LOCUS       sequence-462830-        5807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC9 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5807)
  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 5807)
  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-ZDHHC9
FEATURES             Location/Qualifiers
     source          1..5807
                     /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    2148..2770
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2347..2366)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2794..2814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2799..3128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2979..3114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3166..3185)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3167..3185)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3199..3215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3199..3215)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3212..3234)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3223..3239
                     /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(3247..3253,3254..3271,3272..3277))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3292..3313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3430..3447)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3601..4189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3681..3700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4360..5151,5152..5220))
                     /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     4983..5002
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5221..5325)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5352..5780
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5439..5458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5649..5670
                     /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 ttcaatgtct
     1021 gtgatggtgg tgagaaagaa ggtgacacgg aaatgggaga aactcccagg caggaacacc
     1081 ttttgctgtg atggccgcgt catgatggcc cggcaaaagg gcattttcta cctgaccctt
     1141 ttcctcatcc tggggacatg tacactcttc ttcgcctttg agtgccgcta cctggctgtt
     1201 cagctgtctc ctgccatccc tgtatttgct gccatgctct tccttttctc catggctaca
     1261 ctgttgagga ccagcttcag tgaccctgga gtgattcctc gggcgctacc agatgaagca
     1321 gctttcatag aaatggagat agaagctacc aatggtgcgg tgccccaggg ccagcgacca
     1381 ccgcctcgta tcaagaattt ccagataaac aaccagattg tgaaactgaa atactgttac
     1441 acatgcaaga tcttccggcc tccccgggcc tcccattgca gcatctgtga caactgtgtg
     1501 gagcgcttcg accatcactg cccctgggtg gggaattgtg ttggaaagag gaactaccgc
     1561 tacttctacc tcttcatcct ttctctctcc ctcctcacaa tctatgtctt cgccttcaac
     1621 atcgtctatg tggccctcaa atctttgaaa attggcttct tggagacatt gaaagaaact
     1681 cctggaactg ttctagaagt cctcatttgc ttctttacac tctggtccgt cgtgggactg
     1741 actggatttc atactttcct cgtggctctc aaccagacaa ccaatgaaga catcaaagga
     1801 tcatggacag ggaagaatcg cgtccagaat ccctacagcc atggcaatat tgtgaagaac
     1861 tgctgtgaag tgctgtgtgg ccccttgccc cccagtgtgc tggatcgaag gggtattttg
     1921 ccactggagg aaagtggaag tcgacctccc agtactcaag agaccagtag cagcctcttg
     1981 ccacagagcc cagcccccac agaacacctg aactcaaatg agatgccgga ggacagcagc
     2041 actcccgaag agatgccacc tccagagccc ccagagccac cacaggaggc agctgaagct
     2101 gagaagtaga gatctgatat cggtaccagt cgactctaga ggatcccggg tggcatccct
     2161 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     2221 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     2281 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     2341 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     2401 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     2461 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     2521 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     2581 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac
     2641 tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg
     2701 gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc
     2761 ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa
     2821 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2881 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2941 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3001 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3061 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3121 tttgcaaaaa gctcctcgag gaactgaaaa accagaaagt taattcccta tagtgagtcg
     3181 tattaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3241 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3301 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3361 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3421 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3481 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3541 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3601 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3661 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3721 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3781 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3841 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3901 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3961 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4021 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4081 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4141 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4201 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4261 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4321 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4381 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4441 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4501 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4561 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4621 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4681 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4741 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4801 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4861 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4921 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4981 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5041 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5101 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5161 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5221 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5281 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5341 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5401 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5461 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5521 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5581 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5641 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5701 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5761 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaattt
//
