LOCUS       sequence-407170-        8888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TEK receptor tyrosine kinase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8888)
  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 8888)
  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-TEK-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     548..567
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        642..746
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(747..815,816..1607)
                     /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(965..984)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1778..2366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2267..2286
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2520..2537
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2654..2675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2690..2695,2696..2713,2714..2720)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2728..2744
                     /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)."
     primer_bind     2733..2755
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2752..2768
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2752..2768
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2817..2938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2828..2847)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2882..2901
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3114..3908)
                     /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(3225..3244)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3835..3854
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3975..4304)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3989..4124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4043..4062)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4289..4309
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4318..4746)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4428..4449)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4640..4659
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4792..5016
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5005..5022
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5045..5062)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5072..5113)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(8617..8636)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8618..8636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8681..8884)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(8710..8730)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(8885..8888,1..376)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg
       61 tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt
      121 ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat
      181 tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc
      241 aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg
      301 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
      361 gtcaataatc aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag cgcaaaacgc
      421 ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt tgtagcttaa
      481 attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag atactggctt
      541 aactatgcgg catcagagca gattgtactg agagtgcacc ataggggatc gggagatctc
      601 ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      661 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      721 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      781 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      841 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      901 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      961 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1021 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1081 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1141 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1201 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1261 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1321 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1381 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1441 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1501 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1561 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1621 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1681 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1741 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1801 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1861 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1921 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1981 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2041 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2101 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2161 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2221 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2281 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2341 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2401 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2461 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2521 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     2581 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     2641 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     2701 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     2761 gctatgacca tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag
     2821 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2881 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg
     2941 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     3001 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg
     3061 atggcaggtt gggcgtcgct tggtcggtca tttcgcgaac cccagagtcc cgctcagaag
     3121 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     3181 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     3241 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     3301 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     3361 tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     3421 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     3481 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     3541 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     3601 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     3661 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     3721 tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     3781 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     3841 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     3901 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atccgaaaat ggatatacaa
