LOCUS       sequence-91618-e        8587 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector for FAK fret sensor without helix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8587)
  AUTHORS   Ritt M, Guan JL, Sivaramakrishnan S
  TITLE     Visualizing and manipulating focal adhesion kinase regulation in 
            live cells.
  JOURNAL   J Biol Chem. 2013 Mar 29;288(13):8875-86. doi: 
            10.1074/jbc.M112.421164. Epub 2013 Feb 7.
  PUBMED    23393139
REFERENCE   2  (bases 1 to 8587)
  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"; doi: 
            "10.1074/jbc.M112.421164"; journalName: "J Biol Chem"; date: 
            "2013-03-29- 29"; volume: "288"; issue: "13"; pages: "8875-86"
COMMENT     Sequence Label: pCS2-FAK0
FEATURES             Location/Qualifiers
     source          1..8587
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             37..60
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(79..81,82..792)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(79..81,82..792)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             2104..2124
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2143..2145,2146..2856)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4883..4903)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4883..4901)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4922..4938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4922..4938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4935..4957)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4946..4962
                     /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(4970..4976,4977..4994,4995..5000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5015..5036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5153..5170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5324..5912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5404..5423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6083..6874,6875..6943))
                     /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     6706..6725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6944..7048)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7074..7529)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7211..7232)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7423..7442
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7603..8587
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttatggact acaaggacga tgacgacaag
       61 ggaagcggag gaagcggagt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      121 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      181 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      241 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt cgcccgctac
      301 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      361 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      421 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      481 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
      541 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
      601 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
      661 ctgcccgaca accactacct gagctaccag tccaaactga gcaaagaccc caacgagaag
      721 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
      781 gagctgtaca agtctagagg aagcggagga agcggaggaa gcggaggaag cggaggaagc
      841 ggaatggcag cagcttacct tgatccaaac ttgaatcata caccaagttc aagtgcaaag
      901 acgcacctcg gtactgggat ggagcgttcc ccgggggcca tggagcgagt cctaaaggtt
      961 tttcactact ttgaaaacag cagcgagcca acgacgtggg ccagcattat ccggcatgga
     1021 gatgctactg atgttcgagg cataatacag aagattgtgg actgtcacaa agtgaaaaat
     1081 gtggcctgct atgggttgcg actcagtcat ctgcagtctg aggaggttca ctggctgcac
     1141 ctggacatgg gggtatccaa tgtgagagag aaatttgaac tagcacatcc tccagaagaa
     1201 tggaaatatg aactgagaat tcggtacctg cccaaaggat ttctaaacca gttcactgag
     1261 gacaaaccaa ctttaaattt tttctatcag caggtgaaaa atgactatat gttagaaata
     1321 gcagatcaag tggaccagga aattgctttg aaactaggtt gccttgaaat caggagatcc
     1381 tacggagaga tgagaggcaa tgcattagag aagaaatcca actatgaagt gctagaaaaa
     1441 gatgtcggtt taagacgatt ttttccgaag agtttgctag attcagtgaa ggccaaaaca
     1501 ctacgaaaat taatccaaca gacatttcga caatttgcca acctcaacag agaagaaagt
     1561 attttgaaat tctttgagat cctctctcca gtgtacagat ttgacaagga atgcttcaag
     1621 tgtgcccttg gttcaagctg gattatttca gtggagctgg caattggccc agaggaagga
     1681 atcagctacc ttacagacaa gggtgcaaat ccaactcacc tggcagattt taatcaagta
     1741 caaactattc agtattcaaa cagtgaagac aaggacagaa aagggatgtt gcaactgaag
     1801 atagctggtg cacctgagcc tctgacagtg acagcaccat ccttaaccat tgcagagaat
     1861 atggctgact tgatagacgg atactgccga ctggtgaatg gagccacgca atcttttatt
     1921 atcaggccac agaaagaagg tgaaagagct ttaccatcaa taccaaagct ggccaacaat
     1981 gagaagcaag gagtaaggtc gcacacagtc tctgtatcag aaacagatga ctatgcagag
     2041 ataatagatg aagaagatac ttatacaatg ggaagcggag gaagcggaga attcggaagc
     2101 ggagaaaacc tgtattttca gggcggaagc ggaggcgcgc ccgtgagcaa gggcgaggag
     2161 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2221 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     2281 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg
     2341 ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2401 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2461 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2521 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacgccatc
     2581 agcgacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     2641 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2701 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     2761 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2821 gccgggatca ctctcggcat ggacgagctg tacaagttaa ttaagggaag cggaggaagc
     2881 ggaggaagcg gaggaagcgg accatcaacc agagattatg aaattcaaag ggagagaatt
     2941 gaactggggc gctgcattgg tgaaggacag tttggagatg tgcaccaagg aatttacatg
     3001 agtccggaaa atccagctat ggctgtagca atcaaaacat gtaaaaactg cacctcagac
     3061 agcgttagag aaaagttcct acaagaagcc ttaacaatgc gtcagtttga tcatcctcac
