LOCUS       sequence-465267-        5807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC9-C169S 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-C169S
FEATURES             Location/Qualifiers
     source          1..5807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(77..406)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      91..226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     391..411
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    435..1057
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     839..858
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2209..2274)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(2304..2507)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2333..2353)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2508..2887
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3048..3065)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3048..3064)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3057..3079)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3205..3660)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3342..3363)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3554..3573
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3687..3791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3792..3860,3861..4652)
                     /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     complement(4010..4029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5312..5331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5565..5582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5699..5720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5735..5740,5741..5758,5759..5765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5773..5789
                     /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)."
     primer_bind     5778..5800
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5797..5807,1..6)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5797..5807,1..6)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tatgacatga ttacgaattt aatacgactc actataggga attaactttc tggtttttca
       61 gttcctcgag gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct
      121 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
      181 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
      241 gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
      301 agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc
      361 tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc cacagccaag
      421 ctggccgcgt acccaattca acaggcatct actgagtgga cccaacgcat gagaggacag
      481 tgccaagcaa gcaactcaaa tgtcccaccg gttgggcatg gccaggtagc ctatgctgtg
      541 tctggacgtc ctcctgctgg tatagttatt ttaaaatcag aaggacaggg aagggagcag
      601 tggttcacgc ctgtaatccc agcaatttgg gaggccaagg tgggtagatc acctgagatt
      661 aggagttgga gaccagcctg gccaatatgg tgaaaccccg tctctaccaa aaaaacaaaa
      721 attagctgag cctggtcatg catgcctgga atcccaacaa ctcgggaggc tgaggcagga
      781 gaatcgcttg aacccaggag gcggagattg cagtgagcca agattgtgcc actgcactcc
      841 agcttggttc ccaatagacc ccgcaggccc tacaggttgt cttcccaact tgccccttgc
      901 tccataccac ccccctccac cccataatat tatagaagga cacctagtca gacaaaatga
      961 tgcaacttaa ttttattagg acaaggctgg tgggcactgg agtggcaact tccagggcca
     1021 ggagaggcac tggggagggg tcacagggat gccacccggg atcctctaga gtcgactggt
     1081 accgatatca gatctctact tctcagcttc agctgcctcc tgtggtggct ctgggggctc
     1141 tggaggtggc atctcttcgg gagtgctgct gtcctccggc atctcatttg agttcaggtg
     1201 ttctgtgggg gctgggctct gtggcaagag gctgctactg gtctcttgag tactgggagg
     1261 tcgacttcca ctttcctcca gtggcaaaat accccttcga tccagcacac tggggggcaa
     1321 ggggccacac agcacttcac agcagttctt cacaatattg ccatggctgt agggattctg
     1381 gacgcgattc ttccctgtcc atgatccttt gatgtcttca ttggttgtct ggttgagagc
     1441 cacgaggaaa gtatgaaatc cagtcagtcc cacgacggac cagagtgtaa agaagcaaat
     1501 gaggacttct agaacagttc caggagtttc tttcaatgtc tccaagaagc caattttcaa
     1561 agatttgagg gccacataga cgatgttgaa ggcgaagaca tagattgtga ggagggagag
     1621 agaaaggatg aagaggtaga agtagcggta gttcctcttt ccaacacaat tccccaccca
     1681 ggggctgtga tggtcgaagc gctccacaca gttgtcacag atgctgcaat gggaggcccg
     1741 gggaggccgg aagatcttgc atgtgtaaca gtatttcagt ttcacaatct ggttgtttat
     1801 ctggaaattc ttgatacgag gcggtggtcg ctggccctgg ggcaccgcac cattggtagc
     1861 ttctatctcc atttctatga aagctgcttc atctggtagc gcccgaggaa tcactccagg
     1921 gtcactgaag ctggtcctca acagtgtagc catggagaaa aggaagagca tggcagcaaa
     1981 tacagggatg gcaggagaca gctgaacagc caggtagcgg cactcaaagg cgaagaagag
     2041 tgtacatgtc cccaggatga ggaaaagggt caggtagaaa atgccctttt gccgggccat
     2101 catgacgcgg ccatcacagc aaaaggtgtt cctgcctggg agtttctccc atttccgtgt
     2161 caccttcttt ctcaccacca tcacagacat tgaattcgcg gccgcaagct tgtcatcgtc
     2221 atccttgtaa tcgatgtcat gatctttata atcaccgtca tggtctttgt agtccatggt
     2281 taattctgac ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta
     2341 ccgcccattt gcgtcaacgg ggcggggtta ttacgacatt ttggaaagtc ccgttgattt
     2401 tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag
     2461 tcaaaccgct atccacgccc attggtgtac tgccaaaacc gcatcaccat ggtaatagcg
     2521 atgactaata cgtagatgta ctgccaagta ggaaagtccc gtaaggtcat gtactgggca
     2581 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcggact tggcatatga
     2641 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     2701 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     2761 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     2821 atatatgggc tatgaactaa tgaccccgta attgattact attaataact agtcaataat
     2881 caatgtcaac atggcggtca tattggacat gagccaatat aaatgtacat attatgatat
     2941 agatacaacg tatgcaatgg ccaatagcca atattgattt atgctatata accaatgact
     3001 aatatggcta attgccaata ttgattcaat gtatagatca gcttggcact ggccgtcgtt
     3061 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3121 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3181 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
     3241 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3301 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3361 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3421 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3481 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3541 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3601 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3661 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4741 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5581 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5641 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5701 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5761 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagc
//
