LOCUS       sequence-407175-        8318 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of human ROR1 receptor tyrosine kinase in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8318)
  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 8318)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-ROR1-V5/HIS
FEATURES             Location/Qualifiers
     source          1..8318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     207..227
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(236..664)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(346..367)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     558..577
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    710..934
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     923..940
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(963..980)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(990..1031)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3965..3984)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3966..3984)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4029..4232)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4058..4078)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4233..4612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4784..4803
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4878..4982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4983..5051,5052..5843)
                     /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(5201..5220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6014..6602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6503..6522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6756..6773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6890..6911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6926..6931,6932..6949,6950..6956)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6964..6980
                     /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     6969..6991
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6988..7004
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6988..7004
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    7053..7174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7064..7083)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7118..7137
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7350..8144)
                     /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(7461..7480)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8071..8090
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(8211..8318,1..222))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(8225..8318,1..42)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8279..8298)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
       61 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
      121 tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc
      181 tgctggggag cctggggact ttccacaccc taactgacac acattccaca gaattaattc
      241 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
      301 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
      361 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
      421 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
      481 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
      541 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
      601 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
      661 gcgtggggat accccctaga gccccagctg gttctttccg cctcagaagc catagagccc
      721 accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
      781 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
      841 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
      901 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttaaact
      961 caatggtgat ggtgatgatg accggtacgc gtagaatcga gaccgaggag agggttaggg
     1021 ataggcttac cttcgaaccg cgggccctct agactcgagc ggccgccact gtgctggata
     1081 tctgcagaat tgtcttgacc ctttgcagaa atcatagatt cggtgtgtcc atgaatattg
     1141 gcgtctccta gtaaagatgc ttgctttgag tcaattttgt agggtttttg agatttgttg
     1201 ccaaaaacgg tgatacccat tccaccagga tgatttggaa ttgacatgtg tggtagtaaa
     1261 ggaatatttg cttcctgatt ggatcctgat gagggcaagc tagtcacatg gccagtgcta
     1321 gtcgacccac tggcactgct tggggaccga ctcttgggag gtgggcagtg ctgaatcact
     1381 ctgggaggac ctgttggctg gtagtgggca gctggaaacg ctgcatatcc aggaggtatt
     1441 gggtatccat tgatgggaat gaatcgctgg ttctgaggta ttggcgggcc aatgaaacca
     1501 gcaatctggc cctgtggtgt aataccctgg ctcgggaaca tgtaattagg atatctgggg
     1561 ttactgagat tactcactgg gctggcactg agggaggttg tctgtgtggt ggcattcccc
     1621 cccgaaggag tagtagagct tgtgtgactt gagagtccct cccaggaccg aagccggacg
     1681 tgaatatctt taaatcttgg tctcctagaa ggaatctcat tccagcactc tgtcatgagg
     1741 ctgtacattc tgggtgggca gtcttcagag catggtaaga gctgccgttt tctcaccatc
     1801 tcaatcactt cctggttact gaatccataa tatggctgga gtccaaaact gaaaatctcc
     1861 cacaagacaa ccccaaagga ccagatatct gaatcagaag agaatttgcc atacatgatg
     1921 gcttcagggg gcatccagcg aatgggcagc aaggacttac tctggaccct gtagtaatca
     1981 gcggagtaaa tttctctgga aagccccaag tctgaaatct ttacatgaag ttgctctccg
     2041 attaaaatat tgcgagctgc aaggtccttg tggacaaaga agtgactaga caggtattcc
     2101 atgccagctg caatctgaat tgcaatgtgc agaaaatctc cgtggtccag gctggatttc
     2161 acagtcccat cttcatcact gctgcagcca acatcagagt gtggggatct catgatgagg
     2221 aactcatgga gatccccctg attaatatac tcaaaaagca tgcacacagg ttgttcctga
     2281 gtgacggcac ctagaaggca gacaatattg gggtggtgca gttctgccat tagggaggct
     2341 tcttgttgaa attccgtcca ttgctggggg ttgttatagt ctttcaaggt cttgatagca
