LOCUS       sequence-465263-        6940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC14 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6940)
  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 6940)
  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-ZDHHC14
FEATURES             Location/Qualifiers
     source          1..6940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     420..439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(658..762)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(837..856)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1028..1407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1408..1611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1562..1582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1656..1675
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1656..1674
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3227..3292
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3324..3341)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3330..3554
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3600..4028
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3687..3706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3897..3918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4037..4057)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4042..4371
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4222..4357
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4284..4303
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4438..5232
                     /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(4492..4511)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5102..5121
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5406..5527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5443..5462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5497..5516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5576..5592)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5576..5592)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5589..5611)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5600..5616
                     /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(5624..5630,5631..5648,5649..5654))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5669..5690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5807..5824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5978..6566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6058..6077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6737..6940,1..588,589..657))
                     /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"
ORIGIN
        1 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
       61 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
      121 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
      181 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
      241 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
      301 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
      361 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
      421 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
      481 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
      541 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
      601 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
      661 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
      721 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
      781 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
      841 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
      901 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
      961 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     1021 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     1081 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1141 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1201 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1261 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     1321 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     1381 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     1441 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     1501 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     1561 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     1621 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
     1681 caagctggct agcgtttaaa cttaagcttg gtaccgagct cggatccact agtccagtgt
     1741 ggtggaattc gccaccatgc ctcccggcgg cggcgggccc atgaaagact gcgagtacag
     1801 ccagatcagc acccacagct cctcccccat ggagtcgccc cacaagaaga agaaaatcgc
     1861 ggcccggagg aaatgggagg tgttcccggg aagaaacaag ttcttctgta acgggaggat
     1921 catgatggcc cggcagacgg gcgtcttcta cctgacgctc gtcctcatcc tggtcactag
     1981 cggactcttc ttcgccttcg actgtccgta cctggcggtg aaaatcaccc ctgccatccc
     2041 tgcagtcgct ggcatcctgt tcttctttgt gatggggacc ctgctccgca ccagcttcag
     2101 cgaccccgga gtcctcccac gagccacgcc tgatgaagcc gccgatctgg aaaggcaaat
     2161 agatatcgca aacggcacca gttcaggggg gtaccgcccg cctcccagaa ccaaagaagt
     2221 catcatcaat ggccagaccg tgaaacttaa atactgtttc acctgcaaga ttttccggcc
     2281 ccctcgcgcc tcccattgca gcctttgtga taactgcgta gaacggtttg atcaccactg
     2341 tccctgggta ggcaactgtg tggggaaaag aaactacaga tttttttata tgtttatttt
     2401 atctctgtct tttctgacag tctttatatt tgcattcgtt atcacccacg tcattcttcg
     2461 ttcacagcaa acaggattcc taaatgccct taaggacagt cctgcaagcg tcctggaggc
     2521 tgtggtgtgc ttcttctctg tctggtccat cgttggcctc tcaggattcc acacctactt
     2581 gatcagctcc aaccagacaa caaatgagga cattaaagga tcctggtcaa ataaaagagg
     2641 taaagaaaat tacaatccct acagctacgg aaatatcttt accaactgct gtgttgccct
     2701 gtgtgggccc atctcaccaa gcctgatcga cagaagaggg tacatccagc ccgacacgcc
     2761 gcagccagca gcaccctcca atggcatcac catgtacggg gccacgcagt cacagagtga
     2821 catgtgcgac caagaccagt gcattcagag caccaaattc gttttgcagg ctgcagccac
     2881 gcccctgctg cagagcgagc ccagcctcac cagcgacgag ctgcacctgc ccgggaagcc
     2941 tggcctgggc acgccctgcg ccagcctcac actgggcccg cccacaccgc ccgcctccat
     3001 gcccaacctc gccgaggcca cgctcgcgga cgtgatgccc cggaaagatg agcacatggg
     3061 ccaccagttc ctgacgcccg atgaggcgcc ctcgcccccc aggctactgg cggcgggcag
     3121 ccccctggcg cacagccgca ccatgcacgt gctgggcctg gccagccagg actccctgca
     3181 tgaggactct gtgcgcggcc tggtgaagct cagctccgtg ctcgaggact acaaagacca
     3241 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     3301 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3361 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3421 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3481 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3541 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3661 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3721 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3781 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3901 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3961 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4021 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4141 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4201 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4261 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4321 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4381 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4441 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4501 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4561 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4621 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4681 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4741 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4801 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4861 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4921 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4981 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5041 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5101 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5161 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5221 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5281 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5341 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5401 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5461 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5521 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5581 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5641 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5701 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5761 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5821 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6541 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6661 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6721 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6781 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6841 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6901 atttatcagc aataaaccag ccagccggaa gggccgagcg
//
