LOCUS       sequence-462840-        5612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC3 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5612)
  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 5612)
  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-ZDHHC3
FEATURES             Location/Qualifiers
     source          1..5612
                     /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    1953..2575
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2152..2171)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2599..2619)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2604..2933
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2784..2919
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2971..2990)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2972..2990)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3004..3020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3004..3020)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3017..3039)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3028..3044
                     /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(3052..3058,3059..3076,3077..3082))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3097..3118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3235..3252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3406..3994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3486..3505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4165..4956,4957..5025))
                     /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     4788..4807
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5026..5130)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5157..5585
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5244..5263)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5454..5475
                     /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 ttcaatgatg
     1021 cttatcccca cccaccactt ccgaaacatt gagcggaaac cagaatacct ccagccagag
     1081 aagtgtgtcc caccccccta ccctggtcct gtgggaacca tgtggtttat ccgtgacggc
     1141 tgtggcatcg cctgtgccat cgttacctgg tttctggtcc tctatgcgga gttcgtggtc
     1201 ctctttgtca tgctgattcc atctcgagac tacgtgtata gcatcatcaa cggaattgtg
     1261 ttcaacctgc tggccttctt ggccctggcc tcccactgcc gggccatgct gacggacccc
     1321 ggggcagtgc ccaaaggaaa tgccactaaa gaattcatcg agagtttaca gttgaagcct
     1381 gggcaggtgg tgtacaagtg ccccaaatgc tgcagcatca agcccgaccg agcccaccac
     1441 tgcagtgttt gtaagcggtg cattcggaag atggaccacc actgtccctg ggtcaacaac
     1501 tgtgtaggcg agaacaacca gaagtacttc gtcctgttta caatgtacat agctctcatt
     1561 tccttgcacg ccctcatcat ggtgggattc cacttcctgc attgctttga agaagattgg
     1621 acaaagtgca gctccttctc tccacccacc acagtgattc tccttatcct gctgtgcttt
     1681 gagggcctgc tcttcctcat tttcacatca gtgatgtttg ggacccaggt gcactccatc
     1741 tgcacagatg agacgggaat agaacaattg aaaaaggaag agagaagatg ggctaaaaaa
     1801 acaaaatgga tgaacatgaa agccgttttt ggccacccct tctctctagg ctgggccagc
     1861 ccctttgcca cgccagacca agggaaggca gacccgtacc agtatgtggt ctgaagatct
     1921 gatatcggta ccagtcgact ctagaggatc ccgggtggca tccctgtgac ccctccccag
     1981 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     2041 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     2101 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     2161 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     2221 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     2281 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     2341 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     2401 cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg
     2461 acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct
     2521 tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta
     2581 cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2641 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2701 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2761 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2821 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     2881 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     2941 tcgaggaact gaaaaaccag aaagttaatt ccctatagtg agtcgtatta aattcgtaat
     3001 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3061 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3121 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3181 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3241 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3301 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3361 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3421 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3481 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3541 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3601 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3661 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3721 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3781 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3841 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3901 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3961 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4021 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4081 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4141 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4201 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4261 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4321 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4381 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4441 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4501 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4561 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4621 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4681 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4741 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4801 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4861 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4921 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4981 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5041 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5101 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     5161 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5221 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5281 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5341 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5401 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5461 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5521 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5581 gaattttaac aaaatattaa cgcttacaat tt
//
