LOCUS       sequence-407208-        8729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human RET receptor tyrosine kinase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8729)
  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 8729)
  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: "Elife."; 
            date: "2017-02-15"; volume: "6. pii: e21536. doi"; pages: " 
            10.7554/eLife.21536"
COMMENT     Sequence Label: pcDNA3.1-RET-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        268..471
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     422..442
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     516..535
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        516..534
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3880..3921
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3931..3948
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3971..3988)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3977..4201
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4247..4675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4334..4353)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4544..4565
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4684..4704)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4689..5018
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4869..5004
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4931..4950
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5085..5879
                     /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(5139..5158)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5749..5768
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6055..6176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6092..6111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6146..6165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6225..6241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6225..6241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6238..6260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6249..6265
                     /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(6273..6279,6280..6297,6298..6303))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6318..6339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6456..6473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6627..7215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6707..6726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7386..8177,8178..8246))
                     /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     8009..8028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8247..8351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8426..8445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8617..8729,1..267)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
       61 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      121 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      181 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      241 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      301 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      361 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      421 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      481 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      541 caagctggct agttaagctt ggtaccgagc tcggatccag tacccttcac catggcgaag
      601 gcgacgtccg gtgccgcggg gctgcgtctg ctgttgctgc tgctgctgcc gctgctaggc
      661 aaagtggcat tgggcctcta cttctcgagg gatgcttact gggagaagct gtatgtggac
      721 caggcggccg gcacgccctt gctgtacgtc catgccctgc gggacgcccc tgaggaggtg
      781 cccagcttcc gcctgggcca gcatctctac ggcacgtacc gcacacggct gcatgagaac
      841 aactggatct gcatccagga ggacaccggc ctcctctacc ttaaccggag cctggaccat
      901 agctcctggg agaagctcag tgtccgcaac cgcggctttc ccctgctcac cgtctacctc
      961 aaggtcttcc tgtcacccac atcccttcgt gagggcgagt gccagtggcc aggctgtgcc
     1021 cgcgtatact tctccttctt caacacctcc tttccagcct gcagctccct caagccccgg
     1081 gagctctgct tcccagagac aaggccctcc ttccgcattc gggagaaccg acccccaggc
     1141 accttccacc agttccgcct gctgcctgtg cagttcttgt gccccaacat cagcgtggcc
     1201 tacaggctcc tggagggtga gggtctgccc ttccgctgcg ccccggacag cctggaggtg
     1261 agcacgcgct gggccctgga ccgcgagcag cgggagaagt acgagctggt ggccgtgtgc
     1321 accgtgcacg ccggcgcgcg cgaggaggtg gtgatggtgc ccttcccggt gaccgtgtac
     1381 gacgaggacg actcggcgcc caccttcccc gcgggcgtcg acaccgccag cgccgtggtg
     1441 gagttcaagc ggaaggagga caccgtggtg gccacgctgc gtgtcttcga tgcagacgtg
     1501 gtacctgcat caggggagct ggtgaggcgg tacacaagca cgctgctccc cggggacacc
     1561 tgggcccagc agaccttccg ggtggaacac tggcccaacg agacctcggt ccaggccaac
     1621 ggcagcttcg tgcgggcgac cgtacatgac tataggctgg ttctcaaccg gaacctctcc
     1681 atctcggaga accgcaccat gcagctggcg gtgctggtca atgactcaga cttccagggc
     1741 ccaggagcgg gcgtcctctt gctccacttc aacgtgtcgg tgctgccggt cagcctgcac
     1801 ctgcccagta cctactccct ctccgtgagc aggagggctc gccgatttgc ccagatcggg
     1861 aaagtctgtg tggaaaactg ccaggcattc agtggcatca acgtccagta caagctgcat
     1921 tcctctggtg ccaactgcag cacgctaggg gtggtcacct cagccgagga cacctcgggg
     1981 atcctgtttg tgaatgacac caaggccctg cggcggccca agtgtgccga acttcactac
     2041 atggtggtgg ccaccgacca gcagacctct aggcaggccc aggcccagct gcttgtaaca