     3121 attgtgaagc tcattggagt tattacagaa aacccagtgt ggataatcat ggagctctgt
     3181 acacttggag agttgagatc gtttctgcaa gtaagaaaat tcagcttgga cctggcctcc
     3241 ctcatcctct acgcttacca gcttagcaca gcacttgctt acctagagag caaaagattt
     3301 gtacatagag atattgctgc taggaacgtg ctggtatctg ccactgactg tgtgaaattg
     3361 ggtgactttg gcttatcccg atacatggaa gacagtactt actataaggc gtccaaagga
     3421 aagttaccta tcaaatggat ggctccagag tcaatcaact tccgacggtt tacctcagca
     3481 agcgatgtgt ggatgtttgg tgtgtgtatg tgggagatcc tgatgcatgg ggtaaagccc
     3541 ttccagggag tgaaaaataa tgatgttatt ggtcggattg agaacggtga gcggctcccc
     3601 atgcctccga actgccctcc caccctctac agccttatga ccaagtgctg ggcatacgac
     3661 cctagtagac gacccaggtt tactgaactt aaagcacaac tcagtacaat actggaggag
     3721 gagaagctgc agcaagagga acgaatgaga atggaatcca ggcgacaagt cacagtatcc
     3781 tgggactcag gaggatcaga tgaagctcct cccaagccca gcaggcctgg ttaccccagc
     3841 ccaaggtcca gtgaagggtt ttatccgagt cctcagcata tggtacagcc aaatcactac
     3901 caggtatctg gctactctgg ttctcatggg ataccagcca tggcaggcag catttatcct
     3961 gggcaggcgt ctctcttgga tcaaacagat tcctggaacc atcgacctca ggaagtatca
     4021 gcatggcagc caaacatgga ggattcgggc actttggatg tacgaggaat ggggcaggtt
     4081 ctgcccacac atctcatgga ggagaggtta ataagacaac agcaagagat ggaagaagat
     4141 caacgctggc ttgagaaaga ggaacgattc ctggtaatga aacctgatgt gcggctctcc
     4201 agaggcagca ttgaacggga ggacggaggt ctccagggcc cagctggtaa ccagcacata
     4261 tatcagcctg tgggtaaacc agatcatgcc gctccaccaa agaagccccc tcgccctgga
     4321 gccccccact tgggcagcct cgcgagcctg aacagccccg tggacagcta caacgaaggc
     4381 gtgaagatca agccacagga aatcagccct cctcctacgg ccaacctgga ccgctccaat
     4441 gacaaagtct atgagaatgt aaccgggctg gtgaaagctg tcatagagat gtccagtaaa
     4501 atacagccag ctccgccaga ggagtacgtg cccatggtaa aggaggttgg cttggcgctg
     4561 agaaccttgc tagcaacagt ggatgagtcg ctgccagtgc ttcctgcaag cacccacaga
     4621 gagattgaga tggcccagaa actgctgaac tctgacctgg ctgagctcat taacaagatg
     4681 aagctggccc agcagtacgt catgaccagc ctgcagcagg agtacaagaa gcaaatgctg
     4741 acggctgctc acgctctggc tgtggatgcc aagaacttgc tggatgtcat cgatcaagcc
     4801 agactgaaaa tgatcagcca gtccaggccc cactgagcgg ccgcggcgcc aatgcattgg
     4861 gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     4921 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4981 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5041 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5101 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5161 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5221 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5281 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5341 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5401 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5461 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5521 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5581 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5641 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5701 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5761 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5821 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5881 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5941 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6001 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6061 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6121 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6181 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6241 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6301 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6361 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6421 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6481 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6541 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6601 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6661 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6721 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6781 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6841 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6901 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6961 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7021 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     7081 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     7141 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     7201 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     7261 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     7321 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     7381 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7441 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7501 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     7561 ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccatag ccaattcaat
     7621 atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat
     7681 atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat
     7741 atggcggact tggcaccatg ccaattcaat atggcggact tggcactgtg ccaactgggg
     7801 aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg
     7861 ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa
     7921 tatataggca atatccaata tggccctatg ccaatatggc tattggccag gttcaatact
     7981 atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag
     8041 cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc
     8101 actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc
     8161 ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc
     8221 tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc
     8281 attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca
     8341 cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact attggaagga
     8401 cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg cccgcgatgg
     8461 ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc
     8521 attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca atgctcgttt
     8581 agggaac
//
