atccccaatc
     2461 tggaaggcat tagcgacaac attcgtgcca tcagtattgt tgaccgttac cttgaacatg
     2521 accgggttta tatttttgaa aatggcggcg ataaaaaggt ctacctttct tccgccgact
     2581 ggatgacgcg caatattgat tatcgtattg aagtggcgac gccgctgctc gatccgcgcc
     2641 tgaagcagcg ggtactggac atcatcgaca tattgttcag cgatacggtc aaagcacgtt
     2701 atatcgataa agaactcagt aatcgctacg ttccccgcgg caatcgccgc aaagtacggg
     2761 cgcagttggc gatttatgac tacatcaaat cactcgaaca acctgaataa taagtttaaa
     2821 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2881 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2941 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3001 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3061 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3121 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3181 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3241 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3301 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3361 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3421 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3481 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3541 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3601 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3661 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3721 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3781 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3841 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3901 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     3961 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     4021 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     4081 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     4141 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     4201 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     4261 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     4321 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     4381 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4441 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4501 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4561 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4621 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4681 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4741 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4801 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4861 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4921 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4981 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5041 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5101 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5161 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5221 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5281 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5341 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5401 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5461 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5521 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5581 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5641 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5701 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5761 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5821 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5881 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5941 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6001 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6061 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6121 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6181 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6241 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6301 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6361 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6421 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6481 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6781 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6841 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6901 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6961 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     7021 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
