LOCUS       sequence-465264-        6275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC22 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  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 6275)
  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-ZDHHC22
FEATURES             Location/Qualifiers
     source          1..6275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(506..1297,1298..1366))
                     /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     1129..1148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1367..1471)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1546..1565)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1737..2116
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2117..2320
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2271..2291
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2365..2384
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2365..2383
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2861..2870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3271..3336
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3368..3385)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3374..3598
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3644..4072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3731..3750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3941..3962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4081..4101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4086..4415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4266..4401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4328..4347
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4482..5276
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4536..4555)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5146..5165
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5450..5571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5487..5506)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5541..5560
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5620..5636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5620..5636)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5633..5655)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5644..5660
                     /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(5668..5674,5675..5692,5693..5698))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5713..5734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5851..5868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6022..6275,1..335))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6102..6121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
       61 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      121 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      181 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      241 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
      361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
      421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
      481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
      541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
      601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
      661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
      721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
      781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
      841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
      901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
      961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     1321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     1381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     1501 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     1561 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     1621 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     1681 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     1741 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1801 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1861 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1921 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1981 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2041 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2101 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2161 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2221 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     2281 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     2341 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
     2401 gcgtttaaac ttaagcttgg taccgagctc ggatccgcca ccatgctggc cctgcggctg
     2461 ctcaacgtgg tggcccccgc ctacttcttg tgcatctccc tggtgacctt cgtgctgcag
     2521 ctcttcctct tcctgcccag catgcgcgag gaccccgcgg ccgcccggct cttctcgccc
     2581 gccctgctcc acggggcgct cttcctattc ctctcggcca acgccctggg caattacgtc
     2641 cttgtcatcc agaactcccc agacgacctg ggggcctgcc agggggcctc ggccaggaag
     2701 actccatgcc cctcacctag cacccacttc tgccgagtgt gcgccagagt caccctgagg
     2761 cacgaccatc actgtttctt caccggcaac tgcatcggca gcaggaacat gcgcaacttc
     2821 gtcctgttct gcctctacac ctccctggcc tgcctctact ccatggtggc cggcgtggcc
     2881 tacatctccg ctgtcctttc catctccttc gcccacccct tggccttcct cacgctcctg
     2941 cccacctcca tcagccagtt cttctccgga gctgtcctgg gttctgaaat gttcgtcatc
     3001 ctcatgctct acctctggtt cgccatcggc ctggcctgcg ccggcttctg ctgccaccag
     3061 ctgctgttga tcctccgcgg gcagacccgc caccaggtgc ggaagggggt ggcagtgagg
     3121 gcccggccct ggcgcaagaa cttacaagag gtcttcggaa agaggtggct gctgggcctg
     3181 ctggtcccca tgttcaatgt cggaagtgag agctccaagc agcaggataa ggaattctgc
     3241 agatatccag cacagtggcg gccgctcgag gactacaaag accatgacgg tgattataaa
     3301 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc
     3361 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3421 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3481 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3541 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3601 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3661 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3721 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3781 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3841 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3901 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3961 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     4021 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4081 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4141 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4201 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4261 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4321 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4381 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4441 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4501 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4561 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4621 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4681 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4741 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4801 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4861 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4921 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4981 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5041 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5101 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5161 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5221 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5281 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5341 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5401 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5461 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5521 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5581 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5641 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5701 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5761 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5821 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5881 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6241 ttcggtgtag gtcgttcgct ccaagctggg ctgtg
//
