LOCUS       sequence-169519-        7659 bp    DNA     circular SYN 14-APR-2025
DEFINITION  To rescue the expression of DHHC5 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 Apr 14, 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-DHHC5
FEATURES             Location/Qualifiers
     source          1..7659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..69
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             152..163
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    2273..2394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2284..2303)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2338..2357
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2580..3035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2667..2686)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2877..2898
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3152..3509
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3152..3343
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3175..3195)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3360..3495
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3560..4354
                     /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(3614..3633)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4224..4243
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4418..4466
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4507..4526)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4626..4648
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4686..4704)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4772..4876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4877..4945,4946..5737)
                     /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(5095..5114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5908..6496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6397..6416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6707..7086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7087..7298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7249..7269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7295..7319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          7459..7591
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     7574..7593
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        7636..7654
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atggactaca aagaccatga cggtgattat aaagatcatg atatcgatta caaggatgac
       61 gatgacaagc ttggatccat gcccgcagag tctggaaaga ggttcaaacc cagcaagtat
      121 gtaccggtct cagcagccgc catatttcta gtgggagcta caacactctt ctttgccttt
      181 acgtgtccag gactaagcct gaatgtgtca cctgcagtgc ccatttacaa tgcaattatg
      241 tttctctttg tgctggccaa ctttagcatg gccaccttta tggacccagg aatttttccc
      301 cgagccgagg aagatgaaga caaagaagat gacttccgag ctccccttta caaaacagtg
      361 gagatcaagg gtatccaggt gcgaatgaaa tggtgtgcca cctgccgctt ttaccgccct
      421 cctagatgct cccactgcag tgtctgtgac aactgtgtgg aggaatttga ccatcactgc
      481 ccttgggtaa acaactgtat tggtcgcagg aactacagat acttcttcct tttcctcctt
      541 tccctgacag ctcacatcat gggtgtgttt ggctttggcc tcctttatgt cctctatcac
      601 atagaggaac tctcaggggt ccgcactgct gtcaccatgg cagtgatgtg tgtggctggc
      661 ttatttttca tccctgtagc tggcctcaca ggatttcatg tggtgctggt ggctagggga
      721 cgcacaacca atgaacaggt tacaggtaaa ttccggggag gtgtgaatcc cttcaccaat
      781 ggctgctgta acaacgttag ccgtgtcctc tgcagttctc cagcacccag gtatttgggg
      841 agaccaaaga aagagaagac aattgtaatc agacctcctt tccttcgacc agaagtgtca
      901 gatgggcaga taactgtgaa gatcatggat aatggcatcc agggagaatt gaggaggact
      961 aagtctaaag gaagcctaga gataacagag agccagtctg cagatgcaga acctccacct
     1021 cctcctaagc cggacctcag ccgatacacg gggcttcgaa cacacctcag cctggctact
     1081 aatgaggata gcagtctctt gggcaaggac agtcccccta cacctaccat gtacaagtat
     1141 cgaccaggtt acagcagcag tagcacatca gccgccatgc ctcattcctc cagcgccaag
     1201 ttgagtcgtg gagacagctt aaaggagcca acttcaattg cagatagcag ccgccatccc
     1261 agctaccgct cagaacccag cttggagcca gagagcttcc ggtctcctac ttttggcaaa
     1321 agctttcatt ttgatccatt gtccagtggc tcacgatcct ccagcctcaa gtcagctcag
     1381 ggcacgggct ttgagctggg ccagttgcag tccattcgtt cagagggcac cacctccacc
     1441 tcctataaga gcctggccaa tcagacacgc aatggaagtc tatcttatga cagcctactc
     1501 actccttcag acagccctga ttttgagtca gtacaggcag ggcctgagcc agacccacct
     1561 ttaggctaca cctctccctt cctgtcagcc cggctggccc agcaacggga agccgagagg
     1621 catccacgtt tgctgccaac tggcccacca caccgagaac cctcaccagt ccgttatgac
     1681 aatctgtctc gccacattgt ggcctccctc caggaacgag agaagctgct acgacagtca
     1741 cctccacttg caggccgtga ggaagaacca ggcttgggag actcaggcat tcagtcaaca
     1801 ccaggctcag gccatgcccc ccgtactagt tcctcctcag atgattccaa gagatcaccc
     1861 ttgagtaaga ctccactggg acgcccagct gttccccgtt ttggcaaacc agatgggcta
     1921 agaagccggg gactagggtc ccctgaacca ggcacaactg ccccctacct gggccgatca
     1981 atatcctata gcagccaaaa agccccatct ggtgtctcag agacagagga agtagcattg
     2041 cagccattac tgacacccaa agatgaagta cagctcaaga ccacctacag caaatccaat
     2101 gggcagccca agagtatagg ctcagcttcc cctggcccag gccagccacc tctcagtagc
     2161 cccacaaggg gaggagtcaa gaaggtgtca ggggtgggtg gcaccactta tgagatttct
     2221 gtgtgagcgg ccgcttccct ttagtgaggg ttaatgcttc gagcagacat gataagatac
     2281 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2341 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2401 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     2461 aagtaaaacc tctacaaatg tggtaaaatc cgataaggat cgatccgggc tggcgtaata
     2521 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     2581 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2641 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2701 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2761 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2821 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2881 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2941 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3001 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     3061 catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa
     3121 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
     3181 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3241 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3301 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3361 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3421 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3481 gcttttttgg aggcctaggc ttttgcaaaa agcttgattc ttctgacaca acagtctcga
     3541 acttaaggct agagccacca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3601 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3661 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3721 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3781 cgttccttgc gcagctgtgt tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3841 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3901 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3961 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gccttgtcga
     4021 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4081 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4141 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4201 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4261 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4321 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4381 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     4441 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggattcgcg tatggtgcac
     4501 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4561 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4621 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4681 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4741 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4801 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4861 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4921 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4981 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5041 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5101 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5161 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5221 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5281 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5341 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5401 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5461 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5521 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5581 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5641 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5701 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5761 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5821 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6121 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6181 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6241 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6301 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6361 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6421 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6481 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6541 gccttttgct cacatggctc gacagatctt caatattggc cattagccat attattcatt
     6601 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     6661 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttggca ttgattattg
     6721 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     6781 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6841 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     6901 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6961 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     7021 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     7081 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     7141 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     7201 cgggactttc caaaatgtcg taacaactgc gatcgcccgc cccgttgacg caaatgggcg
     7261 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatca
     7321 ctagaagctt tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg
     7381 acacaacagt ctcgaactta agctgcagtg actctcttaa ggtagccttg cagaagttgg
     7441 tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag
     7501 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
     7561 actgacatcc actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct
     7621 taaggctaga gtacttaata cgactcacta taggctagc
//
