LOCUS       sequence-337601-        8906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Two-hybrid system that confers spectinomycin resistance in response 
            to substrate phosphorylation. Constitutive expression of c-src 
            kinase, CDC37, and TCPTP(R222M).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8906)
  TITLE     Fox lab plasmids
REFERENCE   2  (bases 1 to 8906)
  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"
COMMENT     Sequence Label: pB2H2*
FEATURES             Location/Qualifiers
     source          1..8906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..39)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        107..211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             212..1027
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             212..1027
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(285..304)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(1073..1090)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1243..1329
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1421..1448
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2280..2303
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2623..2895
                     /codon_start=1
                     /gene="E. coli rpoZ"
                     /product="DNA-directed RNA polymerase subunit omega"
                     /label=RNAP omega
                     /note="promotes RNA polymerase assembly"
                     /translation="MARVTVQDAVEKIGNRFDLVLVAARRARQMQVGGKDPLVPEENDK
                     TTVIALREIEEGLINNQILDVRERQEQQEQEAAELQAVTAIAEGRR"
     RBS             5163..5174
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(6206..6997)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      7686..7908
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             7956..8906
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVKIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 acggcctatt tggcctaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
       61 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
      121 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
      181 aatgcttcaa taatattgaa aaaggaagag tatgagccat attcaacggg aaacgtcttg
      241 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
      301 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
      361 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
      421 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
      481 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
      541 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
      601 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
      661 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
      721 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
      781 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
      841 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
      901 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
      961 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     1021 tttctaaacg ccatgggcat ggctgttttg gcggatgaga gaagattttc agcctgatac
     1081 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc gacagtagcg
     1141 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     1201 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     1261 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     1321 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     1381 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat
     1441 ggcctttttg cgtttctaca aactcgctga ggattctaga gcacagctaa caccacgtcg
     1501 tccctatctg ctgccctagg tctatgagtg gttgctggat aactttacgg gcatgcataa
     1561 ggctcggtat ctatattcag ggagaccaca acggtttccc tctacaaata attttgttta
     1621 acttttacta gagtaaggag gaaaaaaaaa tgagtatcag cagcagggta aaaagcaaaa
     1681 gaattcagct tggacttaac caggctgaac ttgctcaaaa ggtggggact acccagcagt
     1741 ctatagagca gctcgaaaac ggtaaaacta agcgaccacg ctttttacca gaacttgcgt
     1801 cagctcttgg cgtaagtgtt gactggctgc tcaatggcac ctctgattcg aatgttagat
     1861 ttgttgggca cgttgagccc aaagggaaat atccattgat tagcatggtt agagctcgtt
     1921 cgtggtgtga agcttgtgaa ccctacgata tcaaggacat tgatgaatgg tatgacagtg
     1981 acgttaactt attaggcaat ggattctggc tgaaggttga aggtgattcc atgacctcac
