LOCUS       sequence-462843-        5747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC4 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5747)
  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 5747)
  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-ZDHHC4
FEATURES             Location/Qualifiers
     source          1..5747
                     /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    2088..2710
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2287..2306)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2734..2754)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2739..3068
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2919..3054
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3106..3125)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3107..3125)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3139..3155)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3139..3155)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3152..3174)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3163..3179
                     /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(3187..3193,3194..3211,3212..3217))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3232..3253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3370..3387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3541..4129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3621..3640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4300..5091,5092..5160))
                     /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     4923..4942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5161..5265)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5292..5720
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5379..5398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5589..5610
                     /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 ttcaatggac
     1021 tttctggtcc tcttcttgtt ctacctggct tcggtgctga tgggtcttgt tcttatctgc
     1081 gtctgctcga aaacccatag cttgaaaggc ctggccaggg gaggagcaca gatattttcc
     1141 tgtataattc cagaatgtct tcagagagcc gtgcatggat tgcttcatta ccttttccat
     1201 acgagaaacc acaccttcat tgtcctgcac ctggtcttgc aagggatggt ttatactgag
     1261 tacacctggg aagtatttgg ctactgtcag gagctggagt tgtccttgca ttaccttctt
     1321 ctgccctatc tgctgctagg tgtaaacctg ttttttttca ccctgacttg tggaaccaat
     1381 cctggcatta taacaaaagc aaatgaatta ttatttcttc atgtttatga atttgatgaa
     1441 gtgatgtttc caaagaacgt gaggtgctct acttgtgatt taaggaaacc agctcgatcc
     1501 aagcactgca gtgtgtgtaa ctggtgtgtg caccgtttcg accatcactg tgtttgggtg
     1561 aacaactgca tcggggcctg gaacatcagg tacttcctca tctacgtctt gaccttgacg
     1621 gcctcggctg ccaccgtcgc cattgtgagc accacttttc tggtccactt ggtggtgatg
     1681 tcagatttat accaggagac ttacatcgat gaccttggac acctccatgt tatggacacg
     1741 gtctttctta ttcagtacct gttcctgact tttccacgga ttgtcttcat gctgggcttt
     1801 gtcgtggttc tgagcttcct cctgggtggc tacctgttgt ttgtcctgta tctggcggcc
     1861 accaaccaga ctactaacga gtggtacaga ggtgactggg cctggtgcca gcgttgtccc
     1921 cttgtggcct ggcctccgtc agcagagccc caagtccacc ggaacattca ctcccatggg
     1981 cttcggagca accttcaaga gatctttcta cctgcctttc catgtcatga gaggaagaaa
     2041 caagaatgaa gatctgatat cggtaccagt cgactctaga ggatcccggg tggcatccct
     2101 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     2161 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     2221 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     2281 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     2341 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     2401 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     2461 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     2521 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac
     2581 tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg
     2641 gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc
     2701 ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa
     2761 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2821 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2881 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2941 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3001 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3061 tttgcaaaaa gctcctcgag gaactgaaaa accagaaagt taattcccta tagtgagtcg
     3121 tattaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3181 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3241 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3301 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3361 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3421 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3481 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3541 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3601 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3661 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3721 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3781 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3841 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3901 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3961 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4021 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4081 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4141 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4201 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4261 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4321 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4381 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4441 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4501 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4561 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4621 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4681 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4741 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4801 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4861 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4921 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4981 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5041 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5101 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5161 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5221 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5281 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5341 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5401 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5461 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5521 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5581 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5641 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5701 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaattt
//
