LOCUS       sequence-465261-        6860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC5 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6860)
  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 6860)
  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-ZDHHC5
FEATURES             Location/Qualifiers
     source          1..6860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2014..2079)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(2109..2312)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2138..2158)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2313..2692
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2853..2870)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2853..2869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2862..2884)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3010..3465)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3147..3168)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3359..3378
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3492..3596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3597..3665,3666..4457)
                     /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(3815..3834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4628..5216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5117..5136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5370..5387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5504..5525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5540..5545,5546..5563,5564..5570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5578..5594
                     /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     5583..5605
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5602..5618
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5602..5618
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5632..5651
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5632..5650
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5689..6018)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5703..5838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6003..6023
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    6047..6669
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     6451..6470
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
ORIGIN
        1 ctgtacttca tctttgggtg tcagtaatgg ctgcaaggcc acttcttctg tctcagagac
       61 accaggttgg gctttttggc tgctgtaaga catcgatcgg cccaggtatg gggctgttgg
      121 gcctggttca ggggacccta ctccccggcc ccttagccca tctggcttgc caaaacgggg
      181 gacagctggg cgtcccagtg gagtcttgcc caaaggtgat ctctttgaat catctgagga
      241 ggaactagta cgaggggcat ggcccgagcc tggtgttgac tgaatgcctg agtcccccaa
      301 gcctggttct tcctcacggc ccgggagtgg gggtgactgg cgcagcaact tctctcgttc
      361 ctggagagag gccacaatgt ggcgcgacag attgtcgtaa cggactggtg agggctctcg
      421 gtgtgttggg ccagttggca ccaaacgtgg gtgcctctca gcttcccgtt gctgggccag
      481 cctggctgac aggaagggag aggtatagcc taaaggtggg tctggctcag gccctgcctg
      541 cactgactca aaatcagggc tgtctgaagg tgtgagcaag ctgtcataag ataggcttcc
      601 attgcgtgtc tggttggcca ggctcttata ggaggtggag gtggtgccct ctgaacgaat
      661 ggattgcaac tggcccagct caaagcctgt gccctgggct gacttgaggc tggaggagcg
      721 tgagccactg gatagtggat cgaagtgaaa acttttgcca aaggtaggag aacggaagct
      781 ctctggttcc aagctgggct ctgagcggta gctggggtga cggctgctct ctgcaattga
      841 ggttggctcc ttcaagctgt ccccacgact caacttggcg ctggaggaat gcggcatggc
      901 agctgacgta ctgctgctac tgtaacccgg ccgatacttg tacatggtag gtgtcggggg
      961 gctgtccttg gccaataagc tactatcctc attagtagcc aggccgaggt gtgttcgcaa
     1021 ccctgtgtaa cggctcaggt ctggcttagg aggaggtgga ggttcagcat ctgcagactg
     1081 gctctctgtt atctccaggc ttcccttaga ctttgttctc ctcagctctc cctggatgcc
     1141 attatccatg atcttcacag ttatctgccc atctgaaact tctggtcgaa ggaagggagg
     1201 tctgattaca attgtcttct ctttctttgg tctccccaaa tacctgggtg ctggagaact
     1261 gcagagaaca cggctgacat tgttacagca gccattggtg aaggggttca cacctccccg
     1321 gaatttaccc gtaacctgtt cattggttgt gcgtcccctg gccaccagaa ccacgtgaaa
     1381 tcccgtgagg ccagctacag ggatgaagaa taagccagcc acacacatta ctgccattgt
     1441 gacagccgtg cggacccctg agagttcctc tatgtggtag aggacataaa ggaggccaaa
     1501 gccaaacaca cccataatgt gggctgtcag ggaaaggagg aaaaggaaaa aataacggta
     1561 gttccggcga ccaatacagt tattcaccca ggggcagtga tgatcaaatt cctccacaca
     1621 gttgtcacag acactgcagt gggaacatcg agggggacgg taaaagcggc aggtggcaca
     1681 ccatttcatg cgcacctgga tgccctttat ctccactgtt ttgtaaaggg gagctcggaa
     1741 atcatcttcc ttgtcctcat cctcctcagc tcgagggaaa atccctgggt ccatgaaggt
     1801 ggccatgctg aagttggcca acacaaagag aaacataatt gcattgtaga tgggcactgc
     1861 aggtgacaca tacaggctta gtcctggaca cgtaaaggca aagaagagtg tcgtagctcc
     1921 cactaggaag atggcggctg cagagaccgg gacatacttg ctgggtttga atctctttcc
     1981 agactctgcg ggcattgaat tcgcggccgc aagcttgtca tcgtcatcct tgtaatcgat
     2041 gtcatgatct ttataatcac cgtcatggtc tttgtagtcc atggttaatt ctgacggttc
