LOCUS       sequence-396819-        8429 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expression of human CSF1R receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8429)
  AUTHORS   Gudernova I, Foldynova-Trantirkova S, Ghannamova BE, Fafilek B, 
            Varecha M, Balek L, Hruba E, Jonatova L, Jelinkova I, Kunova 
            Bosakova M, Trantirek L, Mayer J, Krejci P
  TITLE     One reporter for in-cell activity profiling of majority of protein 
            kinase oncogenes.
  JOURNAL   Elife. 2017 Feb 15;6. pii: e21536. doi: 10.7554/eLife.21536.
  PUBMED    28199182
REFERENCE   2  (bases 1 to 8429)
  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"; journalName: "Elife."; 
            date: "2017-02-15"; volume: "6. pii: e21536. doi"; pages: " 
            10.7554/eLife.21536"
COMMENT     Sequence Label: pcDNA3.1-CSF1R-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..56)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        228..607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        608..811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     762..782
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     856..875
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        856..874
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3920..3961
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3971..3988
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4011..4028)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4017..4241
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4287..4715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4374..4393)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4584..4605
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4724..4744)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4729..5058
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4909..5044
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4971..4990
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5125..5919
                     /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(5179..5198)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5789..5808
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6095..6216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6132..6151)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6186..6205
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6265..6281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6265..6281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6278..6300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6289..6305
                     /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(6313..6319,6320..6337,6338..6343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6358..6379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6496..6513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6667..7255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6747..6766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7426..8217,8218..8286))
                     /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     8049..8068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8287..8391)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
       61 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
      121 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      181 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      241 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      601 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      661 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      721 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      781 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
      841 ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct agttaagctt
      901 ggtaccgagc tcggatccag tacccttcac catgggacca ggagttctgc tgctcctgct
      961 ggtggccaca gcttggcatg gtcagggaat cccagtgata gagcccagtg tccctgagct
     1021 ggtcgtgaag ccaggagcaa cggtgacctt gcgatgtgtg ggcaatggca gcgtggaatg
     1081 ggatggcccc ccatcacctc actggaccct gtactctgat ggctccagca gcatcctcag
     1141 caccaacaac gctaccttcc aaaacacggg gacctatcgc tgcactgagc ctggagaccc
     1201 cctgggaggc agcgccgcca tccacctcta tgtcaaagac cctgcccggc cctggaacgt
     1261 gctagcacag gaggtggtcg tgttcgagga ccaggacgca ctactgccct gtctgctcac
     1321 agacccggtg ctggaagcag gcgtctcgct ggtgcgtgtg cgtggccggc ccctcatgcg
     1381 ccacaccaac tactccttct cgccctggca tggcttcacc atccacaggg ccaagttcat
     1441 tcagagccag gactatcaat gcagtgccct gatgggtggc aggaaggtga tgtccatcag
     1501 catccggctg aaagtgcaga aagtcatccc agggccccca gccttgacac tggtgcctgc
     1561 agagctggtg cggattcgag gggaggctgc ccagatcgtg tgctcagcca gcagcgttga
     1621 tgttaacttt gatgtcttcc tccaacacaa caacactaag ctcgcaatcc ctcaacaatc
     1681 tgactttcat aataaccgtt accaaaaagt cctgaccctc aacctcgatc aagtagattt
     1741 ccaacatgcc ggcaactact cctgcgtggc cagcaacgtg cagggcaagc actccacctc
     1801 catgttcttc cgggtggtag agagtgccta cttgaacttg agctctgagc agaacctcat
     1861 ccaggaggtg accgtggggg aggggctcaa cctcaaagtc atggtggagg cctacccagg
     1921 cctgcaaggt tttaactgga cctacctggg acccttttct gaccaccagc ctgagcccaa
     1981 gcttgctaat gctaccacca aggacacata caggcacacc ttcaccctct ctctgccccg
     2041 cctgaagccc tctgaggctg gccgctactc cttcctggcc agaaacccag gaggctggag
     2101 agctctgacg tttgagctca cccttcgata ccccccagag gtaagcgtca tatggacatt
     2161 catcaacggc tctggcaccc ttttgtgtgc tgcctctggg tacccccagc ccaacgtgac
     2221 atggctgcag tgcagtggcc acactgatag gtgtgatgag gcccaagtgc tgcaggtctg
     2281 ggatgaccca taccctgagg tcctgagcca ggagcccttc cacaaggtga cggtgcagag