     2401 accagctgag catggtccat gcctgggaga tagagatggc ctttatagat ttttccaaag
     2461 gcacactcac ccaattcttc cataaagcgt acagcagaaa gaggtagctc tttagccttg
     2521 ctcttgggtt tatatgcatt cagcattgac atctctacat tttgacctct gacgtgtttt
     2581 ggttgcctct ggactggtgc cgacgatgac ttctggttat tccgacagac gcaaatgaag
     2641 aagaagagta aagcaatggc caggggaatg gccacacttg gcactagtat gtacaggatt
     2701 tccattttat tcttctcctt ggaatccttt gaatcgcacg ctgggatgtc acacagatca
     2761 gacttaaagt tttcatccaa ggtgaagcac cagggagctt ccttttgatt ccctgggttg
     2821 cggcagtagg aatggcctcc attcagctct gggaaacgaa gggcggtgaa agtgtgtgtg
     2881 tggggatact gggaattcca tggctggcac tggcgccctg atttggtcac actgacggtc
     2941 ccccggtagt ccacacctgt gctgttataa cacttgtgat ttttatttat aggatctgcc
     3001 atgggaattc caatccggat acagttcgca gcttctgggc tctctggctg ggggagatct
     3061 tcacagtttg gcagtttcag cctcatcaga atcatgggat ttgatcttgc aaaaatgtac
     3121 tctgtttgac acaggacatt ctccaggatt tcacattcat cgcgacacaa gtcacggggc
     3181 tttgggacgg atgaagtttc atcgcagtac gggaaggcat agtggcacag ggaaggaatg
     3241 gcgaactgag aacacttatc agataagtga ctggaagtgc caatcatagt gaaggcagct
     3301 gtgatctgat tttctatttc cccttgcatg tgcaaagact ccatatagac ggtgcggttg
     3361 ccaataaatc ttgcacatgc aatccctctg tatggctgac agaatccatc ttcttcatac
     3421 tcatctgagt atcctggact tgcagtggga ggggggccaa acttgacaaa caagactcca
     3481 gtggaagaaa ccacctcctt gccgtttgtt gccacgcact ggaagtagcc tgtgtctgtg
     3541 gtgtcgaggt ttctaatccg cagccgagag ccatagatgg tggaccgaaa ggagagcctc
     3601 cggggctcct ggaccacagg agcatcattt ttgaaccagc ggatggtggg aggtggattc
     3661 ccagagactt tgcagtgcag ttctgctgtc tggcccagag acgtggtgat gttattcatt
     3721 ggttcatcga gggtcaggta agaatctttg ttgagttcac ttgagatgtt ccatgatgag
     3781 gtaggcacta attcagcact gactgacagc tctgtttctt gggcagcagc cccgcgtgcg
     3841 gccagcagca gcgcggccag cagcgccagg agcggcgggc gcgtcccgcg gcggcgcggt
     3901 cgatgcatgg tgaagggtac tggatccgag ctcggtacca agcttaacta gccagcttgg
     3961 gtctccctat agtgagtcgt attaatttcg ataagccagt aagcagtggg ttctctagtt
     4021 agccagagag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     4081 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     4141 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     4201 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag
     4261 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     4321 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     4381 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     4441 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     4501 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     4561 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcaacgcgta
     4621 tatctggccc gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag attcttcatg
     4681 caattgtcgg tcaagccttg ccttgttgta gcttaaattt tgctcgcgca ctactcagcg
     4741 acctccaaca cacaagcagg gagcagatac tggcttaact atgcggcatc agagcagatt
     4801 gtactgagag tgcaccatag gggatcggga gatctcccga tccgtcgacg tcaggtggca
     4861 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4981 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5041 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5101 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5161 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5221 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5281 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5341 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5401 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5461 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5521 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5581 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5641 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5701 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5761 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5821 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6781 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6841 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6901 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6961 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     7021 tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt ttggacaaac
     7081 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     7141 atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac tccagcatga
     7201 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     7261 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     7321 cggtcatttc gcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
     7381 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     7441 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     7501 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     7561 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
     7621 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     7681 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     7741 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     7801 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     7861 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     7921 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     7981 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     8041 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     8101 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     8161 cctgtctctt gatcagatcc gaaaatggat atacaagctc ccgggagctt tttgcaaaag
     8221 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc agaggcggcc
     8281 tcggcctctg cataaataaa aaaaattagt cagccatg
//