     2041 ctgtaggtca aagcatccct gaaggtcata tggtgttagt agatactgga cgggagccag
     2101 tgaatggaag ccttgttgta gccaaactga ctgacgcgaa cgaagcaaca ttcaagaaac
     2161 tggtcataga tggcggtcag aagtacctga aaggcctgaa tccttcatgg cctatgactc
     2221 ctatcaacgg aaactgcaag attatcggtg ttgtcgtgga agcgagggta aaattcgtag
     2281 actacaagga cgacgatgac aagtggtatt ttgggaagat cactcgtcgg gagtccgagc
     2341 ggctgctgct caaccccgaa aacccccggg gaaccttctt ggtccgggag agcgagacgg
     2401 taaaaggtgc ctatgccctc tccgtttctg actttgacaa cgccaagggg ctcaatgtga
     2461 aacactacct gatccgcaag ctggacagcg gcggcttcta catcacctca cgcacacagt
     2521 tcagcagcct gcagcagctg gtggcctact actccaaaca tgctgatggc ttgtgccacc
     2581 gcctgaccaa cgtctgctaa gtgcagtaag gaggaaaaaa aaatggcacg cgtaactgtt
     2641 caggacgctg tagagaaaat tggtaaccgt tttgacctgg tactggtcgc cgcgcgtcgc
     2701 gctcgtcaga tgcaggtagg cggaaaggat ccgctggtac cggaagaaaa cgataaaacc
     2761 actgtaatcg cgctgcgcga aatcgaagaa ggtctgatca acaaccagat cctcgacgtt
     2821 cgcgaacgcc aggaacagca agagcaggaa gccgctgaat tacaagccgt taccgctatt
     2881 gctgaaggtc gtcgtgcagc tgcggaaccg cagtatgaag aaattccgat ttatctgtaa
     2941 taattctaga gcacagctaa caccacgtcg tccctatctg ctgccctagg tctatgagtg
     3001 gttgctggat aactttacgg gcatgcataa ggctcgtata atatattcag ggagaccaca
     3061 acggtttccc tctacaaata attttgttta acttttacta gagcgaaaaa aagtaaggcg
     3121 gtaatccatg ggctccaagc cgcagactca gggcctggcc aaggatgcct gggagatccc
     3181 tcgggagtcg ctgcggctgg aggtcaagct gggccagggc tgctttggcg aggtgtggat
     3241 ggggacctgg aacggtacca ccagggtggc catcaaaacc ctgaagcctg gcacgatgtc
     3301 tccagaggcc ttcctgcagg aggcccaggt catgaagaag ctgaggcatg agaagctggt
     3361 gcagttgtat gctgtggttt cagaggagcc catttacatc gtcacggagt acatgagcaa
     3421 ggggagtttg ctggactttc tcaaggggga gacaggcaag tacctgcggc tgcctcagct
     3481 ggtggacatg gctgctcaga tcgcctcagg catggcgtac gtggagcgga tgaactacgt
     3541 ccaccgggac cttcgtgcag ccaacatcct ggtgggagag aacctggtgt gcaaagtggc
     3601 cgactttggg ctggctcggc tcattgaaga caatgagtac acggcgcggc aaggtgccaa
     3661 attccccatc aagtggacgg ctccagaagc tgccctctat ggccgcttca ccatcaagtc
     3721 ggacgtgtgg tccttcggga tcctgctgac tgagctcacc acaaagggac gggtgcccta
     3781 ccctgggatg gtgaaccgcg aggtgctgga ccaggtggag cggggctacc ggatgccctg
     3841 cccgccggag tgtcccgagt ccctgcacga cctcatgtgc cagtgctggc ggaaggagcc
     3901 tgaggagcgg cccaccttcg agtacctgca ggccttcctg gaggactact tcacgtccac
     3961 cgagccccag taccagcccg gggagaacct ctaagctagc tttaagaagg agatatacca
     4021 tggtggacta cagcgtgtgg gaccacattg aggtgtctga tgatgaagac gagacgcacc
     4081 ccaacatcga cacggccagt ctcttccgct ggcggcatca ggcccgggtg gaacgcatgg
     4141 agcagttcca gaaggagaag gaggaactgg acaggggctg ccgcgagtgc aagcgcaagg
     4201 tggccgagtg ccagaggaaa ctgaaggagc tggaggtggc cgagggcggc aaggcagagc
     4261 tggagcgcct gcaggccgag gcacagcagc tgcgcaagga ggagcggagc tgggagcaga
     4321 agctggagga gatgcgcaag aaggagaaga gcatgccctg gaacgtggac acgctcagca
     4381 aagacggctt cagcaagagc atggtaaata ccaagcccga gaagacggag gaggactcag
     4441 aggaggtgag ggagcagaaa cacaagacct tcgtggaaaa atacgagaaa cagatcaagc
     4501 actttggcat gcttcgccgc tgggatgaca gccaaaagta cctgtcagac aacgtccacc
     4561 tggtgtgcga ggagacagcc aattacctgg tcatttggtg cattgaccta gaggtggagg
     4621 agaaatgtgc actcatggag caggtggccc accagacaat cgtcatgcaa tttatcctgg
     4681 agctggccaa gagcctaaag gtggaccccc gggcctgctt ccggcagttc ttcactaaga
     4741 ttaagacagc cgatcgccag tacatggagg gcttcaacga cgagctggaa gccttcaagg
     4801 agcgtgtgcg gggccgtgcc aagctgcgca tcgagaaggc catgaaggag tacgaggagg
     4861 aggagcgcaa gaagcggctc ggccccggcg gcctggaccc cgtcgaggtc tacgagtccc
     4921 tccctgagga actccagaag tgcttcgatg tgaaggacgt gcagatgctg caggacgcca
     4981 tcagcaagat ggaccccacc gacgcaaagt accacatgca gcgctgcatt gactctggcc
     5041 tctgggtccc caactctaag gccagcgagg ccaaggaggg agaggaggca ggtcctgggg
     5101 acccattact ggaagctgtt cccaagacgg gcgatgagaa ggatgtcagt gtgtaagtgc
     5161 agaaagagga gaaatactag atgcccacca ccatcgagcg ggagttcgaa gagttggata
     5221 ctcagcgtcg ctggcagccg ctgtacttgg aaattcgaaa tgagtcccat gactatcctc
     5281 atagagtggc caagtttcca gaaaacagaa atcgaaacag atacagagat gtaagcccat
     5341 atgatcacag tcgtgttaaa ctgcaaaatg ctgagaatga ttatattaat gccagtttag
