LOCUS       sequence-254542-        3150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PGK promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3150)
  AUTHORS   Hazelbaker DZ, Beccard A, Angelini G, Mazzucato P, Messana A, Lam D,
            Eggan K, Barrett LE
  TITLE     A multiplexed gRNA piggyBac transposon system facilitates efficient 
            induction of CRISPRi and CRISPRa in human pluripotent stem cells.
  JOURNAL   Sci Rep. 2020 Jan 20;10(1):635. doi: 10.1038/s41598-020-57500-1.
  PUBMED    31959800
REFERENCE   2  (bases 1 to 3150)
  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: "Sci Rep."; 
            date: "2020-01-20"; volume: "10(1):635. doi"; pages: " 
            10.1038/s41598-020-57500-1"
COMMENT     Sequence Label: S12_EcoRV_PGK_AscI
FEATURES             Location/Qualifiers
     source          1..3150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..56)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     156..175
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     369..391
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     383..400
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     384..400
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(434..933)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(482..501)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     875..898
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(979..995)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(979..995)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(992..1014)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1003..1019
                     /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(1027..1033,1034..1051,1052..1057))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1072..1093
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1210..1227)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1381..1969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1461..1480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2140..2955)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2140..2955)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2863..2882
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2956..3060)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3128..3146
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
       61 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
      121 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
      181 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
      241 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
      301 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      361 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgacgc gtattgggat
      421 gttccggcgc gcccgaaagg cccggagatg aggaagagga gaacagcgcg gcagacgtgc
      481 gcttttgaag cgtgcagaat gccgggcctc gggaggacct tcgggcgccc gccccgcccc
      541 tgagcccgcc cctgagcccg cccccggacc caccccttcc cagcctctga gcccagaaag
      601 cgaaggagca aagctgctat tggccgctgc cccaaaggcc tacccgcttc cattgctcag
      661 cggtgctgtc catctgcacg agactagtga gacgtgctac ttccatttgt cacgtcctgc
      721 acgacgcgag ctgcggggcg ggggggaact tcctgactag gggaggagta gaaggtggcg
      781 cgaaggggcc accaaagaac ggagccggtt ggcgcctacc ggtggatgtg gaatgtgtgc
      841 gaggccagag gccacttgtg tagcgccaag tgcccagcgg ggctgctaaa gcgcatgctc
      901 cagactgcct tgggaaaagc gcctccccta cccgatatcg ttccatccca atggcgcgcc
      961 gagcttggct cgagcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1021 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1081 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1141 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1201 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1261 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1321 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1381 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1441 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1501 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1561 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1621 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1681 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1741 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1801 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1861 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1921 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1981 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2041 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2101 atcaatctaa agtatatatg agtaaacttg gtctgacagt tagaaaaact catcgagcat
     2161 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     2221 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     2281 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     2341 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     2401 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     2461 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     2521 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     2581 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     2641 tgttttccca gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     2701 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     2761 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     2821 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     2881 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg
     2941 ttgaatatgg ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3001 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3061 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     3121 ctataaaaat aggcgtatca cgaggccctt
//