     2341 cctgctgact gttgagacct tagagcacaa ccaaacctac gagtgcaggg cccacaacag
     2401 cgtggggagt ggctcctggg ccttcatacc catctctgca ggagcccaca cgcatccccc
     2461 ggatgagttc ctcttcacac cagtggtggt cgcctgcatg tccatcatgg ccttgctgct
     2521 gctgctgctc ctgctgctat tgtacaagta taagcagaag cccaagtacc aggtccgctg
     2581 gaagatcatc gagagctatg agggcaacag ttatactttc atcgacccca cgcagctgcc
     2641 ttacaacgag aagtgggagt tcccccggaa caacctgcag tttggtaaga ccctcggagc
     2701 tggagccttt gggaaggtgg tggaggccac ggcctttggt ctgggcaagg aggatgctgt
     2761 cctgaaggtg gctgtgaaga tgctgaagtc cacggcccat gctgatgaga aggaggccct
     2821 catgtccgag ctgaagatca tgagccacct gggccagcac gagaacatcg tcaaccttct
     2881 gggagcctgt acccatggag gccctgtact ggtcatcacg gagtactgtt gctatggcga
     2941 cctgctcaac tttctgcgaa ggaaggctga ggccatgctg ggacccagcc tgagccccgg
     3001 ccaggacccc gagggaggcg tcgactataa gaacatccac ctcgagaaga aatatgtccg
     3061 cagggacagt ggcttctcca gccagggtgt ggacacctat gtggagatga ggcctgtctc
     3121 cacttcttca aatgactcct tctctgagca agacctggac aaggaggatg gacggcccct
     3181 ggagctccgg gacctgcttc acttctccag ccaagtagcc cagggcatgg ccttcctcgc
     3241 ttccaagaat tgcatccacc gggacgtggc agcgcgtaac gtgctgttga ccaatggtca
     3301 tgtggccaag attggggact tcgggctggc tagggacatc atgaatgact ccaactacat
     3361 tgtcaagggc aatgcccgcc tgcctgtgaa gtggatggcc ccagagagca tctttgactg
     3421 tgtctacacg gttcagagcg acgtctggtc ctatggcatc ctcctctggg agatcttctc
     3481 acttgggctg aatccctacc ctggcatcct ggtgaacagc aagttctata aactggtgaa
     3541 ggatggatac caaatggccc agcctgcatt tgccccaaag aatatataca gcatcatgca
     3601 ggcctgctgg gccttggagc ccacccacag acccaccttc cagcagatct gctccttcct
     3661 tcaggagcag gcccaagagg acaggagaga gcgggactat accaatctgc cgagcagcag
     3721 cagaagcggt ggcagcggca gcagcagcag tgagctggag gaggagagct ctagtgagca
     3781 cctgacctgc tgcgagcaag gggatatcgc ccagcccttg ctgcagccca acaactatca
     3841 gttctgcaag ggtcaagaca attctgcaga tatccagcac agtggcggcc gctcgagtct
     3901 agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     3961 gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     4021 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4081 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4141 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4201 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4261 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4321 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4381 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4441 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4501 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4561 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4621 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4681 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4741 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4801 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4861 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4921 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4981 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5041 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     5101 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5161 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5221 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5281 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5341 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5401 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5461 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5521 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5581 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5641 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5701 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5761 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5821 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5881 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     5941 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6001 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6061 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6121 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6181 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6241 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6301 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6361 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6421 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6481 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7081 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7321 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7381 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7441 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7501 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7561 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7621 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7681 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7741 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7801 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7861 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7921 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7981 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8041 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8101 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8161 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8221 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8281 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8341 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8401 ccgaaaagtg ccacctgacg tcgacggat
//
