LOCUS       sequence-462864-        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC20 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  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 5810)
  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-ZDHHC20
FEATURES             Location/Qualifiers
     source          1..5810
                     /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    2151..2773
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2350..2369)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2797..2817)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2802..3131
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2982..3117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3169..3188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3170..3188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3202..3218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3202..3218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3215..3237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3226..3242
                     /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(3250..3256,3257..3274,3275..3280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3295..3316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3433..3450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3604..4192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3684..3703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4363..5154,5155..5223))
                     /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     4986..5005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5224..5328)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5355..5783
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5442..5461)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5652..5673
                     /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 ccctggacgc tgtggcgctg ctgccagcgc gtcgtgggct gggtgccggt gctcttcatc
     1081 accttcgtgg tcgtctggtc ctactacgcg tacgtggtgg agctctgcgt gtttactatt
     1141 tttggaaatg aagaaaatgg aaagaccgtt gtttaccttg tggctttcca tctgttcttt
     1201 gttatgtttg tatggtccta ttggatgaca attttcacat ctcccgcttc cccctccaaa
     1261 gagttctact tgtccaattc tgaaaaggaa cgttatgaaa aagaattcag ccaagaaaga
     1321 caacaagaaa ttttgagaag agcagcaaga gctttaccta tctataccac atcagcttca
     1381 aaaactatca gatattgtga aaaatgtcag ctgattaaac ctgatcgggc gcatcactgc
     1441 tcagcctgtg actcatgtat tcttaagatg gatcatcact gtccttgggt gaataactgt
     1501 gtgggatttt ctaattacaa attcttcctg ctgtttttat tgtattccct attatattgc
     1561 cttttcgtgg ctgcaacagt tttagagtac tttataaaat tttggacgaa tgaactgaca
     1621 gatacacgtg caaaattcca cgtacttttt cttttctttg tgtctgcaat gttcttcatc
     1681 agcgtcctct cacttttcag ctaccactgc tggctagttg gaaaaaatag aacaacaata
     1741 gaatcattcc gcgcacccac gttttcatac ggacctgatg gaaatggttt ctctcttgga
     1801 tgcagtaaaa attggagaca agtctttggt gatgaaaaga aatattggct acttccaata
     1861 ttttcaagct tgggtgatgg ttgcagtttt ccaactcgcc ttgtggggat ggatccagaa
     1921 caagcttctg ttacaaacca gaatgagtat gccagaagta gtggctcaaa tcaacctttt
     1981 cctatcaaac cacttagtga atcaaaaaac cgcttgttgg acagtgaatc tcagtggctg
     2041 gagaatggag ctgaagaagg catcgtcaaa tcagggacaa ataaccatgt aacagtggca
     2101 atagaaaatt gaagatctga tatcggtacc agtcgactct agaggatccc gggtggcatc
     2161 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     2221 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     2281 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     2341 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     2401 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     2461 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     2521 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     2581 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
     2641 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
     2701 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
     2761 tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg
     2821 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2881 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2941 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3001 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3061 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3121 gcttttgcaa aaagctcctc gaggaactga aaaaccagaa agttaattcc ctatagtgag
     3181 tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3241 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3301 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3361 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3421 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4021 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4261 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4321 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4441 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4501 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4561 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4801 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4861 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5041 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5221 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5281 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5341 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5401 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5461 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5521 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5581 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5641 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5701 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5761 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
//
