LOCUS       sequence-393565-        8060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human LTK receptor tyrosine kinase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8060)
  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 8060)
  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-LTK-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8060
                     /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             3558..3599
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3609..3626
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3649..3666)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3655..3879
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3925..4353
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4012..4031)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4222..4243
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4362..4382)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4367..4696
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4547..4682
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4609..4628
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4763..5557
                     /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(4817..4836)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5427..5446
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5733..5854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5770..5789)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5824..5843
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5903..5919)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5903..5919)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5916..5938)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5927..5943
                     /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(5951..5957,5958..5975,5976..5981))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5996..6017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6134..6151)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6305..6893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6385..6404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7064..7855,7856..7924))
                     /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     7687..7706
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7925..8029)
                     /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 gatcccacca tgggctgctg gggacagctg ctggtgtggt
      961 tcggagccgc gggcgccatt ctctgctcta gcccggggtc ccaggagact tttctgcggt
     1021 cctcgcccct gccgctggca agtcccagcc cccgggaccc gaaagtcagc gccccgccta
     1081 gtatcttgga gccagcctcc ccgctgaatt ctccgggcac cgaggggtct tggctgtttt
     1141 ctacctgcgg ggccagcggc cggcatgggc ccacacagac acaatgtgac ggggcgtacg
     1201 cggggaccag cgtggtggtg accgtggggg ccgccgggca gctgagaggc gtgcagctgt
     1261 ggcgcgtgcc gggccctggc cagtatctga tctcagccta cggagccgcg ggcggcaaag
     1321 gcgccaagaa ccacctgtcg cgggcgcatg gcgtcttcgt ctcagcaatc ttctccctcg
     1381 gtctcgggga gtcgctgtac atcctggtgg ggcagcaggg agaggacgcc tgtcccggag
     1441 gtagcccgga gagccagctc gtctgcctcg gggagtctcg agccgttgaa gagcacgcgg
     1501 cgatggatgg gagcgaaggg gtcccggggt cgcggcgctg ggcgggaggt ggcgggggtg
     1561 gcgggggcgc cacctacgtt ttccgggtgc gcgctggcga gctggaaccg ttgctggtgg
     1621 cggccggagg cggcggtcgg gcctacctga ggccgcggga ccgaggccgg actcaggcct
     1681 cccccgagaa actggagaac cgctcggagg cgcccgggag cggcgggaga ggcggggcgg
     1741 caggtggtgg gggcggctgg acgtcgcggg ctccctctcc gcaggccggc cgctcactgc
     1801 aggagggggc ggagggcggc cagggctgct ccgaggcttg ggcgaccctt ggctgggccg
     1861 cggccggcgg cttcgggggc ggcggcgggg cctgcactgc gggcggaggc ggcggcggct
     1921 acaggggggg cgacgcttca gagactgaca acctctgggc tgatggggaa gatggagtat
     1981 ccttcataca ccccagcagc gagctcttcc tgcagcctct ggcagtcacc gagaaccacg
     2041 gagaggtaga gatccgaagg cacctcaact gcagtcactg ccctttgaga gactgccaat
     2101 ggcaggcaga gctccagctg gctgaatgcc tgtgcccaga aggcatggag ctagctgtgg
     2161 ataacgtcac ctgcatggac ctgcacaagc ccccaggccc tctggttctg atggtggctg
     2221 tggtggcaac ctcaacactg agcctcctta tggtgtgtgg ggtcctgatt ctggtgaagc
     2281 agaagaagtg gcagggcctg caggagatga ggctgccgag ccctgagctt gagctgagca
     2341 agcttcgaac ctctgccatc aggacagccc ccaatcccta ttattgccag gtggggcttg
     2401 gcccggccca gtcctggcct ctgccaccag gtgtcaccga ggtttcccca gccaatgtta
     2461 ctctgctcag agccctgggc catggtgcct ttggggaggt gtatgaggga ctggtaattg
     2521 gccttcctgg ggactccagt cccctgcagg tagctatcaa gaccctgcca gaactctgct
     2581 cgcctcagga tgagctggat ttcctcatgg aggccctcat catcagcaag tttcgccatc
     2641 agaacattgt gcggtgtgtg gggctcagcc tcagggccac ccctcgcctc attctgctgg
     2701 aactgatgtc tggaggggac atgaagagtt tcctgaggca cagtcggcca cacctgggcc
     2761 agccatcacc tctggtcatg cgggacctgc tgcaactggc ccaggacata gcccagggct
     2821 gccactacct ggaggaaaat cacttcatcc acagggatat tgccgcccgg aactgcctgc
     2881 tgagctgcgc tggacccagc cgagtggcca agattgggga ctttgggatg gcacgagata
     2941 tctaccgggc cagttattac cgcagggggg accgggcctt gctcccagtc aagtggatgc
     3001 ccccagaggc cttcctggag ggcatcttca catccaagac agattcctgg tcttttgggg
     3061 tgctgctctg ggagatcttc tcactgggct acatgcccta tcctgggcgc accaaccagg
     3121 aggtgctgga cttcgtcgtt ggaggaggcc ggatggaccc tcctaggggc tgcccagggc
     3181 ctgtgtaccg catcatgacc cagtgttggc agcacgagcc tgagctccgc cctagctttg
     3241 ccagcatctt ggagcgtctg cagtactgca ctcaggaccc ggatgtgctg aattcactcc
     3301 tgccaatgga gctggggccc accccagagg aggaagggac ttctgggctg gggaacagat
     3361 ctttggagtg cctaagaccc ccacagcccc aggaactgag tccagagaag ttgaaaagct
     3421 ggggaggtag ccctcttggc ccctggctgt cctctggcct caagcccctc aaatccaggg
     3481 gcctccaacc tcagaacctt tggaatccca cttatcgctc cggcggccgc tcgagtctag
     3541 agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     3601 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     3661 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3721 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3781 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3841 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3901 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3961 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4021 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4081 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4141 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4201 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4261 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4321 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4381 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4441 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4501 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4561 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4621 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4681 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4741 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4801 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4861 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4921 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4981 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5041 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5101 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5161 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5221 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5281 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5341 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5401 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5461 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5521 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     5581 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5641 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5701 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5761 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5821 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5881 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5941 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6001 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6061 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6121 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6181 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6241 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6301 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6361 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6421 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6481 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6541 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6601 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6661 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6721 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6781 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6841 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6901 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6961 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7021 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7081 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7141 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7201 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7261 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7321 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7381 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7441 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7501 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7561 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7621 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7681 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7741 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7801 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7861 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7921 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7981 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8041 gaaaagtgcc acctgacgtc
//
