LOCUS       sequence-337598-        8917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inactive two-hybrid system that confers spectinomycin resistance in 
            response to substrate phosphorylation. Constitutive expression of 
            c-src kinase, CDC37, and PTP1B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8917)
  TITLE     Fox lab plasmids
REFERENCE   2  (bases 1 to 8917)
  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: pB2HoptX
FEATURES             Location/Qualifiers
     source          1..8917
                     /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             5162..5173
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(6217..7008)
                     /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      7697..7919
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             7967..8917
                     /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 cagtttgaag aaattccgat ttatctgtaa
     2941 taattctaga gcacagctaa caccacgtcg tccctatctg ctgccctagg tctatgagtg
     3001 gttgctggat aactttacgg gcatgcataa ggctcgtata atatattcag ggagaccaca
     3061 acggtttccc tctacaaata attttgttta acttttacta gagcgaaaaa agtaaggcgg
     3121 taatccatgg gctccaagcc gcagactcag ggcctggcca aggatgcctg ggagatccct
     3181 cgggagtcgc tgcggctgga ggtcaagctg ggccagggct gctttggcga ggtgtggatg
     3241 gggacctgga acggtaccac cagggtggcc atcaaaaccc tgaagcctgg cacgatgtct
     3301 ccagaggcct tcctgcagga ggcccaggtc atgaagaagc tgaggcatga gaagctggtg
     3361 cagttgtatg ctgtggtttc agaggagccc atttacatcg tcacggagta catgagcaag
     3421 gggagtttgc tggactttct caagggggag acaggcaagt acctgcggct gcctcagctg
     3481 gtggacatgg ctgctcagat cgcctcaggc atggcgtacg tggagcggat gaactacgtc
     3541 caccgggacc ttcgtgcagc caacatcctg gtgggagaga acctggtgtg caaagtggcc
     3601 gactttgggc tggctcggct cattgaagac aatgagtaca cggcgcggca aggtgccaaa
     3661 ttccccatca agtggacggc tccagaagct gccctctatg gccgcttcac catcaagtcg
     3721 gacgtgtggt ccttcgggat cctgctgact gagctcacca caaagggacg ggtgccctac
     3781 cctgggatgg tgaaccgcga ggtgctggac caggtggagc ggggctaccg gatgccctgc
     3841 ccgccggagt gtcccgagtc cctgcacgac ctcatgtgcc agtgctggcg gaaggagcct
     3901 gaggagcggc ccaccttcga gtacctgcag gccttcctgg aggactactt cacgtccacc
     3961 gagccccagt accagcccgg ggagaacctc taagctagct ttaagaagga gatataccat
     4021 ggtggactac agcgtgtggg accacattga ggtgtctgat gatgaagacg agacgcaccc
     4081 caacatcgac acggccagtc tcttccgctg gcggcatcag gcccgggtgg aacgcatgga
     4141 gcagttccag aaggagaagg aggaactgga caggggctgc cgcgagtgca agcgcaaggt
     4201 ggccgagtgc cagaggaaac tgaaggagct ggaggtggcc gagggcggca aggcagagct
     4261 ggagcgcctg caggccgagg cacagcagct gcgcaaggag gagcggagct gggagcagaa
     4321 gctggaggag atgcgcaaga aggagaagag catgccctgg aacgtggaca cgctcagcaa
     4381 agacggcttc agcaagagca tggtaaatac caagcccgag aagacggagg aggactcaga
     4441 ggaggtgagg gagcagaaac acaagacctt cgtggaaaaa tacgagaaac agatcaagca
     4501 ctttggcatg cttcgccgct gggatgacag ccaaaagtac ctgtcagaca acgtccacct
     4561 ggtgtgcgag gagacagcca attacctggt catttggtgc attgacctag aggtggagga
     4621 gaaatgtgca ctcatggagc aggtggccca ccagacaatc gtcatgcaat ttatcctgga
     4681 gctggccaag agcctaaagg tggacccccg ggcctgcttc cggcagttct tcactaagat
     4741 taagacagcc gatcgccagt acatggaggg cttcaacgac gagctggaag ccttcaagga
     4801 gcgtgtgcgg ggccgtgcca agctgcgcat cgagaaggcc atgaaggagt acgaggagga
     4861 ggagcgcaag aagcggctcg gccccggcgg cctggacccc gtcgaggtct acgagtccct
     4921 ccctgaggaa ctccagaagt gcttcgatgt gaaggacgtg cagatgctgc aggacgccat
     4981 cagcaagatg gaccccaccg acgcaaagta ccacatgcag cgctgcattg actctggcct
     5041 ctgggtcccc aactctaagg ccagcgaggc caaggaggga gaggaggcag gtcctgggga
     5101 cccattactg gaagctgttc ccaagacggg cgatgagaag gatgtcagtg tgtaagtgca
     5161 gaaagaggag aaatactaga tggagatgga aaaggagttc gagcagatcg acaagtccgg
     5221 gagctgggcg gccatttacc aggatatccg acatgaagcc agtgacttcc catgtagagt
     5281 ggccaagctt cctaagaaca aaaaccgaaa taggtacaga gacgtcagtc cctttgacca
     5341 tagtcggatt aaactacatc aagaagataa tgactatatc aacgctagtt tgataaaaat
     5401 ggaagaagcc caaaggagtt acattcttac ccagggccct ttgcctaaca catgcggtca
     5461 cttttgggag atggtgtggg agcagaaaag caggggtgtc gtcatgctca acagagtgat
