LOCUS       sequence-462851-        5516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC12 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5516)
  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 5516)
  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-ZDHHC12
FEATURES             Location/Qualifiers
     source          1..5516
                     /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"
     polyA_signal    1857..2479
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2056..2075)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2503..2523)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2508..2837
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2688..2823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2875..2894)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2876..2894)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2908..2924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2908..2924)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2921..2943)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2932..2948
                     /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(2956..2962,2963..2980,2981..2986))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3001..3022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3139..3156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3310..3898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3390..3409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4069..4860,4861..4929))
                     /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     4692..4711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4930..5034)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5061..5489
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5148..5167)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5358..5379
                     /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 ttcaatggcg
     1021 ccctgggcgc tcctcagccc tggggtcctg gtgcggaccg ggcacaccgt gctgacctgg
     1081 ggaatcacgc tggtgctctt cctgcacgat accgagctgc ggcaatggga ggagcagggg
     1141 gagctgctcc tgcccctcac cttcctgctc ctggtgctgg gctccctgct gctctacctc
     1201 gctgtgtcac tcatggaccc tggctacgtg aatgtgcagc cccagcctca ggaggagctc
     1261 aaagaggagc agacagccat ggttcctcca gccatccctc ttcggcgctg cagatactgc
     1321 ctggtgctgc agcccctgag ggctcggcac tgccgtgagt gccgccgttg cgtccgccgc
     1381 tacgaccacc actgcccctg gatggagaac tgtgtgggag agcgcaacca cccactcttt
     1441 gtggtctacc tggcgctgca gctggtggtg cttctgtggg gcctgtacct ggcatggtca
     1501 ggcctccggt tcttccagcc ctggggtctg tggttgcggt ccagcgggct cctgttcgcc
     1561 accttcctgc tgctgtccct cttctcgttg gtggccagcc tgctcctcgt ctcgcacctc
     1621 tacctggtgg ccagcaacac caccacctgg gaattcatct cctcacaccg catcgcctat
     1681 ctccgccagc gccccagcaa ccccttcgac cgaggcctga cccgcaacct ggcccacttc
     1741 ttctgtggat ggccctcagg gtcctgggag accctctggg ctgaggagga ggaagagggc
     1801 agcagcccag ctgtttagag atctgatatc ggtaccagtc gactctagag gatcccgggt
     1861 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     1921 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     1981 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     2041 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     2101 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     2161 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     2221 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     2281 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     2341 taaaataact ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc
     2401 aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc
     2461 agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag
     2521 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2581 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2641 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2701 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2761 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2821 gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat
     2881 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     2941 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3001 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3061 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3121 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3181 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3241 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3301 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3361 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3421 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3481 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3541 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3601 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3661 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3721 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3781 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3841 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3901 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3961 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4021 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4081 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4141 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4201 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4261 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4321 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4381 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4441 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4501 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4561 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4621 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4681 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4741 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4801 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4861 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4921 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4981 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5041 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5101 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5161 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5221 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5281 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5341 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5401 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5461 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caattt
//
