LOCUS       sequence-254543-        3932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAMKII1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3932)
  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 3932)
  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: S11_EcoRV_CAMKIIA_AscI
FEATURES             Location/Qualifiers
     source          1..3932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..61)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     161..180
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     374..396
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     388..405
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     389..405
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        432..1721
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     primer_bind     complement(1766..1782)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1766..1782)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1779..1801)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1790..1806
                     /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(1814..1820,1821..1838,1839..1844))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1859..1880
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1997..2014)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2168..2756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2248..2267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2927..3742)
                     /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(2927..3742)
                     /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     3650..3669
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3743..3847)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3915..3932,1)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
       61 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      121 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
      181 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
      241 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      301 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
      361 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt
      421 gggatgatat ccattatggc cttaggtcac ttcatctcca tggggttctt cttctgattt
      481 tctagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag ggtcacatca
      541 gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat gccttggggg
      601 catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca atgagtttag
      661 gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg gtgagtccag
      721 ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga agcacaagca
      781 gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg aggatgggtg
      841 gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga cagatagggg
      901 tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac ttcttctcac
      961 gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg gaaagaaaca
     1021 aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa agaaaccatt
     1081 acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt aaaccttaga
     1141 acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga tgaagttgcc
     1201 agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc cactagccgt
     1261 taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa aagggcccta
     1321 tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag tttgttcaca
     1381 tccccttctc caaccccctc agtacatcac cctgggagaa caaggtccac ttgcttctgg
     1441 gcccacacag tcctgcagta ttgtgtatat aaggccaggg caacggagga gcaggttttg
     1501 aagtgaaagg caggcaggtg ttggggaggc agttaccggg gcaacgggaa cagggcgttt
     1561 cggaggtggt tgccatgggg acctggatgc tgacgaaggc tcgcgaggct gtgagcagcc
     1621 acagtgccct gctcagaagc cccaagctcg tcaatcaagc tggttctcca tttgcactca
     1681 ggagcacggg caggcgagtg gcccctagtt ctgggggcag cggggcgcgc catcccaatg
     1741 gcgcgccgag cttggctcga gcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     1801 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     1861 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     1921 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     1981 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2041 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2101 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2161 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2221 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2281 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2341 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2401 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2461 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2521 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2581 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2641 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2701 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2761 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2821 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2881 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttag aaaaactcat
     2941 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     3001 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     3061 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     3121 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     3181 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     3241 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     3301 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     3361 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     3421 ggaatgctgt tttcccaggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     3481 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     3541 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     3601 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     3661 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     3721 tttcccgttg aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg
     3781 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3841 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     3901 cattaaccta taaaaatagg cgtatcacga gg
//
