LOCUS       sequence-462839-        5726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC15 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5726)
  AUTHORS   Zhang Z, Ren C, Xiao R, Ma S, Liu H, Dou Y, Fan Y, Wang S, Zhan P, 
            Gao C, Yue X, Li C, Gao L, Liang X, Wu Z, Ma C
  TITLE     Palmitoylation of TIM-3 promotes immune exhaustion and restrains 
            antitumor immunity.
  JOURNAL   Sci Immunol. 2024 Nov 15;9(101):eadp7302. doi: 
            10.1126/sciimmunol.adp7302. Epub 2024 Nov 15.
  PUBMED    39546589
REFERENCE   2  (bases 1 to 5726)
  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 
            Immunol."; date: "2024-11-15"; volume: "9(101):eadp7302. doi"; 
            pages: " 10.1126/sciimmunol.adp7302. Epub 2024 Nov 15"
COMMENT     Sequence Label: Flag-ZDHHC15
FEATURES             Location/Qualifiers
     source          1..5726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1532..1543)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2067..2689
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2266..2285)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2713..2733)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2718..3047
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2898..3033
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3085..3104)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3086..3104)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3118..3134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3118..3134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3131..3153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3142..3158
                     /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(3166..3172,3173..3190,3191..3196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3211..3232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3349..3366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3520..4108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3600..3619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4279..5070,5071..5139))
                     /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     4902..4921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5140..5244)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5271..5699
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5358..5377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5568..5589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcaatgcgg
     1021 cgaggctgga agatggctct gtctgggggg ctgcggtgct gccgccgggt actgtcctgg
     1081 gtgccagtgc tcgttattgt cctcgtcgtg ctctggtcct actatgccta cgtctttgaa
     1141 ctctgcctgg tgactgtttt gagcccagca gaaaaagtta tttacctcat actctaccat
     1201 gccatctttg tgttctttac ctggacctac tggaagtcta tctttacact cccacagcag
     1261 ccaaaccaga agttccactt gtcctacaca gacaaggagc gctatgaaaa tgaagaaaga
     1321 cctgaggtcc agaagcagat gcttgttgat atggccaaaa agctaccggt ttacacaaga
     1381 actggaagtg gagctgtacg attctgtgac cggtgtcatc tgatcaagcc agaccgctgc
     1441 caccactgct ctgtctgtgc tatgtgtgtg ttaaaaatgg atcatcactg cccttgggtt
     1501 aataactgca ttggattttc caactacaaa ttcttccttc aattcttagc ttactctgtt
     1561 ctctactgcc tgtacattgc tacgacagtc ttcagctatt tcatcaaata ctggagaggg
     1621 gaattaccca gtgttcgctc taagttccat gtcctttttc ttctctttgt ggcctgcatg
     1681 ttttttgtca gccttgtgat tctctttggt taccattgtt ggcttgtcag cagaaacaaa
     1741 accaccttag aggccttctg cactccagtg tttacaagtg gcccagagaa aaatgggttc
     1801 aaccttggct tcatcaagaa tatccagcag gtgtttggag ataagaagaa gttctggtta
     1861 atacctattg gttccagccc tggtgatgga cactccttcc ctatgaggtc tatgaatgag
     1921 tcacagaacc cactgctagc aaatgaagaa acctgggaag acaacgagga tgacaaccaa
     1981 gattatccag aaggctcatc atctcttgct gtggaaacgg aaacatagag atctgatatc
     2041 ggtaccagtc gactctagag gatcccgggt ggcatccctg tgacccctcc ccagtgcctc
     2101 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     2161 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     2221 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     2281 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     2341 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     2401 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     2461 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     2521 ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc
     2581 agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg
     2641 gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc
     2701 cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2761 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2821 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2881 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2941 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3001 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg
     3061 aactgaaaaa ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc
     3121 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3181 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3241 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3301 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3361 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3421 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3481 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3661 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3961 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4381 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4861 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4921 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4981 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5041 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5101 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     5281 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5341 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5401 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5461 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5521 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5581 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5641 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5701 taacaaaata ttaacgctta caattt
//
