LOCUS       sequence-462831-        7010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7010)
  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 7010)
  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-ZDHHC8
FEATURES             Location/Qualifiers
     source          1..7010
                     /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    3351..3973
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3550..3569)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3997..4017)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4002..4331
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4182..4317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4369..4388)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4370..4388)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4402..4418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4402..4418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4415..4437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4426..4442
                     /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(4450..4456,4457..4474,4475..4480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4495..4516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4633..4650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4804..5392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4884..4903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5563..6354,6355..6423))
                     /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     6186..6205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6424..6528)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6555..6983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6642..6661)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6852..6873
                     /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 ttcaatgccc
     1021 cgcagccccg ggacgcgcct caaacccgcc aagtacatcc cggtggccac ggccgccgcg
     1081 ctgctggtcg gctccagcac cctcttcttc gtgttcacgt gcccgtggtt gacacgagct
     1141 gtgtccccag ctgttcccgt ctacaatggc atcatcttcc tctttgtcct ggccaacttc
     1201 agcatggcca ctttcatgga ccctggtgtt ttcccccgag cggatgagga tgaggacaag
     1261 gaggacgact tccgggctcc gctgtacaag aacgtggatg tgcgaggtat ccaggtccgc
     1321 atgaagtggt gtgccacgtg ccacttctac cgcccgccgc gctgctccca ctgcagcgtc
     1381 tgtgacaact gtgtagagga ctttgaccac cactgcccct gggtcaacaa ctgcatcggg
     1441 cgtcgaaact atcgctactt cttcctgttc ctgctgtcac tcagtgcaca catggtgggc
     1501 gtcgtggcct tcggcctggt ctacgtgctg aaccacgctg aggggctggg agccgcgcac
     1561 accaccatca ccatggctgt catgtgtgtg gccggcctct tcttcatccc tgtcattggc
     1621 ctcactggct tccatgtggt gctggtcact cgggggcgca ccaccaacga gcaggtgact
     1681 gggaagttcc gcgggggtgt gaaccctttc acccgaggct gctgtgggaa tgtggagcac
     1741 gtgctgtgta gccccctggc gccccggtac gtggtggagc caccccggct gccgctcgcg
     1801 gtgagtttga agccgccttt ccttaggcct gaactcctgg accgagctgc accgctcaag
     1861 gtcaagctta gtgacaacgg gctgaaggct ggcctgggcc gtagcaagtc caagggcagc
     1921 ctggaccggc tggatgagaa gccactggac ttggggccac cactgccccc caagatagag
     1981 gctggcacgt tcagcagtga cctgcagacc ccgcgcccag gcagtgctga gagtgccctg
     2041 tcggtgcaga ggaccagccc cccgacacct gccatgtaca agtttaggcc ggctttcccc
     2101 acgggtccca aggtgccctt ctgtggacca ggcgagcagg ttccaggccc tgattccctg
     2161 accctggggg acgacagcat ccgtagcctg gactttgtgt ccgagccgag cctggacctc
     2221 cctgactatg ggccaggggg cctgcatgca gcctacccgc catccccacc gctcagcgcc
     2281 tctgatgcct tctcgggcgc tttgcgctcc ctgagcctca aggcctcgag ccggcggggc
     2341 ggggatcatg tggccctgca gcccctgcgc tctgaggggg ggccccccac gccccaccgt
     2401 agcatttttg ccccccatgc actgcccaac cgcaacggca gcctgtccta tgacagcctg
     2461 ctcaatcctg gctcgcctgg tggccacgcc tgccctgccc acccagcagt tggcgtggcc
     2521 ggataccact caccctacct gcatcctggg gcaacgggcg acccgccacg gcccctaccc
     2581 cgcagcttca gccccgtgct gggcccccgc ccccgggagc cctcgcctgt gcgctacgac
     2641 aacctgtcca ggaccatcat ggcatccatc caggagcgca aggacaggga ggagcgtgag
     2701 cgcctgctgc gctcccaggc cgactcactc ttcggcgact caggcgtcta tgacgctccc
     2761 agctcctaca gcctgcagca ggccagtgtg ctgtccgagg gcccccgagg tcccgcgctg
     2821 cgctatggct ccagagacga ccttgtggct gggcccggct tcggtggcgc ccgcaaccct
     2881 gccctgcaga cgtcactgtc ctcgctgtcc agctccgtga gccgtgcacc gcggacgtcg
     2941 tcctcctccc tgcaggctga tcaggccagc agcaacgccc cggggccccg gcccagcagt
     3001 ggctcacaca ggtcacctgc acgccagggc ctgccctccc cgcccggcac tccccactca
     3061 ccatcctacg cgggccccaa agctgtcgcc ttcatccaca cggacctccc agagccaccg
     3121 ccctcgctga ccgtgcagag ggaccaccct cagctgaaga ctcccccaag taagcttaat
     3181 gggcagtccc cgggcctggc ccggctggga cctgccaccg gccccccagg gccctctgcc
     3241 agccctacac ggcacacgct ggttaagaag gtgtccggcg tgggtgggac cacctacgag
     3301 atctcggtgt gaagatctga tatcggtacc agtcgactct agaggatccc gggtggcatc
     3361 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     3421 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     3481 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     3541 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     3601 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     3661 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     3721 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     3781 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
     3841 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
     3901 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
     3961 tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg
     4021 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4081 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4141 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4201 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4261 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4321 gcttttgcaa aaagctcctc gaggaactga aaaaccagaa agttaattcc ctatagtgag
     4381 tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4441 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4501 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4561 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4621 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5641 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5701 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5761 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5821 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5881 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5941 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6001 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6061 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6121 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6181 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6241 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6301 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6361 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6421 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6481 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6541 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6601 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6841 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6901 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6961 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
//
