LOCUS       sequence-462834-        5942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC23 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  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 5942)
  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-ZDHHC23
FEATURES             Location/Qualifiers
     source          1..5942
                     /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    2283..2905
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2482..2501)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2929..2949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2934..3263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3114..3249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3301..3320)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3302..3320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3334..3350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3334..3350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3347..3369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3358..3374
                     /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(3382..3388,3389..3406,3407..3412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3427..3448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3565..3582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3736..4324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3816..3835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4495..5286,5287..5355))
                     /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     5118..5137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5356..5460)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5487..5915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5574..5593)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5784..5805
                     /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 ttcaatgaca
     1021 cagaagggca gtatgaagcc tgtgaagaaa aagaaaaccg aagaacctga attggagccc
     1081 ctgtgctgct gcgagtacat agatcggaat ggggaaaaga accacgtggc tacttgtttg
     1141 tgtgattgtc aagatctgga tgaagggtgt gatcgatgga ttacatgtaa atctttacag
     1201 ccagagactt gtgaaagaat catggataca atttctgatc gcctccgaat tccttggctt
     1261 aggggagcca aaaaagtgaa catcagcatc atccctccgc ttgtcctgct gcctgtcttc
     1321 cttcatgtgg cttcctggca tttcctcctg ggggtggtgg ttttgacctc ccttcctgtg
     1381 ctggcactgt ggtactacta cctcactcac agaaggaaag aacagaccct gtttttcctg
     1441 agccttggac tgttctctct gggctacatg tactatgtgt tcctgcagga agtggtcccc
     1501 aaagggcgtg tgggtcccgt tcagctggcg gttcttacct gcgggttatt tctgatactc
     1561 ttagccttgc acagagccaa gaagaatcca ggctacctca gcaatccagc aagcggtgac
     1621 agatctctaa gcagcagcca gctggagtgc ctgagcagaa aagggcagga gaagaccaaa
     1681 gggttccctg gggcagacat gtcgggcagt ctcaacaatc gcacaacaaa ggatgacccc
     1741 aagggctctt ccaagatgcc agctggaagc cccaccaaag cgaaggagga ctggtgtgcc
     1801 aagtgccagc tggtgcgacc agcccgggca tggcactgcc ggatatgtgg catctgtgtg
     1861 aggagaatgg atcatcattg tgtctggata aatagctgcg ttggagaatc aaatcatcaa
     1921 gcatttatac ttgccctttt gatcttcttg ctcacctcgg tgtatgggat cacactgacc
     1981 ttggacacca tttgtagaga cagaagtgtc ttcacagctc ttttctattg tcctggagtt
     2041 tatgcaaatt acagctcggc tctgtccttc acctgcgtgt ggtactctgt gatcatcaca
     2101 gcaggcatgg cctacatctt cctgatccag ctgatcaaca tcagctacaa tgtgactgag
     2161 cgggaagtcc agcaggccct ccgacagaag actgggcgcc ggctcctctg cgggctcatc
     2221 gtggacacag ggttacttgg atgaagatct gatatcggta ccagtcgact ctagaggatc
     2281 ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     2341 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     2401 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     2461 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     2521 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     2581 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     2641 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     2701 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     2761 tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc
     2821 atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt
     2881 ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc
     2941 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3001 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3061 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3121 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3181 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3241 ttggaggcct aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag aaagttaatt
     3301 ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     3361 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3481 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3541 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3601 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4141 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4561 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4621 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4681 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4741 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4801 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4861 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4921 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4981 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5041 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5101 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5161 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5221 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5281 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5341 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5401 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5461 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5521 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5581 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5641 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5701 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5761 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5821 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5881 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5941 tt
//
