LOCUS       sequence-462857-        5879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC18 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5879)
  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 5879)
  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-ZDHHC18
FEATURES             Location/Qualifiers
     source          1..5879
                     /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    2220..2842
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2419..2438)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2866..2886)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2871..3200
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3051..3186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3238..3257)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3239..3257)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3271..3287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3271..3287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3284..3306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3295..3311
                     /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(3319..3325,3326..3343,3344..3349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3364..3385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3502..3519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3673..4261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3753..3772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4432..5223,5224..5292))
                     /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     5055..5074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5293..5397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5424..5852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5511..5530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5721..5742
                     /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 ttcaatgaag
     1021 gactgcgagt accagcagat cagccccggg gccgccccgc tgcccgcctc cccgggggcg
     1081 cgccgtcccg gccccgccgc gtccccgact ccgggccccg ggcccgcgcc gcccgccgcc
     1141 cccgccccgc cgcgctggag cagcagcggc agcggcagcg gcagcgggag cgggagcctc
     1201 ggccgccgcc cacggcgcaa gtgggaggtg ttcccgggtc gcaatcgctt ctactgcggc
     1261 ggccgcctca tgctggccgg ccacggcggc gtcttcgcgc tcacgctgct gctcatcctc
     1321 accaccaccg gcctcttctt cgtctttgac tgtccctacc tggctcgcaa gctgaccctt
     1381 gccatcccca tcatcgctgc catcctcttc ttcttcgtca tgagctgcct gctgcagaca
     1441 agcttcaccg accctgggat cctgccccgg gccactgtct gtgaagcagc cgccctggag
     1501 aaacagatcg acaacacagg cagttctaca taccggccac cccctcggac ccgggaggtg
     1561 ctgatcaacg ggcagatggt gaagctgaag tactgcttca cctgcaagat gttccggcca
     1621 ccccgaacct cacactgcag tgtctgcgac aactgtgtgg aacgatttga ccatcactgc
     1681 ccctgggtgg gcaactgtgt ggggagacgg aactatcgct tcttctacgc gtttattctc
     1741 tccctctcat tcctgacggc cttcatcttc gcctgtgtgg tcacccacct gacgttgcgc
     1801 gctcagggaa gcaacttcct ctccactctg aaggagacac cagcaagcgt gctggagttg
     1861 gtgatctgct tcttctccat ctggtccatt ctgggcctct cagggtttca cacgtacctc
     1921 gtcgcctcca acctgactac taatgaagac atcaaaggct cgtggtccag caagaggggc
     1981 ggtgaggcct ctgtcaaccc ctacagccat aaaagtatta tcaccaactg ctgtgctgtg
     2041 ctctgtggcc ccctacctcc cagcctaatt gaccggaggg gatttgtgca gtccgacacc
     2101 gtgttgccct cacccatcag aagcgatgag ccagcctgca gagccaagcc tgatgccagc
     2161 atggtaggag gccacccctg aagatctgat atcggtacca gtcgactcta gaggatcccg
     2221 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
     2281 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
     2341 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
     2401 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
     2461 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
     2521 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
     2581 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
     2641 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
     2701 ttttaaaata actataccag caggaggacg tccagacaca gcataggcta cctggccatg
     2761 cccaaccggt gggacatttg agttgcttgc ttggcactgt cctctcatgc gttgggtcca
     2821 ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg gctgtggaat gtgtgtcagt
     2881 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2941 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3001 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3061 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3121 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3181 gaggcctagg cttttgcaaa aagctcctcg aggaactgaa aaaccagaaa gttaattccc
     3241 tatagtgagt cgtattaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
     3301 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3361 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3421 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3481 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3541 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3601 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3661 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3721 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3781 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3841 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3901 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3961 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4021 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4081 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4141 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4201 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4261 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4321 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4381 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4501 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4621 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4861 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4921 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5101 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5281 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5341 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5401 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5461 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5521 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5581 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5641 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5701 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5761 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5821 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattt
//