     2101 gtggaggggt catatgtggc cgaggaggcg ggctgccccc tgtcctgtgc agtcagcaag
     2161 agacggctgg agtgtgagga gtgtggcggc ctgggctccc caacaggcag gtgtgagtgg
     2221 aggcaaggag atggcaaagg gatcaccagg aacttctcca cctgctctcc cagcaccaag
     2281 acctgccccg acggccactg cgatgttgtg gagacccaag acatcaacat ttgccctcag
     2341 gactgcctcc ggggcagcat tgttggggga cacgagcctg gggagccccg ggggattaaa
     2401 gctggctatg gcacctgcaa ctgcttccct gaggaggaga agtgcttctg cgagcccgaa
     2461 gacatccagg atccactgtg cgacgagctg tgccgcacgg tgatcgcagc cgctgtcctc
     2521 ttctccttca tcgtctcggt gctgctgtct gccttctgca tccactgcta ccacaagttt
     2581 gcccacaagc cacccatctc ctcagctgag atgaccttcc ggaggcccgc ccaggccttc
     2641 ccggtcagct actcctcttc cggtgcccgc cggccctcgc tggactccat ggagaaccag
     2701 gtctccgtgg atgccttcaa gatcctggag gatccaaagt gggaattccc tcggaagaac
     2761 ttggttcttg gaaaaactct aggagaaggc gaatttggaa aagtggtcaa ggcaacggcc
     2821 ttccatctga aaggcagagc agggtacacc acggtggccg tgaagatgct gaaagagaac
     2881 gcctccccga gtgagcttcg agacctgctg tcagagttca acgtcctgaa gcaggtcaac
     2941 cacccacatg tcatcaaatt gtatggggcc tgcagccagg atggcccgct cctcctcatc
     3001 gtggagtacg ccaaatacgg ctccctgcgg ggcttcctcc gcgagagccg caaagtgggg
     3061 cctggctacc tgggcagtgg aggcagccgc aactccagct ccctggacca cccggatgag
     3121 cgggccctca ccatgggcga cctcatctca tttgcctggc agatctcaca ggggatgcag
     3181 tatctggccg agatgaagct cgttcatcgg gacttggcag ccagaaacat cctggtagct
     3241 gaggggcgga agatgaagat ttcggatttc ggcttgtccc gagatgttta tgaagaggat
     3301 tcgtacgtga agaggagcca gggtcggatt ccagttaaat ggatggcaat tgaatccctt
     3361 tttgatcata tctacaccac gcaaagtgat gtatggtctt ttggtgtcct gctgtgggag
     3421 atcgtgaccc tagggggaaa cccctatcct gggattcctc ctgagcggct cttcaacctt
     3481 ctgaagaccg gccaccggat ggagaggcca gacaactgca gcgaggagat gtaccgcctg
     3541 atgctgcaat gctggaagca ggagccggac aaaaggccgg tgtttgcgga catcagcaaa
     3601 gacctggaga agatgatggt taagaggaga gactacttgg accttgcggc gtccactcca
     3661 tctgactccc tgatttatga cgacggcctc tcagaggagg agacaccgct ggtggactgt
     3721 aataatgccc ccctccctcg agccctccct tccacatgga ttgaaaacaa actctatggt
     3781 agaatttccc atgcatttac tagattcaag ggtcaagaca attctgcaga tatccagcac
     3841 agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat ccctaaccct
     3901 ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc
     3961 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4021 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4081 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4141 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4201 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4261 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4321 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4381 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4441 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4501 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4561 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4621 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4681 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4741 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4801 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4861 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4921 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4981 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5041 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5101 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5161 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5221 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5281 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5341 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5401 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5461 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5521 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5581 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5641 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5701 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5761 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5821 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5881 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5941 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6001 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6061 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6121 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6181 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6241 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6301 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6361 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6961 ttatcgccac tggcagcagc cactggtaac aggattagta gagcgaggta tgtaggcggt
     7021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacacgcaga
     7201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7441 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8341 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     8401 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     8461 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     8521 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     8581 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
     8641 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     8701 aacttacggt aaatggcccg cctggctga
//
