LOCUS       sequence-393632-        8198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human AXL receptor tyrosine kinase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8198)
  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 8198)
  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-AXL-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8198
                     /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             3696..3737
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3747..3764
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3787..3804)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3793..4017
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4063..4491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4150..4169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4360..4381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4500..4520)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4505..4834
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4685..4820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4747..4766
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4901..5695
                     /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(4955..4974)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5565..5584
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5871..5992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5908..5927)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5962..5981
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6041..6057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6041..6057)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6054..6076)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6065..6081
                     /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(6089..6095,6096..6113,6114..6119))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6134..6155
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6272..6289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6443..7031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6523..6542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7202..7993,7994..8062))
                     /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     7825..7844
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8063..8167)
                     /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 ggcgtggcgg tgccccagga
      961 tgggcagggt cccgctggcc tggtgcttgg cgctgtgcgg ctgggcgtgc atggccccca
     1021 ggggcacgca ggctgaagaa agtcccttcg tgggcaaccc agggaatatc acaggtgccc
     1081 ggggactcac gggcaccctt cggtgtcagc tccaggttca gggagagccc cccgaggtac
     1141 attggcttcg ggatggacag atcctggagc tcgcggacag cacccagacc caggtgcccc
     1201 tgggtgagga tgaacaggat gactggatag tggtcagcca gctcagaatc acctccctgc
     1261 agctttccga cacgggacag taccagtgtt tggtgtttct gggacatcag accttcgtgt
     1321 cccagcctgg ctatgttggg ctggagggct tgccttactt cctggaggag cccgaagaca
     1381 ggactgtggc cgccaacacc cccttcaacc tgagctgcca agctcaggga cccccagagc
     1441 ccgtggacct actctggctc caggatgctg tccccctggc cacggctcca ggtcacggcc
     1501 cccagcgcag cctgcatgtt ccagggctga acaagacatc ctctttctcc tgcgaagccc
     1561 ataacgccaa gggggtcacc acatcccgca cagccaccat cacagtgctc ccccagcagc
     1621 cccgtaacct ccacctggtc tcccgccaac ccacggagct ggaggtggct tggactccag
     1681 gcctgagcgg catctacccc ctgacccact gcaccctgca ggctgtgctg tcagacgatg
     1741 ggatgggcat ccaggcggga gaaccagacc ccccagagga gcccctcacc tcgcaagcat
     1801 ccgtgccccc ccatcagctt cggctaggca gcctccatcc tcacacccct tatcacatcc
     1861 gcgtggcatg caccagcagc cagggcccct catcctggac ccactggctt cctgtggaga
     1921 cgccggaggg agtgcccctg ggcccccctg agaacattag tgctacgcgg aatgggagcc
     1981 aggccttcgt gcattggcaa gagccccggg cgcccctgca gggtaccctg ttagggtacc
     2041 ggctggcgta tcaaggccag gacaccccag aggtgctaat ggacataggg ctaaggcaag
     2101 aggtgaccct ggagctgcag ggggacgggt ctgtgtccaa tctgacagtg tgtgtggcag
     2161 cctacactgc tgctggggat ggaccctgga gcctcccagt acccctggag gcctggcgcc
     2221 cagggcaagc acagccagtc caccagctgg tgaaggaacc ttcaactcct gccttctcgt
     2281 ggccctggtg gtatgtactg ctaggagcag tcgtggccgc tgcctgtgtc ctcatcttgg
     2341 ctctcttcct tgtccaccgg cgaaagaagg agacccgtta tggagaagtg tttgaaccaa
     2401 cagtggaaag aggtgaactg gtagtcaggt accgcgtgcg caagtcctac agtcgtcgga
     2461 ccactgaagc taccttgaac agcttgggca tcagtgaaga gctgaaggag aagctgcggg
     2521 atgtgatggt ggaccggcac aaggtggccc tggggaagac tctgggagag ggagagtttg
     2581 gagctgtgat ggaaggccag ctcaaccagg acgactccat cctcaaggtg gctgtgaaga
     2641 cgatgaagat tgccatctgc acgaggtcag agctggagga tttcctgagt gaagcggtct
     2701 gcatgaagga atttgaccat cccaacgtca tgaggctcat cggtgtctgt ttccagggtt
     2761 ctgaacgaga gagcttccca gcacctgtgg tcatcttacc tttcatgaaa catggagacc
     2821 tacacagctt cctcctctat tcccggctcg gggaccagcc agtgtacctg cccactcaga
     2881 tgctagtgaa gttcatggca gacatcgcca gtggcatgga gtatctgagt accaagagat
     2941 tcatacaccg ggacctggcg gccaggaact gcatgctgaa tgagaacatg tccgtgtgtg
     3001 tggcggactt cgggctctcc aagaagatct acaatgggga ctactaccgc cagggacgta
     3061 tcgccaagat gccagtcaag tggattgcca ttgagagtct agctgaccgt gtctacacca
     3121 gcaagagcga tgtgtggtcc ttcggggtga caatgtggga gattgccaca agaggccaaa
     3181 ccccatatcc gggcgtggag aacagcgaga tttatgacta tctgcgccag ggaaatcgcc
     3241 tgaagcagcc tgcggactgt ctggatggac tgtatgcctt gatgtcgcgg tgctgggagc
     3301 taaatcccca ggaccggcca agttttacag agctgcggga agatttggag aacacactga
     3361 aggccttgcc tcctgcccag gagcctgacg aaatcctcta tgtcaacatg gatgagggtg
     3421 gaggttatcc tgaaccccct ggagctgcag gaggagctga ccccccaacc cagccagacc
     3481 ctaaggattc ctgtagctgc ctcactgcgg ctgaggtcca tcctgctgga cgctatgtcc
     3541 tctgcccttc cacaacccct agccccgctc agcctgctga taggggctcc ccagcagccc
     3601 cagggcagga ggatggtgcc aaaaagggtc aagacaattc tgcagatatc cagcacagtg
     3661 gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     3721 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     3781 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3841 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3901 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3961 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     4021 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     4081 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4141 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4201 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4261 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4321 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4381 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4441 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4501 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4561 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4621 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4681 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4741 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4801 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4861 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4921 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4981 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5041 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     5101 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5161 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5221 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5281 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5341 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5401 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5461 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5521 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5581 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5641 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5701 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5761 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5821 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5881 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5941 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6001 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6061 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6121 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6181 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6241 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6301 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6361 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6421 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6481 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6541 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6601 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6661 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6721 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6781 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6841 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6901 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6961 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7021 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7081 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7141 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7201 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7261 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7321 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7381 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7441 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7501 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7561 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7621 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7681 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7741 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7801 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7861 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7921 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7981 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8041 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8101 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8161 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
