LOCUS       sequence-463065-        5507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC21protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5507)
  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 5507)
  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-ZDHHC21
FEATURES             Location/Qualifiers
     source          1..5507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(224..241)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(224..240)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(233..255)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(381..836)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(518..539)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     730..749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        863..967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(968..1036,1037..1828)
                     /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(1186..1205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1999..2587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2488..2507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2741..2758
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2875..2896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2911..2916,2917..2934,2935..2941)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2949..2965
                     /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     2954..2976
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2973..2989
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2973..2989
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3003..3022
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3003..3021
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3060..3389)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3074..3209
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3374..3394
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    3418..4040
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3822..3841
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(4892..4957)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(4987..5190)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5016..5036)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(5191..5507,1..63)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
       61 gtcaacatgg cggtcatatt ggacatgagc caatataaat gtacatatta tgatatagat
      121 acaacgtatg caatggccaa tagccaatat tgatttatgc tatataacca atgactaata
      181 tggctaattg ccaatattga ttcaatgtat agatcagctt ggcactggcc gtcgttttac
      241 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
      301 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
      361 gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
      421 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
      481 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
      541 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
      601 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
      661 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
      721 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
      781 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
      841 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      901 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      961 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1021 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1081 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1141 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1201 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1261 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1321 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1381 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1441 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1501 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1561 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1621 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1681 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1741 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1801 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1861 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1921 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1981 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2041 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2101 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2161 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2221 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2281 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2341 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2401 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2461 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2521 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2581 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2641 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2701 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2761 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2821 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2881 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     2941 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
     3001 tttaatacga ctcactatag ggaattaact ttctggtttt tcagttcctc gaggagcttt
     3061 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     3121 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     3181 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     3241 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     3301 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     3361 ggggactttc cacaccctaa ctgacacaca ttccacagcc aagctggccg cgtacccaat
     3421 tcaacaggca tctactgagt ggacccaacg catgagagga cagtgccaag caagcaactc
     3481 aaatgtccca ccggttgggc atggccaggt agcctatgct gtgtctggac gtcctcctgc
     3541 tggtatagtt attttaaaat cagaaggaca gggaagggag cagtggttca cgcctgtaat
     3601 cccagcaatt tgggaggcca aggtgggtag atcacctgag attaggagtt ggagaccagc
     3661 ctggccaata tggtgaaacc ccgtctctac caaaaaaaca aaaattagct gagcctggtc
     3721 atgcatgcct ggaatcccaa caactcggga ggctgaggca ggagaatcgc ttgaacccag
     3781 gaggcggaga ttgcagtgag ccaagattgt gccactgcac tccagcttgg ttcccaatag
     3841 accccgcagg ccctacaggt tgtcttccca acttgcccct tgctccatac cacccccctc
     3901 caccccataa tattatagaa ggacacctag tcagacaaaa tgatgcaact taattttatt
     3961 aggacaaggc tggtgggcac tggagtggca acttccaggg ccaggagagg cactggggag
     4021 gggtcacagg gatgccaccc gggatcctct agagtcgact ggtaccgata tcagatctga
     4081 catgattggc aaagtggtag ggaactcgca gtggttgcct ctgcctgaaa ggaatgaacc
     4141 acaggatctt ccaacgagtg ccaaaaactt ctgagaaggt ctgctgccat ggctttcggg
     4201 gcctcgatat atcttcacaa cagtttgaca tcttttcaat agatgttgta tctgtgatga
     4261 tgccaattag ttgagtgtaa aagagtccag ttattccaac taacatagta atgcccataa
     4321 aggctgctag tctcattatg gccaattcat gtctaaaaac aaagaggtcc aaattacgct
     4381 tttttagtgg aagaaaatag taatagtggc agaaagaaaa catcagtgcg tagcaagtaa
     4441 gaagttcagt gtagaaacac aactgcagaa agagccaatg attatcttca ccaacacaat
     4501 tgttaatcca tggacagtga tgatccattc tcctcacaca gtggccgcag cggctgcagt
     4561 gatgggaacg ctttggtctc atcaaattac acttgttaca taattcccag aactcccttt
     4621 ctccatgtgg gatcttgggg ttctcaggga gtcttcctgg atcagttatg gaggccctca
     4681 ctaaggcaac cagacagaat atggaaatgc catagaatat tattattaat atgcctggaa
     4741 tatgtccttc ttcatagtga ggaaagagga caattttggg aattaaaaca atattgtata
     4801 accaaacaaa gacaatcaaa cccatgcagc accaaccatg tgggtcaaca acaaagtgaa
     4861 tccggagacc cattgaattc gcggccgcaa gcttgtcatc gtcatccttg taatcgatgt
     4921 catgatcttt ataatcaccg tcatggtctt tgtagtccat ggttaattct gacggttcac
     4981 taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     5041 cggggcgggg ttattacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     5101 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     5161 cccattggtg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     5221 gtactgccaa gtaggaaagt cccgtaaggt catgtactgg gcataatgcc aggcgggcca
     5281 tttaccgtca ttgacgtcaa tagggggcgg acttggcata tgatacactt gatgtactgc
     5341 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     5401 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     5461 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatat
//
