LOCUS       sequence-445749-        3230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To downregulate the expression of human DGK alfa in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3230)
  AUTHORS   Avila-Flores A, Arranz-Nicolas J, Andrada E, Soutar D, Merida I
  TITLE     Predominant contribution of DGKzeta over DGKalpha in the control of 
            PKC/PDK-1-regulated functions in T cells.
  JOURNAL   Immunol Cell Biol. 2017 Jul;95(6):549-563. doi: 10.1038/icb.2017.7. 
            Epub 2017 Feb 6.
  PUBMED    28163304
REFERENCE   2  (bases 1 to 3230)
  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: 
            "10.1038/icb.2017.7"; journalName: "Immunol Cell Biol"; date: 
            "2017-07"; volume: "95"; issue: "6"; pages: "549-563"
COMMENT     Sequence Label: pSuper DGK alfa human
FEATURES             Location/Qualifiers
     source          1..3230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     674..690
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     674..690
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        710..924
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     845..864
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(997..1013)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(998..1014)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1042..1060)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1081..1097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1081..1097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1094..1116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1105..1121
                     /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(1129..1135,1136..1153,1154..1159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1174..1195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1312..1329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1483..2071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1563..1582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2242..3033,3034..3102))
                     /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     2865..2884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3103..3207)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg aacgctgacg
      721 tcatcaaccc gctccaagga atcgcgggcc cagtgtcact aggcgggaac acccagcgcg
      781 cgtgcgccct ggcaggaaga tggctgtgag ggacagggga gtggcgccct gcaatatttg
      841 catgtcgcta tgtgttctgg gaaatcacca taaacgtgaa atgtctttgg atttgggaat
      901 cttataagtt ctgtatgaga ccacagatcc ccgccagaag accatggatg attcaagaga
      961 tcatccatgg tcttctggct ttttggaaaa gcttatcgat accgtcgacc tcgagggggg
     1021 gcccggtacc agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg
     1081 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1141 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1201 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1261 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1321 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1381 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1561 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1921 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2221 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2281 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2341 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2401 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2461 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2521 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2581 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2641 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2701 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2761 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2821 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2881 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2941 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3001 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3061 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3121 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3181 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
