LOCUS       sequence-169510-        7659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To rescue the expression of DHHC5 catalytically inactive mutant in 
            the knockdown cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7659)
  AUTHORS   Tian H, Lu JY, Shao C, Huffman KE, Carstens RM, Larsen JE, Girard L,
            Liu H, Rodriguez-Canales J, Frenkel EP, Wistuba II, Minna JD, 
            Hofmann SL
  TITLE     Systematic siRNA Screen Unmasks NSCLC Growth Dependence by 
            Palmitoyltransferase DHHC5.
  JOURNAL   Mol Cancer Res. 2015 Apr;13(4):784-94. doi: 
            10.1158/1541-7786.MCR-14-0608. Epub 2015 Jan 8.
  PUBMED    25573953
REFERENCE   2  (bases 1 to 7659)
  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"; doi: 
            "10.1158/1541-7786.MCR-14-0608"; journalName: "Mol Cancer Res"; 
            date: "2015-04"; volume: "13"; issue: "4"; pages: "784-94"
COMMENT     Sequence Label: pCI-neo-Flag-DHHS
FEATURES             Location/Qualifiers
     source          1..7659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..66
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             149..160
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    2270..2391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2281..2300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2335..2354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2577..3032
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2664..2683)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2874..2895
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3149..3506
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3149..3340
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3172..3192)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3357..3492
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3557..4351
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVFDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3611..3630)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4221..4240
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4415..4463
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4504..4523)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4623..4645
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4683..4701)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4769..4873
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4874..4942,4943..5734)
                     /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(5092..5111)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5905..6493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6394..6413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6704..7083
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7084..7295
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7246..7266
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7292..7316
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          7456..7588
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     7571..7590
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        7633..7651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gactacaaag accatgacgg tgattataaa gatcatgata tcgattacaa ggatgacgat
       61 gacaagcttg gatccatgcc cgcagagtct ggaaagaggt tcaaacccag caagtatgta
      121 ccggtctcag cagccgccat atttctagtg ggagctacaa cactcttctt tgcctttacg
      181 tgtccaggac taagcctgaa tgtgtcacct gcagtgccca tttacaatgc aattatgttt
      241 ctctttgtgc tggccaactt tagcatggcc acctttatgg acccaggaat ttttccccga
      301 gccgaggaag atgaagacaa agaagatgac ttccgagctc ccctttacaa aacagtggag
      361 atcaagggta tccaggtgcg aatgaaatgg tgtgccacct gccgctttta ccgccctcct
      421 agatgctccc actgcagtgt ctgtgacaac tgtgtggagg aatttgacca tcactcccct
      481 tgggtaaaca actgtattgg tcgcaggaac tacagatact tcttcctttt cctcctttcc
      541 ctgacagctc acatcatggg tgtgtttggc tttggcctcc tttatgtcct ctatcacata
      601 gaggaactct caggggtccg cactgctgtc accatggcag tgatgtgtgt ggctggctta
      661 tttttcatcc ctgtagctgg cctcacagga tttcatgtgg tgctggtggc taggggacgc
      721 acaaccaatg aacaggttac aggtaaattc cggggaggtg tgaatccctt caccaatggc
      781 tgctgtaaca acgttagccg tgtcctctgc agttctccag cacccaggta tttggggaga
      841 ccaaagaaag agaagacaat tgtaatcaga cctcctttcc ttcgaccaga agtgtcagat
      901 gggcagataa ctgtgaagat catggataat ggcatccagg gagaattgag gaggactaag
      961 tctaaaggaa gcctagagat aacagagagc cagtctgcag atgcagaacc tccacctcct
     1021 cctaagccgg acctcagccg atacacgggg cttcgaacac acctcagcct ggctactaat
     1081 gaggatagca gtctcttggg caaggacagt ccccctacac ctaccatgta caagtatcga
     1141 ccaggttaca gcagcagtag cacatcagcc gccatgcctc attcctccag cgccaagttg
     1201 agtcgtggag acagcttaaa ggagccaact tcaattgcag atagcagccg ccatcccagc
     1261 taccgctcag aacccagctt ggagccagag agcttccggt ctcctacttt tggcaaaagc
     1321 tttcattttg atccattgtc cagtggctca cgatcctcca gcctcaagtc agctcagggc
     1381 acgggctttg agctgggcca gttgcagtcc attcgttcag agggcaccac ctccacctcc
     1441 tataagagcc tggccaatca gacacgcaat ggaagtctat cttatgacag cctactcact
     1501 ccttcagaca gccctgattt tgagtcagta caggcagggc ctgagccaga cccaccttta
     1561 ggctacacct ctcccttcct gtcagcccgg ctggcccagc aacgggaagc cgagaggcat
     1621 ccacgtttgc tgccaactgg cccaccacac cgagaaccct caccagtccg ttatgacaat
     1681 ctgtctcgcc acattgtggc ctccctccag gaacgagaga agctgctacg acagtcacct
     1741 ccacttgcag gccgtgagga agaaccaggc ttgggagact caggcattca gtcaacacca
     1801 ggctcaggcc atgccccccg tactagttcc tcctcagatg attccaagag atcacccttg
     1861 agtaagactc cactgggacg cccagctgtt ccccgttttg gcaaaccaga tgggctaaga
     1921 agccggggac tagggtcccc tgaaccaggc acaactgccc cctacctggg ccgatcaata
     1981 tcctatagca gccaaaaagc cccatctggt gtctcagaga cagaggaagt agcattgcag
     2041 ccattactga cacccaaaga tgaagtacag ctcaagacca cctacagcaa atccaatggg
