LOCUS       sequence-462865-        6611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC17protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6611)
  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 6611)
  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-ZDHHC17
FEATURES             Location/Qualifiers
     source          1..6611
                     /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    2952..3574
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3151..3170)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3598..3618)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3603..3932
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3783..3918
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3970..3989)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3971..3989)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4003..4019)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4003..4019)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4016..4038)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4027..4043
                     /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(4051..4057,4058..4075,4076..4081))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4096..4117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4234..4251)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4405..4993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4485..4504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5164..5955,5956..6024))
                     /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     5787..5806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6025..6129)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6156..6584
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6243..6262)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6453..6474
                     /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 ttcaatgcag
     1021 cgggaggagg gatttaacac caagatggcg gacggcccgg atgagtacga taccgaagcg
     1081 ggctgtgtgc cccttctcca cccagaggaa atcaaacccc aaagccatta taaccatgga
     1141 tatggtgaac ctcttggacg gaaaactcat attgatgatt acagcacatg ggacatagtc
     1201 aaggctacac aatatggaat atatgaacgc tgtcgagaat tggtggaagc aggttatgat
     1261 gtacggcaac cggacaaaga aaatgttacc ctcctccatt gggctgccat caataacaga
     1321 atagatttag tcaaatacta tatttcgaaa ggtgctattg tggatcaact tggaggggac
     1381 ctgaattcaa ctccattgca ctgggccaca agacaaggcc atctatccat ggttgtgcaa
     1441 ctaatgaaat atggtgcaga tccttcatta attgatggag aaggatgtag ctgtattcat
     1501 ctggctgctc agttcggaca tacctcaatt gttgcttatc tcatagcaaa aggacaggat
     1561 gtagatatga tggatcagaa tggaatgacg cctttaatgt gggcagcata tagaacacat
     1621 agtgtggatc caactagatt gcttttaaca ttcaatgttt cagttaacct tggtgacaag
     1681 tatcacaaaa acactgctct gcattgggca gtgctagcag ggaataccac agtcattagc
     1741 cttcttctgg aagctggagc taatgttgat gcccagaata tcaagggcga atcagcgctt
     1801 gatttggcaa aacagagaaa aaatgtgtgg atgatcaacc acttacaaga ggcaaggcaa
     1861 gcaaaaggat atgacaatcc gtccttcctt agaaagctga aagctgataa ggaatttcgg
     1921 cagaaagtaa tgttaggaac tcctttccta gttatttggc tggttgggtt tatagcagac
     1981 ctaaatattg attcttggct cattaaaggg ctaatgtatg gtggtgtttg ggctacagta
     2041 cagtttcttt caaaatcctt tttcgatcat tcaatgcata gtgcattgcc ccttgggata
     2101 tatttggcaa ccaaattctg gatgtatgtg acgtggttct tctggttttg gaatgatctc
     2161 aactttttat ttatccatct tccattcctt gccaatagtg ttgcactttt ctacaatttt
     2221 ggaaaatctt ggaaatcaga tccagggatt attaaagcaa cagaagagca aaagaaaaag
     2281 acaatagttg aacttgcaga gacaggaagt ctggacctca gtatattctg cagtacctgt
     2341 ttgatacgaa aaccggtgag gtccaaacat tgtggtgtgt gcaaccgctg tatagcaaaa
     2401 tttgatcatc attgcccatg ggtgggtaac tgtgtaggtg caggcaacca tagatatttt
     2461 atgggctacc tattcttctt gctttttatg atctgctgga tgatttatgg ttgtatatct
     2521 tactggggac tccactgtga gaccacttac accaaggatg gattttggac atacattact
     2581 cagattgcca cgtgttcacc ttggatgttt tggatgttcc tgaacagtgt tttccacttc
     2641 atgtgggtgg ctgtattact catgtgtcag atgtaccaga tatcatgttt aggtattact
     2701 acaaatgaaa gaatgaatgc caggagatac aagcacttta aagtcacaac aacgtctatt
     2761 gaaagcccat tcaaccatgg atgtgtaaga aatattatag acttctttga atttcgatgc
     2821 tgtggcctct ttcgtcctgt tatcgtggac tggaccaggc agtatacaat agaatatgac
     2881 caaatatcag gatctgggta ccagctggtg tagagatctg atatcggtac cagtcgactc
     2941 tagaggatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     3001 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     3061 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3121 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3181 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3241 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3301 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     3361 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     3421 ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc
     3481 tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat
     3541 gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga
     3601 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3661 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3721 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3781 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3841 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3901 gaggcttttt tggaggccta ggcttttgca aaaagctcct cgaggaactg aaaaaccaga
     3961 aagttaattc cctatagtga gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt
     4021 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4081 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4141 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4201 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4261 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4321 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4381 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4441 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4501 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4561 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4621 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4681 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4741 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4801 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4861 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4921 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4981 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5041 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5101 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5161 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5221 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5281 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5341 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5401 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5461 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5521 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5581 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5641 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5701 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5761 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5821 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5881 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5941 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6001 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6061 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6121 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     6181 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6241 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6301 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6361 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6421 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6481 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6541 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6601 gcttacaatt t
//
