LOCUS       sequence-308757-        7752 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 7752)
  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 7752)
  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_C433S-SHC
FEATURES             Location/Qualifiers
     source          1..7752
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    802..826
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3215..3239)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3286..3309
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3346..3369
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3400..3426
                     /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             3400..3426
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3470..3487)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3476..3700
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3833..3852)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3984..4031
                     /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             4039..5061
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4039..5061
                     /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    5191..5312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5228..5247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5282..5301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5361..5377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5361..5377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5374..5396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5385..5401
                     /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(5409..5415,5416..5433,5434..5439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5454..5475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5592..5609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5763..6351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5843..5862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6522..7313,7314..7382))
                     /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     7145..7164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7383..7487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7562..7581)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccact agtccagtgt
      781 ggtggaattc tgcagatatc aacaagtttg tacaaaaaag caggctccac catggattcc
      841 tggttcattc ttgttctgct cggcagtggt ctgatatgtg tcagtgccaa caatgctacc
      901 acagttgcac cttctgtagg aattacaaga ttaattaact catcaacggc agaaccagtt
      961 aaagaagagg ccaaaacttc aaatccaact tcttcactaa cttctctttc tgtggcacca
     1021 acattcagcc caaatataac tctgggaccc acctatttaa ccactgtcaa ttcttcagac
     1081 tctgacaatg ggaccacaag aacagcaagc accaattcta taggcattac aatttcacca
     1141 aatggaacgt ggcttccaga taaccagttc acggatgcca gaacagaacc ctgggagggg
     1201 aattccagca ccgcagcaac cactccagaa actttccctc cttcagatga gacaccaatt
     1261 attgcggtga tggtggccct gtcctctctg ctagtgatcg tgtttattat catagttttg
     1321 tacatgttaa ggtttaagaa atacaagcaa gctgggagcc attccaattc tttccgctta
     1381 tccaacggcc gcactgagga tgtggagccc cagagtgtgc cacttctggc cagatcccca
     1441 agcaccaaca ggaaataccc acccctgccc gtggacaagc tggaagagga aattaaccgg
     1501 agaatggcag acgacaataa gctcttcagg gaggaattca acgctctccc tgcatgtcct
     1561 atccaggcca cctgtgaggc tgcttccaag gaggaaaaca aggaaaaaaa tcgatatgta
     1621 aacatcttgc cttatgacca ctctagagtc cacctgacac cggttgaagg ggttccagat
     1681 tctgattaca tcaatgcttc attcatcaac ggttaccaag aaaagaacaa attcattgct
     1741 gcacaaggac caaaagaaga aacggtgaat gatttctggc ggatgatctg ggaacaaaac
     1801 acagccacca tcgtcatggt taccaacctg aaggagagaa aggagtgcaa gtgcgcccag
     1861 tactggccag accaaggctg ctggacctat gggaatattc gggtgtctgt agaggatgtg
     1921 actgtcctgg tggactacac agtacggaag ttctgcatcc agcaggtggg cgacatgacc
     1981 aacagaaagc cacagcgcct catcactcag ttccacttta ccagctggcc agactttggg
     2041 gtgcctttta ccccgatcgg catgctcaag ttcctcaaga aggtgaaggc ctgtaaccct
     2101 cagtatgcag gggccatcgt ggtccactcc agtgcaggtg tagggcgtac aggtaccttt
     2161 gtcgtcattg atgccatgct ggacatgatg catacagaac ggaaggtgga cgtgtatggc
     2221 tttgtgagcc ggatccgggc acagcgctgc cagatggtgc aaaccgatat gcagtatgtc
     2281 ttcatatacc aagcccttct ggagcattat ctctatggag atacagaact ggaagtgacc
     2341 tctctagaaa cccacctgca gaaaatttac aacaaaatcc cagggaccag caacaatgga
     2401 ttagaggagg agtttaagaa gttaacatca atcaaaatcc agaatgacaa gatgcggact
     2461 ggaaaccttc cagccaacat gaagaagaac cgtgttttac agatcattcc atatgaattc
     2521 aacagagtga tcattccagt taagcggggc gaagagaata cagactatgt gaacgcatcc
     2581 tttattgatg gctaccggca gaaggactcc tatatcgcca gccagggccc tcttctccac
     2641 acaattgagg acttctggcg aatgatctgg gagtggaaat cctgctctat cgtgatgcta
     2701 acagaactgg aggagagagg ccaggagaag tgtgcccagt actggccatc tgatggactg
     2761 gtgtcctatg gagatattac agtggaactg aagaaggagg aggaatgtga gagctacacc
     2821 gtccgagacc tcctggtcac caacaccagg gagaataaga gccggcagat ccggcagttc
     2881 cacttccatg gctggcctga agtgggcatc cccagtgacg gaaagggcat gatcagcatc
     2941 atcgccgccg tgcagaagca gcagcagcag tcagggaacc accccatcac cgtgcactgc
     3001 agcgccgggg caggaaggac ggggaccttc tgtgccctga gcaccgtcct ggagcgtgtg
     3061 aaagcagagg ggattttgga tgtcttccag actgtcaaga gcctgcggct acagaggcca
     3121 cacatggtcc agacactgga acagtatgag ttctgctaca aggtggtgca ggagtatatt
     3181 gatgcattct cagattatgc caacttcaag ttggacccag ctttcttgta caaagtggtt
     3241 gatatcaagg ccgccgacat caccagcctg tacaagaaag tggggtggag ccatccccag
     3301 ttcgagaagg gcggaggatc tggcggcgga agcggcggag gaagctggtc ccaccctcag
     3361 tttgagaaag gcaccgagct gggcagcacc atggccagct acccctacga cgtgcccgac
     3421 tacgcctgat gactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     3481 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3541 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3601 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3661 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3721 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3781 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3841 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3901 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3961 tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt
     4021 ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     4081 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     4141 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     4201 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     4261 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     4321 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     4381 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     4441 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     4501 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     4561 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     4621 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     4681 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     4741 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     4801 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     4861 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     4921 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     4981 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     5041 cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat
     5101 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5161 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5221 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5281 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5341 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5401 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5461 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5521 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5581 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5641 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5701 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5761 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5881 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5941 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6001 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6061 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6181 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6361 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6421 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6661 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7381 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7441 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7501 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     7561 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     7621 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     7681 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     7741 gatatacgcg tt
//