     3961 gctcccggga gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     4021 aatagctcag aggcagaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     4081 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     4141 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     4201 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     4261 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagaattaat
     4321 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4381 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4441 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4501 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4561 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4621 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4681 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4741 gcgcgtgggg atacccccta gagccccagc tggttctttc cgcctcagaa gccatagagc
     4801 ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc
     4861 ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat
     4921 tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg
     4981 ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag cgggtttaaa
     5041 ctcaatggtg atggtgatga tgaccggtac gcgtagaatc gagaccgagg agagggttag
     5101 ggataggctt accttcgaac cgcgggccct ctagactcga gcggccgcca ctgtgctgga
     5161 tatctgcaga attgtcttga cccttcttgg ccgcttcttc agcagaacag tcaattcctg
     5221 cataagtaaa cttctcataa agcgtggtat tcacgtaggt ctttcgctcc tctaacattc
     5281 tgtttaagga caccaatatc tgggcaaatg atggcctctc ataaggcttc tcccgccagc
     5341 attgtctcat tagatcatac acctcatcat cacagttcag gggcttctcc agtctgtagc
     5401 cctggggcag cttctcgtag agttctgcac aagtcatccc gcagtagggt gtgcctccta
     5461 agctaacaat ctcccatagt aacacaccat aggaccatac atcactgttg gttgtgtaca
     5521 cactgtaatt cagtgactcg atggccatcc agcgcactgg gagccttccc attgtctttt
     5581 tcacatacac ctcttgacct cgggacaatc caaaatctgc tatttttgcc acatagtttt
     5641 caccaactaa aatgtttctg gcagccagat ccctgtggat aaactgtttt tggctcaagt
     5701 agtccatgcc ccgggccacg tcggcagcga agtgaaggag ctgctgggag gacagtgtgg
     5761 acgcggtgct attggcaatg gcaaatgctg ggtccgtctc cagcacacgg ctcttgcgaa
     5821 ggaagtccag aaggtttcca tggggcgcgt actcaatggc caggtacaag tagcctcgat
     5881 gttcacatgc tcctaagaga ttgatgatgt ttggatggtg tccaagttta caaagaactt
     5941 ccagttctcc tgcaaagtcc ctgtgatcat ctttggaggc atattctttc attcttttga
     6001 tggcagcatc catccgtaac ccatccttct tgatgcgcgc cttaagaact tggccaaaat
     6061 tgccctcccc aatcacatct tgaaatttga tgtcattcca gtcaagcact ggataaattg
     6121 taggatctgg gttgtttttg accttcctgt ttagggccag agtccctgag ttgaactgca
     6181 cagctggttc ttccctcacg ttttggaagg cttgggccat tctcctttgc acatttgctc
     6241 tcttcaattg caatatgatc agaaaggcca acagcacagt caggcaggtc attccagcag
     6301 agccaaggat ggctataagc agcatcttcc cccctccgag gtccgctggt gcttgagatt
     6361 ctgggagggt caccagttca tgagaaaagg ctgggttgct tgaccctatg ttgttctctg
     6421 caaaaatgtc cacctggtat gctgtttcag gctctaggcc cttgagctga tactgagtga
     6481 tggtggcatt ctttatcttc acatcaacgt gctggtcttc attcttgcct tgaaccttgt
     6541 aacggatagt aatagaagaa atagaatagc catccaatat tgtccaagaa atcacagccg
     6601 aggagtgtgt aatgttggaa atcttgatgt tttctggttg aggaggaaga atgtcactaa
     6661 gggtccaagc agtgagatct tcactccatt ccccctgggc cttggtgttg actctagctc
     6721 ggaccacgta ctgctccctg ggatgtaagt tgttaagtag caccgaagtc aagttgcctg
     6781 gaactttaat attctgctga tcactttttt gcacagacct tctctccact tcaacataaa
     6841 agtcatcttc cgagcttgga aatattggtt gccaggtcaa atttagagtg gtctgacttt
     6901 taggcaggag atttagacct cttggaggag ggagtccgat agaagctgtt gtgaagcgtc
     6961 tcacaggtcc aggatgccct tccccaccct ctccacgacg gaccagttgc acacagagtt
     7021 catattctgt ccgaggttcc aaatagttga gtgtaacaat ctcatttgtc acttgaatat
     7081 gttgccaagc ctcatagtga ttaacgggtt tgtatagaag cttcttggat ttgattggtc
     7141 catccccaaa gtaaggctca gagctgatgt tgatgacagc aaagttatgt ccagtgtcaa
     7201 tcacgtttgg ggcattcagg ggctttggaa gaactttaac agaaatgttg aagggctttt
     7261 ccaccatccc agccactgtg ttcacactgc agacccaaac tcctgagtca ggggggagga
     7321 tccggtggat ggtgaatatg gctactgaga aatgatccgt atggttaaag tcttttggat
     7381 ggagcactgt cccatccggc ttcaccaggg tcatttcttc attagtaggt agcggccagc
     7441 cagaagcttt gcaaatggga ttaaatttac cactgtttac ttctatatga tctggcaaat
     7501 ccactatctt tggggtcatc ctcggtatgc cttctctctc acactggagc ccctgccatc
     7561 ctggagagca gagacatcct tggaagcgat cacacatctc cccattgttg cagctgcacc
     7621 taagcttaca atctggcccg taaaaaccag ggtggcatgc ttcattgcac tgcagaccct
     7681 tccagcctgt ggcacaggaa cacccatagg ggtcagggag acagaacaca taagacttgc
     7741 atccctcttg tccactgcac ctttctttac aagttctgcc aaacgtgtgc agttcacaag
     7801 ccttctcaca cgtccttccc ataaacccag gagggcaaat gcattctcca gtatcttcat
     7861 ggcagacacc attgttcata caagcagtac agagatggtt gcattcaggt ccccacttct
     7921 gggcttcaca tctccggact atcagcctgg tgaaggccga ggtgaagagg tttcctccta
     7981 tatacctggc cgagtacact ccagcatcct ggggctgagc atgaggcagg tgtacttcta
     8041 gaatatcagg tacttcatgc cggggcactg aatggatgaa ggaaccattt ttgtaaatca
     8101 ctgcatcttc ttctttaatc aatacctttt tgaaagatat gttcacgtta tctcccttgt
     8161 ccacagtcat agttaaagta gctggtagga aggaagcttg ttgacgcatc ttcatggttc
     8221 gtatcctgat tgcctctcct cgaactcgcc cttcacagaa ataagcacca ttgatcttac
     8281 tagccttttc tctcttccaa acaacttttt tagcccattc tctggtcaca tcttgagtaa
     8341 cttccagcgg atcctggtgc tggttcatta aggcttcaaa gtcccttcct atggtgatgg
     8401 gctcatgggg gcgccaccca gaggcaatgc aggtgagaga tgtttcagca tcagatacaa
     8461 gaggtaggga attgatcaag atcaagtcca tggcaccttc cacagttcca gaaaggagca
     8521 agctgactcc acagagaact aagctggcta aagagtccat ggtgaagggt actggatccg
     8581 agctcggtac caagcttaac tagccagctt gggtctccct atagtgagtc gtattaattt
     8641 cgataagcca gtaagcagtg ggttctctag ttagccagag agctctgctt atatagacct
     8701 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     8761 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     8821 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     8881 tcaccatg
//