     2101 cagcccaaga gtataggctc agcttcccct ggcccaggcc agccacctct cagtagcccc
     2161 acaaggggag gagtcaagaa ggtgtcaggg gtgggtggca ccacttatga gatttctgtg
     2221 tgagcggccg cttcccttta gtgagggtta atgcttcgag cagacatgat aagatacatt
     2281 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2341 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2401 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     2461 taaaacctct acaaatgtgg taaaatccga taaggatcga tccgggctgg cgtaatagcg
     2521 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc
     2581 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2641 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2701 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2761 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2821 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2881 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2941 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3001 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     3061 ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct
     3121 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
     3181 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3241 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3301 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3361 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3421 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3481 tttttggagg cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact
     3541 taaggctaga gccaccatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3601 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3661 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3721 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3781 tccttgcgca gctgtgttcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3841 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3901 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3961 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggcc ttgtcgatca
     4021 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4081 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4141 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4201 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4261 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4321 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4381 caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     4441 tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg attcgcgtat ggtgcactct
     4501 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4561 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4621 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4681 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4741 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4801 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4861 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4921 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4981 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5041 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5101 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5161 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5221 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5281 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5341 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5401 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5461 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5521 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5581 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5641 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5701 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5761 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5821 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5881 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5941 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6001 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6061 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6121 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6181 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6241 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6301 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6361 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6421 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6481 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6541 ttttgctcac atggctcgac agatcttcaa tattggccat tagccatatt attcattggt
     6601 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
     6661 atgtacattt atattggctc atgtccaata tgaccgccat gttggcattg attattgact
     6721 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     6781 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     6841 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     6901 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     6961 agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7021 atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7081 atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga
     7141 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     7201 gactttccaa aatgtcgtaa caactgcgat cgcccgcccc gttgacgcaa atgggcggta
     7261 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcacta
     7321 gaagctttat tgcggtagtt tatcacagtt aaattgctaa cgcagtcagt gcttctgaca
     7381 caacagtctc gaacttaagc tgcagtgact ctcttaaggt agccttgcag aagttggtcg
     7441 tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa
     7501 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact
     7561 gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa
     7621 ggctagagta cttaatacga ctcactatag gctagcatg
//