     5401 ttgacataga agaggcacaa aggagttaca tcttaacaca gggtccactt cctaacacat
     5461 gctgccattt ctggcttatg gtttggcagc agaagaccaa agcagttgtc atgctgaacc
     5521 gcattgtgga gaaagaatcg gttaaatgtg cacagtactg gccaacagat gaccaagaga
     5581 tgctgtttaa agaaacagga ttcagtgtga agctcttgtc agaagatgtg aagtcgtatt
     5641 atacagtaca tctactacaa ttagaaaata tcaatagtgg tgaaaccaga acaatatctc
     5701 actttcatta tactacctgg ccagattttg gagtccctga atcaccagct tcatttctca
     5761 atttcttgtt taaagtgaga gaatctggct ccttgaaccc tgaccatggg cctgcggtga
     5821 tccactgtag tgcaggcatt gggatgtctg gcaccttctc tctggtagac acttgtcttg
     5881 ttttgatgga aaaaggagat gatattaaca taaaacaagt gttactgaac atgagaaaat
     5941 accgaatggg tcttattcag accccagatc aactgagatt ctcatacatg gctataatag
     6001 aaggagcaaa atgtataaag ggagattcta gtatacagaa acgatggaaa gaactttcta
     6061 aggaagactt atctcctgcc tttgatcatt caccaaacaa aataatgact gaaaaataca
     6121 atgggaacag ataacagtca gtagggccct aaaaaaaaac cccgcccctg acaggggtgg
     6181 ggttttttgc gtacattggc tccgttcatt tgccgactac cttggtgatc tcgcctttca
     6241 cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc
     6301 caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     6361 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     6421 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     6481 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     6541 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     6601 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     6661 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     6721 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     6781 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     6841 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     6901 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     6961 gttgagtcga tacttcggcg atcaccgctt ccctcatttt tttttcctcc ttactgcacc
     7021 tgcaggtcgg tcgacacaac atacgcgccg gaagctttac gagtaaagcc cggggtgcct
     7081 aatgaacaaa ctttcttgta ttttcataca atgtatcttg ttttagggtc cctaagtctc
     7141 cgcagcgcaa cgaaaggccc agtctttcga ctgagccttt cgttgcattt gcccggatgt
     7201 cctcttgacc gtacgacggg taagcctgtt gatgataccg ctgccttact gggtgcatta
     7261 gccagtctga atgacctgtc acgggataat ccgaagtggt cagactggaa aatcagaggg
     7321 caggaactgc tgaacagcaa aaagtcagat agcaccacat agcagacccg ccataaaacg
     7381 ccctgagaag cccgtgacgg gcttttcttg tattatgggt agtttccttg catgaatcca
     7441 taaaaggcgc ctgtagtgcc atttaccccc attcactgcc agagccgtga gcgcagcgaa
     7501 ctgaatgtca cgaaaaagac agcgactcag gtgcctgatg gtcggagaca aaaggaatat
     7561 tcagcgattt gcccgagctt gcgagggtgc tacttaagcc tttagggttt taaggtctgt
     7621 tttgtagagg agcaaacagc gtttgcgaca tccttttgta atactgcgga actgactaaa
     7681 gtagtgagtt atacacaggg ctgggatcta ttctttttat ctttttttat tctttcttta
     7741 ttctataaat tataaccact tgaatataaa caaaaaaaac acacaaaggt ctagcggaat
     7801 ttacagaggg tctagcagaa tttacaagtt ttccagcaaa ggtctagcag aatttacaga
     7861 tacccacaac tcaaaggaaa aggactagta attatcattg actagcccat ctcaattggt
     7921 atagtgatta aaattaccta gaccaattga gatgtatgtc tgaattagtt gttttcaaag
     7981 caaatgaact agcgattagt cgctatgact taacggagca tgaaaccaag ctaattttat
     8041 gctgtgtggc actactcaac cccacgattg aaaaccctac aaggaaagaa cggacggtat
     8101 cgttcactta taaccaatac gctcagatga tgaacatcag tagggaaaat gcttatggtg
     8161 tattagctaa agcaaccaga gagctgatga cgagaactgt gaaaatcagg aatcctttgg
     8221 ttaaaggctt tgagattttc cagtggacaa actatgccaa gttctcaagc gaaaaattag
     8281 aattagtttt tagtgaagag atattgcctt atcttttcca gttaaaaaaa ttcataaaat
     8341 ataatctgga acatgttaag tcttttgaaa acaaatactc tatgaggatt tatgagtggt
     8401 tattaaaaga actaacacaa aagaaaactc acaaggcaaa tatagagatt agccttgatg
     8461 aatttaagtt catgttaatg cttgaaaata actaccatga gtttaaaagg cttaaccaat
     8521 gggttttgaa accaataagt aaagatttaa acacttacag caatatgaaa ttggtggttg
     8581 ataagcgagg ccgcccgact gatacgttga ttttccaagt tgaactagat agacaaatgg
     8641 atctcgtaac cgaacttgag aacaaccaga taaaaatgaa tggtgacaaa ataccaacaa
     8701 ccattacatc agattcctac ctacgtaacg gactaagaaa aacactacac gatgctttaa
     8761 ctgcaaaaat tcagctcacc agttttgagg caaaattttt gagtgacatg caaagtaagc
     8821 atgatctcaa tggttcgttc tcatggctca cgcaaaaaca acgaaccaca ctagagaaca
     8881 tactggctaa atacggaagg atctga
//
