LOCUS       sequence-393576-        8083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TYRO3 receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8083)
  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 8083)
  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-TYRO3-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3581..3622
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3632..3649
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3672..3689)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3678..3902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3948..4376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4035..4054)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4245..4266
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4385..4405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4390..4719
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4570..4705
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4632..4651
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4786..5580
                     /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(4840..4859)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5450..5469
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5756..5877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5793..5812)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5847..5866
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5926..5942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5926..5942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5939..5961)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5950..5966
                     /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(5974..5980,5981..5998,5999..6004))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6019..6040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6157..6174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6328..6916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6408..6427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7087..7878,7879..7947))
                     /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     7710..7729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7948..8052)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccagtac ccttcaccat ggcgctgagg cggagcatgg
      961 ggcggcagcg gcgggagccc cggtgaagct gacagtgtct caggggcagc cggtgaagct
     1021 caactgcagt gtggagggga tggaggagcc tgacatccag tgggtgaagg atggggctgt
     1081 ggtccagaac ttggaccagt tgtacatccc agtcagcgag cagcactgga tcggcttcct
     1141 cagcctgaag tcagtggagc gctctgacgc cggccggtac tggtgccagg tggaggatgg
     1201 gggtgaaacc gagatctccc agccagtgtg gctcacggta gaaggtgtgc catttttcac
     1261 agtggagcca aaagatctgg cagtgccacc caatgcccct ttccaactgt cttgtgaggc
     1321 tgtgggtccc cctgaacctg ttaccattgt ctggtggaga ggaactacga agatcggggg
     1381 acccgctccc tctccatctg ttttaaatgt aacaggggtg acccagagca ccatgttttc
     1441 ctgtgaagct cacaacctaa aaggcctggc ctcttctcgc acagccactg ttcaccttca
     1501 agcactgcct gcagccccct tcaacatcac cgtgacaaag ctttccagca gcaacgctag
     1561 tgtggcctgg atgccaggtg ccgatggccg agctctgcta cagtcctgta cagttcaggt
     1621 gacacaggcc ccaggaggct gggaagtcct ggctgttgtg gtccctgtgc ccccctttac
     1681 ctgcctgctc cgggacctgg tgcctgccac caactacagc ctcagggtgc gctgtgccaa
     1741 tgccttgggg ccctctccct atgctgactg ggtgcccttt cagaccaagg gtctagcccc
     1801 agccagcgct ccccaaaacc tccatgccat ccgcacagat tcaggcctca tcttggagtg
     1861 ggaagaagtg atccccgagg cccctttgga aggccccctg ggaccctaca aactgtcctg
     1921 ggttcaagac aatggaaccc aggatgagct gacagtggag gggaccaggg ccaatttgac
     1981 aggctgggat ccccaaaagg acctgatcgt acgtgtgtgc gtctccaatg cagttggctg
     2041 tggaccctgg agtcagccac tggtggtctc ttctcatgac cgtgcaggcc agcagggccc
     2101 tcctcacagc cgcacatcct gggtacctgt ggtccttggt gtgctaacgg ccctggtgac
     2161 ggctgctgcc ctggccctca tcctgcttcg aaagagacgg aaagagacgc ggtttgggca
     2221 agcctttgac agtgtcatgg cccggggaga gccagccgtt cacttccggg cagcccggtc
     2281 cttcaatcga gaaaggcccg agcgcatcga ggccacattg gacagcttgg gcatcagcga
     2341 tgaactaaag gaaaaactgg aggatgtgct catcccagag cagcagttca ccctgggccg
     2401 gatgttgggc aaaggagagt ttggttcagt gcgggaggcc cagctgaagc aagaggatgg
     2461 ctcctttgtg aaagtggctg tgaagatgct gaaagctgac atcattgcct caagcgacat
     2521 tgaagagttc ctcagggaag cagcttgcat gaaggagttt gaccatccac acgtggccaa
     2581 acttgttggg gtaagcctcc ggagcagggc taaaggccgt ctccccatcc ccatggtcat
     2641 cttgcccttc atgaagcatg gggacctgca tgccttcctg ctcgcctccc ggattgggga
     2701 gaaccccttt aacctacccc tccagaccct gatccggttc atggtggaca ttgcctgcgg
     2761 catggagtac ctgagctctc ggaacttcat ccaccgagac ctggctgctc ggaattgcat
     2821 gctggcagag gacatgacag tgtgtgtggc tgacttcgga ctctcccgga agatctacag
     2881 tggggactac tatcgtcaag gctgtgcctc caaactgcct gtcaagtggc tggccctgga
     2941 gagcctggcc gacaacctgt atactgtgca gagtgacgtg tgggcgttcg gggtgaccat
     3001 gtgggagatc atgacacgtg ggcagacgcc atatgctggc atcgaaaacg ctgagattta
     3061 caactacctc attggcggga accgcctgaa acagcctccg gagtgtatgg aggacgtgta
     3121 tgatctcatg taccagtgct ggagtgctga ccccaagcag cgcccgagct ttacttgtct
     3181 gcgaatggaa ctggagaaca tcttgggcca gctgtctgtg ctatctgcca gccaggaccc
     3241 cttatacatc aacatcgaga gagctgagga gcccactgcg ggaggcagcc tggagctacc
     3301 tggcagggat cagccctaca gtggggctgg ggatggcagt ggcatggggg cagtgggtgg
     3361 cactcccagt gactgtcggt acatactcac ccccggaggg ctggctgagc agccagggca
     3421 ggcagagcac cagccagaga gtcccctcaa tgagacacag aggcttttgc tgctgcagca
     3481 agggctactg ccacacagta gctgtaagaa gggtcaagac aattctgcag atatccagca
     3541 cagtggcggc cgctcgagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc
     3601 tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac
     3661 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3721 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3781 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3841 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3901 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3961 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4021 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4081 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4141 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4201 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4261 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4321 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4381 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4441 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4501 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4561 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4621 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4681 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4741 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4801 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4861 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4921 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4981 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5041 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5101 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5161 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5221 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5281 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5341 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5401 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5461 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5521 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5581 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5641 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5701 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5761 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5821 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5881 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5941 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6001 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6061 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6121 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6181 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6241 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6841 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6901 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7021 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7081 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7141 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7201 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7261 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7321 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7381 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7441 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7501 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7561 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7621 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7681 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7741 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7801 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7861 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7921 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7981 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8041 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
