LOCUS       sequence-270457-        7752 bp    DNA     circular SYN 14-APR-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 Apr 14, 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-SHC
FEATURES             Location/Qualifiers
     source          1..7752
                     /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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1036..1060
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3449..3473)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3520..3543
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3580..3603
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3634..3660
                     /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             3634..3660
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3704..3721)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3710..3934
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4067..4086)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4218..4265
                     /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             4273..5295
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4273..5295
                     /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    5425..5546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5462..5481)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5516..5535
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5595..5611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5595..5611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5608..5630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5619..5635
                     /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(5643..5649,5650..5667,5668..5673))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5688..5709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5826..5843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5997..6585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6077..6096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6756..7547,7548..7616))
                     /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     7379..7398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7617..7721)
                     /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 ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatcaacaag tttgtacaaa aaagcaggct ccaccatgga ttcctggttc
     1081 attcttgttc tgctcggcag tggtctgata tgtgtcagtg ccaacaatgc taccacagtt
     1141 gcaccttctg taggaattac aagattaatt aactcatcaa cggcagaacc agttaaagaa
     1201 gaggccaaaa cttcaaatcc aacttcttca ctaacttctc tttctgtggc accaacattc
     1261 agcccaaata taactctggg acccacctat ttaaccactg tcaattcttc agactctgac
     1321 aatgggacca caagaacagc aagcaccaat tctataggca ttacaatttc accaaatgga
     1381 acgtggcttc cagataacca gttcacggat gccagaacag aaccctggga ggggaattcc
     1441 agcaccgcag caaccactcc agaaactttc cctccttcag atgagacacc aattattgcg
     1501 gtgatggtgg ccctgtcctc tctgctagtg atcgtgttta ttatcatagt tttgtacatg
     1561 ttaaggttta agaaatacaa gcaagctggg agccattcca attctttccg cttatccaac
     1621 ggccgcactg aggatgtgga gccccagagt gtgccacttc tggccagatc cccaagcacc
     1681 aacaggaaat acccacccct gcccgtggac aagctggaag aggaaattaa ccggagaatg
     1741 gcagacgaca ataagctctt cagggaggaa ttcaacgctc tccctgcatg tcctatccag
     1801 gccacctgtg aggctgcttc caaggaggaa aacaaggaaa aaaatcgata tgtaaacatc
     1861 ttgccttatg accactctag agtccacctg acaccggttg aaggggttcc agattctgat
     1921 tacatcaatg cttcattcat caacggttac caagaaaaga acaaattcat tgctgcacaa
     1981 ggaccaaaag aagaaacggt gaatgatttc tggcggatga tctgggaaca aaacacagcc
     2041 accatcgtca tggttaccaa cctgaaggag agaaaggagt gcaagtgcgc ccagtactgg
     2101 ccagaccaag gctgctggac ctatgggaat attcgggtgt ctgtagagga tgtgactgtc
     2161 ctggtggact acacagtacg gaagttctgc atccagcagg tgggcgacat gaccaacaga
     2221 aagccacagc gcctcatcac tcagttccac tttaccagct ggccagactt tggggtgcct
     2281 tttaccccga tcggcatgct caagttcctc aagaaggtga aggcctgtaa ccctcagtat
     2341 gcaggggcca tcgtggtcca ctgcagtgca ggtgtagggc gtacaggtac ctttgtcgtc
     2401 attgatgcca tgctggacat gatgcataca gaacggaagg tggacgtgta tggctttgtg
     2461 agccggatcc gggcacagcg ctgccagatg gtgcaaaccg atatgcagta tgtcttcata
     2521 taccaagccc ttctggagca ttatctctat ggagatacag aactggaagt gacctctcta
     2581 gaaacccacc tgcagaaaat ttacaacaaa atcccaggga ccagcaacaa tggattagag
     2641 gaggagttta agaagttaac atcaatcaaa atccagaatg acaagatgcg gactggaaac
     2701 cttccagcca acatgaagaa gaaccgtgtt ttacagatca ttccatatga attcaacaga
     2761 gtgatcattc cagttaagcg gggcgaagag aatacagact atgtgaacgc atcctttatt
     2821 gatggctacc ggcagaagga ctcctatatc gccagccagg gccctcttct ccacacaatt
     2881 gaggacttct ggcgaatgat ctgggagtgg aaatcctgct ctatcgtgat gctaacagaa
     2941 ctggaggaga gaggccagga gaagtgtgcc cagtactggc catctgatgg actggtgtcc
     3001 tatggagata ttacagtgga actgaagaag gaggaggaat gtgagagcta caccgtccga
     3061 gacctcctgg tcaccaacac cagggagaat aagagccggc agatccggca gttccacttc
     3121 catggctggc ctgaagtggg catccccagt gacggaaagg gcatgatcag catcatcgcc
     3181 gccgtgcaga agcagcagca gcagtcaggg aaccacccca tcaccgtgca ctgcagcgcc
     3241 ggggcaggaa ggacggggac cttctgtgcc ctgagcaccg tcctggagcg tgtgaaagca
     3301 gaggggattt tggatgtctt ccagactgtc aagagcctgc ggctacagag gccacacatg
     3361 gtccagacac tggaacagta tgagttctgc tacaaggtgg tgcaggagta tattgatgca
     3421 ttctcagatt atgccaactt caagttggac ccagctttct tgtacaaagt ggttgatatc
     3481 aaggccgccg acatcaccag cctgtacaag aaagtggggt ggagccatcc ccagttcgag
     3541 aagggcggag gatctggcgg cggaagcggc ggaggaagct ggtcccaccc tcagtttgag
     3601 aaaggcaccg agctgggcag caccatggcc agctacccct acgacgtgcc cgactacgcc
     3661 tgatgactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3721 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3781 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3841 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3901 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3961 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4021 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4081 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4141 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4201 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga
     4261 acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     4321 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     4381 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     4441 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     4501 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     4561 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     4621 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     4681 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     4741 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     4801 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     4861 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     4921 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     4981 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     5041 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     5101 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     5161 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     5221 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     5281 agggcaaagg aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5341 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5401 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5461 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5521 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5581 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5641 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5701 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5761 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5821 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5881 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5941 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6001 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6061 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6121 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6181 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6241 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6301 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6361 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6421 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6481 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6541 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6601 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6661 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6721 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6781 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6841 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6901 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6961 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7021 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7081 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7141 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7201 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7261 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7321 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7381 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7441 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7501 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7561 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7621 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7681 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7741 ccacctgacg tc
//