     5521 ggagaaaggt tcgttaaaat gcgcacaata ctggccacaa aaagaagaaa aagagatgat
     5581 ctttgaagac acaaatttga aattaacatt gatctctgaa gatatcaagt catattatac
     5641 agtgcgacag ctagaattgg aaaaccttac aacccaagaa actcgcgaga tcttacattt
     5701 ccactatacc acatggcctg actttggagt ccctgaatca ccagcctcat tcttgaactt
     5761 tcttttcaaa gtccgagagt cagggtcact cagcccggag cacgggcccg ttgtggtgca
     5821 ctgcagtgca ggcatcggca ggtctggaac cttctgtctg gctgatacct gcctcttgct
     5881 gatggacaag aggaaagacc cttcttccgt tgatatcaag aaagtgctgt tagaaatgag
     5941 gaagtttcgg atggggctga tccagacagc cgaccagctg cgcttctcct acctggctgt
     6001 gatcgaaggt gccaaattca tcatggggga ctcttccgtg caggatcagt ggaaggagct
     6061 ttcccacgag gacctggagc ccccacccga gcatatcccc ccacctcccc ggccacccaa
     6121 acgaatcctg gagccacaca attgacagtc agtagggccc taaaaaaaaa ccccgcccct
     6181 gacaggggtg gggttttttg cgtacattgg ctccgttcat ttgccgacta ccttggtgat
     6241 ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc
     6301 ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg
     6361 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
     6421 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
     6481 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
     6541 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
     6601 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
     6661 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
     6721 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
     6781 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
     6841 tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc
     6901 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
     6961 tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcattt ttttttcctc
     7021 cttactgcac ctgcaggtcg gtcgacacaa catacgcgcc ggaagcttta cgagtaaagc
     7081 ccggggtgcc taatgaacaa actttcttgt attttcatac aatgtatctt gttttagggt
     7141 ccctaagtct ccgcagcgca acgaaaggcc cagtctttcg actgagcctt tcgttgcatt
     7201 tgcccggatg tcctcttgac cgtacgacgg gtaagcctgt tgatgatacc gctgccttac
     7261 tgggtgcatt agccagtctg aatgacctgt cacgggataa tccgaagtgg tcagactgga
     7321 aaatcagagg gcaggaactg ctgaacagca aaaagtcaga tagcaccaca tagcagaccc
     7381 gccataaaac gccctgagaa gcccgtgacg ggcttttctt gtattatggg tagtttcctt
     7441 gcatgaatcc ataaaaggcg cctgtagtgc catttacccc cattcactgc cagagccgtg
     7501 agcgcagcga actgaatgtc acgaaaaaga cagcgactca ggtgcctgat ggtcggagac
     7561 aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc ctttagggtt
     7621 ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac atccttttgt aatactgcgg
     7681 aactgactaa agtagtgagt tatacacagg gctgggatct attcttttta tcttttttta
     7741 ttctttcttt attctataaa ttataaccac ttgaatataa acaaaaaaaa cacacaaagg
     7801 tctagcggaa tttacagagg gtctagcaga atttacaagt tttccagcaa aggtctagca
     7861 gaatttacag atacccacaa ctcaaaggaa aaggactagt aattatcatt gactagccca
     7921 tctcaattgg tatagtgatt aaaattacct agaccaattg agatgtatgt ctgaattagt
     7981 tgttttcaaa gcaaatgaac tagcgattag tcgctatgac ttaacggagc atgaaaccaa
     8041 gctaatttta tgctgtgtgg cactactcaa ccccacgatt gaaaacccta caaggaaaga
     8101 acggacggta tcgttcactt ataaccaata cgctcagatg atgaacatca gtagggaaaa
     8161 tgcttatggt gtattagcta aagcaaccag agagctgatg acgagaactg tgaaaatcag
     8221 gaatcctttg gttaaaggct ttgagatttt ccagtggaca aactatgcca agttctcaag
     8281 cgaaaaatta gaattagttt ttagtgaaga gatattgcct tatcttttcc agttaaaaaa
     8341 attcataaaa tataatctgg aacatgttaa gtcttttgaa aacaaatact ctatgaggat
     8401 ttatgagtgg ttattaaaag aactaacaca aaagaaaact cacaaggcaa atatagagat
     8461 tagccttgat gaatttaagt tcatgttaat gcttgaaaat aactaccatg agtttaaaag
     8521 gcttaaccaa tgggttttga aaccaataag taaagattta aacacttaca gcaatatgaa
     8581 attggtggtt gataagcgag gccgcccgac tgatacgttg attttccaag ttgaactaga
     8641 tagacaaatg gatctcgtaa ccgaacttga gaacaaccag ataaaaatga atggtgacaa
     8701 aataccaaca accattacat cagattccta cctacgtaac ggactaagaa aaacactaca
     8761 cgatgcttta actgcaaaaa ttcagctcac cagttttgag gcaaaatttt tgagtgacat
     8821 gcaaagtaag catgatctca atggttcgtt ctcatggctc acgcaaaaac aacgaaccac
     8881 actagagaac atactggcta aatacggaag gatctga
//