     2101 actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     2161 aacggggcgg ggttattacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     2221 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     2281 cgcccattgg tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     2341 atgtactgcc aagtaggaaa gtcccgtaag gtcatgtact gggcataatg ccaggcgggc
     2401 catttaccgt cattgacgtc aatagggggc ggacttggca tatgatacac ttgatgtact
     2461 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     2521 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     2581 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     2641 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcaacatggc
     2701 ggtcatattg gacatgagcc aatataaatg tacatattat gatatagata caacgtatgc
     2761 aatggccaat agccaatatt gatttatgct atataaccaa tgactaatat ggctaattgc
     2821 caatattgat tcaatgtata gatcagcttg gcactggccg tcgttttaca acgtcgtgac
     2881 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     2941 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     3001 ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3061 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3121 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3181 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3241 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3301 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3361 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3421 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3481 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3541 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3601 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3661 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3721 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3781 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3841 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3901 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3961 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4021 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4081 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4141 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4201 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4261 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4321 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4381 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4441 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4501 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4561 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4621 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4681 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4741 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4801 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4861 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4921 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4981 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5041 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5101 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5161 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5221 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5281 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5341 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5401 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5461 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5521 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5581 tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat ttaatacgac
     5641 tcactatagg gaattaactt tctggttttt cagttcctcg aggagctttt tgcaaaagcc
     5701 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     5761 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     5821 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     5881 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     5941 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     6001 acaccctaac tgacacacat tccacagcca agctggccgc gtacccaatt caacaggcat
     6061 ctactgagtg gacccaacgc atgagaggac agtgccaagc aagcaactca aatgtcccac
     6121 cggttgggca tggccaggta gcctatgctg tgtctggacg tcctcctgct ggtatagtta
     6181 ttttaaaatc agaaggacag ggaagggagc agtggttcac gcctgtaatc ccagcaattt
     6241 gggaggccaa ggtgggtaga tcacctgaga ttaggagttg gagaccagcc tggccaatat
     6301 ggtgaaaccc cgtctctacc aaaaaaacaa aaattagctg agcctggtca tgcatgcctg
     6361 gaatcccaac aactcgggag gctgaggcag gagaatcgct tgaacccagg aggcggagat
     6421 tgcagtgagc caagattgtg ccactgcact ccagcttggt tcccaataga ccccgcaggc
     6481 cctacaggtt gtcttcccaa cttgcccctt gctccatacc acccccctcc accccataat
     6541 attatagaag gacacctagt cagacaaaat gatgcaactt aattttatta ggacaaggct
     6601 ggtgggcact ggagtggcaa cttccagggc caggagaggc actggggagg ggtcacaggg
     6661 atgccacccg ggatcctcta gagtcgactg gtaccgatat cagatcttca caccgaaatc
     6721 tcataggtgg taccaccaac ccctgacacc ttcttgactc ctcccctcgt ggggctactg
     6781 agaggtggct ggcctgggcc aggggaggct gagcctaagc tcttgggctg cccgttggat
     6841 ttgctgtagg tggtcttcag
//
