LOCUS       sequence-298506-        6969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  StrepIII-HA-tagged gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6969)
  AUTHORS   Yadav L, Pietila E, Ohman T, Liu X, Mahato AK, Sidorova Y, Lehti K, 
            Saarma M, Varjosalo M
  TITLE     PTPRA Phosphatase Regulates GDNF-Dependent RET Signaling and 
            Inhibits the RET Mutant MEN2A Oncogenic Potential.
  JOURNAL   iScience. 2020 Feb 21;23(2):100871. doi: 10.1016/j.isci.2020.100871.
            Epub 2020 Jan 31.
  PUBMED    32062451
REFERENCE   2  (bases 1 to 6969)
  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: "iScience.";
            date: "2020-02-21"; volume: "23(2):100871. doi"; pages: " 
            10.1016/j.isci.2020.100871. Epub 2020 Jan 31"
COMMENT     Sequence Label: PTPRA_deltaD1-SHC
FEATURES             Location/Qualifiers
     source          1..6969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    44..62
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    65..83
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     93..117
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    260..284
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1890..1914)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1961..1984
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2021..2044
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2075..2101
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2075..2101
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2145..2162)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2151..2375
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2508..2527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2659..2706
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2714..3736
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2714..3736
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    3866..3987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3903..3922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3957..3976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4036..4052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4036..4052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4049..4071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4060..4076
                     /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(4084..4090,4091..4108,4109..4114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4129..4150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4267..4284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4438..5026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4518..4537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5197..5988,5989..6057))
                     /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     5820..5839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6058..6162)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6237..6256)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6428..6807
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6808..6969,1..42)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(6962..6969,1..13)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctccctat cagtgataga
       61 gatctcccta tcagtgatag agatcgtcga cgagctcgtt tagtgaaccg tcagatcgcc
      121 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
      181 cggactctag cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg
      241 tggaattctg cagatatcaa caagtttgta caaaaaagca ggctccacca tggattcctg
      301 gttcattctt gttctgctcg gcagtggtct gatatgtgtc agtgccaaca atgctaccac
      361 agttgcacct tctgtaggaa ttacaagatt aattaactca tcaacggcag aaccagttaa
      421 agaagaggcc aaaacttcaa atccaacttc ttcactaact tctctttctg tggcaccaac
      481 attcagccca aatataactc tgggacccac ctatttaacc actgtcaatt cttcagactc
      541 tgacaatggg accacaagaa cagcaagcac caattctata ggcattacaa tttcaccaaa
      601 tggaacgtgg cttccagata accagttcac ggatgccaga acagaaccct gggaggggaa
      661 ttccagcacc gcagcaacca ctccagaaac tttccctcct tcagatgaga caccaattat
      721 tgcggtgatg gtggccctgt cctctctgct agtgatcgtg tttattatca tagttttgta
      781 catgttaagg tttaagaaat acaagcaagc tgggagccat tccaattctt tccgcttatc
      841 caacggccgc actgaggatg tggagcccca gagtgtgcca cttctggcca gatccccaag
      901 caccaacagg aaatacccac ccctgcccgt ggacaagctg gaagaggaaa ttaaccggag
      961 aatggcagac gacaataagc tctatctcta tggagataca gaactggaag tgacctctct
     1021 agaaacccac ctgcagaaaa tttacaacaa aatcccaggg accagcaaca atggattaga
     1081 ggaggagttt aagaagttaa catcaatcaa aatccagaat gacaagatgc ggactggaaa
     1141 ccttccagcc aacatgaaga agaaccgtgt tttacagatc attccatatg aattcaacag
     1201 agtgatcatt ccagttaagc ggggcgaaga gaatacagac tatgtgaacg catcctttat
     1261 tgatggctac cggcagaagg actcctatat cgccagccag ggccctcttc tccacacaat
     1321 tgaggacttc tggcgaatga tctgggagtg gaaatcctgc tctatcgtga tgctaacaga
     1381 actggaggag agaggccagg agaagtgtgc ccagtactgg ccatctgatg gactggtgtc
     1441 ctatggagat attacagtgg aactgaagaa ggaggaggaa tgtgagagct acaccgtccg
     1501 agacctcctg gtcaccaaca ccagggagaa taagagccgg cagatccggc agttccactt
     1561 ccatggctgg cctgaagtgg gcatccccag tgacggaaag ggcatgatca gcatcatcgc
     1621 cgccgtgcag aagcagcagc agcagtcagg gaaccacccc atcaccgtgc actgcagcgc
     1681 cggggcagga aggacgggga ccttctgtgc cctgagcacc gtcctggagc gtgtgaaagc
     1741 agaggggatt ttggatgtct tccagactgt caagagcctg cggctacaga ggccacacat
     1801 ggtccagaca ctggaacagt atgagttctg ctacaaggtg gtgcaggagt atattgatgc
     1861 attctcagat tatgccaact tcaagttgga cccagctttc ttgtacaaag tggttgatat
     1921 caaggccgcc gacatcacca gcctgtacaa gaaagtgggg tggagccatc cccagttcga
     1981 gaagggcgga ggatctggcg gcggaagcgg cggaggaagc tggtcccacc ctcagtttga
     2041 gaaaggcacc gagctgggca gcaccatggc cagctacccc tacgacgtgc ccgactacgc
     2101 ctgatgactc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2161 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2221 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2281 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2341 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2401 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2461 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2521 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2581 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2641 tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg
     2701 aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     2761 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     2821 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     2881 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     2941 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     3001 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     3061 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     3121 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     3181 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     3241 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     3301 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     3361 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     3421 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     3481 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     3541 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     3601 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     3661 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     3721 gagggcaaag gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag
     3781 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3841 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3901 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3961 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4021 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4081 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4141 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4201 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4261 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4321 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4381 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4441 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4501 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4561 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4621 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4681 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4741 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4801 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4861 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4921 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4981 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5041 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5101 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5161 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5221 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5281 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5341 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5401 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5461 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5521 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5581 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5641 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5701 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5761 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5821 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5881 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5941 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6001 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6061 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6121 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6181 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     6241 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     6301 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     6361 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     6421 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     6481 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6541 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6601 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6661 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     6721 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     6781 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     6841 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     6901 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     6961 acgcaaatg
//
