LOCUS       sequence-91620-e        9004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector for FAK fret sensor with 20 nm helix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9004)
  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 9004)
  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-FAK20
FEATURES             Location/Qualifiers
     source          1..9004
                     /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             2113..2133
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2560..2562,2563..3273)
                     /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(5300..5320)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5300..5318)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5339..5355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5339..5355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5352..5374)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5363..5379
                     /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(5387..5393,5394..5411,5412..5417))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5432..5453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5570..5587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5741..6329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5821..5840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6500..7291,7292..7360))
                     /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     7123..7142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7361..7465)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7491..7946)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7628..7649)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7840..7859
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        8020..9004
                     /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 ggaggaagcg gagaaaacct gtattttcag ggcggaagcg gagaagagga agagaagaag
     2161 aaagaagagg aagaaaagaa acaaaaagaa gaacaagaaa gacttgcaaa agaagaggca
     2221 gagagaaaac aaaaagaaga acaagaaaga cttgaaagag aaagaaaaga gagagaagaa
     2281 caagaaaaga aagccaaaga agaggcagag agaattgcaa agttagaggc tgaaaagaag
     2341 gcagaagaag aaagaaaagc caaagaagaa gaagagagaa aagccaaaga agaagaggaa
     2401 agaaagaaga aagaggagca agaaagactt gcaaaagaaa aggaagaagc agaaagaaaa
     2461 gctgcagagg aaaagaaagc taaagaagaa caagaaagaa aagaaaagga agaagcagaa
     2521 agaaaacaaa gaggctctgg cggctctggc ggcgcgcccg tgagcaaggg cgaggagctg
     2581 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2641 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2701 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctggggc
     2761 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2821 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2881 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2941 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa cgccatcagc
     3001 gacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
     3061 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3121 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     3181 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3241 gggatcactc tcggcatgga cgagctgtac aagttaatta agggaagcgg aggaagcgga
     3301 ggaagcggag gaagcggacc atcaaccaga gattatgaaa ttcaaaggga gagaattgaa
     3361 ctggggcgct gcattggtga aggacagttt ggagatgtgc accaaggaat ttacatgagt
     3421 ccggaaaatc cagctatggc tgtagcaatc aaaacatgta aaaactgcac ctcagacagc
     3481 gttagagaaa agttcctaca agaagcctta acaatgcgtc agtttgatca tcctcacatt
     3541 gtgaagctca ttggagttat tacagaaaac ccagtgtgga taatcatgga gctctgtaca
     3601 cttggagagt tgagatcgtt tctgcaagta agaaaattca gcttggacct ggcctccctc
     3661 atcctctacg cttaccagct tagcacagca cttgcttacc tagagagcaa aagatttgta
     3721 catagagata ttgctgctag gaacgtgctg gtatctgcca ctgactgtgt gaaattgggt
     3781 gactttggct tatcccgata catggaagac agtacttact ataaggcgtc caaaggaaag
     3841 ttacctatca aatggatggc tccagagtca atcaacttcc gacggtttac ctcagcaagc
     3901 gatgtgtgga tgtttggtgt gtgtatgtgg gagatcctga tgcatggggt aaagcccttc
     3961 cagggagtga aaaataatga tgttattggt cggattgaga acggtgagcg gctccccatg
     4021 cctccgaact gccctcccac cctctacagc cttatgacca agtgctgggc atacgaccct
     4081 agtagacgac ccaggtttac tgaacttaaa gcacaactca gtacaatact ggaggaggag
     4141 aagctgcagc aagaggaacg aatgagaatg gaatccaggc gacaagtcac agtatcctgg
     4201 gactcaggag gatcagatga agctcctccc aagcccagca ggcctggtta ccccagccca
     4261 aggtccagtg aagggtttta tccgagtcct cagcatatgg tacagccaaa tcactaccag
     4321 gtatctggct actctggttc tcatgggata ccagccatgg caggcagcat ttatcctggg
     4381 caggcgtctc tcttggatca aacagattcc tggaaccatc gacctcagga agtatcagca
     4441 tggcagccaa acatggagga ttcgggcact ttggatgtac gaggaatggg gcaggttctg
     4501 cccacacatc tcatggagga gaggttaata agacaacagc aagagatgga agaagatcaa
     4561 cgctggcttg agaaagagga acgattcctg gtaatgaaac ctgatgtgcg gctctccaga
     4621 ggcagcattg aacgggagga cggaggtctc cagggcccag ctggtaacca gcacatatat
     4681 cagcctgtgg gtaaaccaga tcatgccgct ccaccaaaga agccccctcg ccctggagcc
     4741 ccccacttgg gcagcctcgc gagcctgaac agccccgtgg acagctacaa cgaaggcgtg
     4801 aagatcaagc cacaggaaat cagccctcct cctacggcca acctggaccg ctccaatgac
     4861 aaagtctatg agaatgtaac cgggctggtg aaagctgtca tagagatgtc cagtaaaata
     4921 cagccagctc cgccagagga gtacgtgccc atggtaaagg aggttggctt ggcgctgaga
     4981 accttgctag caacagtgga tgagtcgctg ccagtgcttc ctgcaagcac ccacagagag
     5041 attgagatgg cccagaaact gctgaactct gacctggctg agctcattaa caagatgaag
     5101 ctggcccagc agtacgtcat gaccagcctg cagcaggagt acaagaagca aatgctgacg
     5161 gctgctcacg ctctggctgt ggatgccaag aacttgctgg atgtcatcga tcaagccaga
     5221 ctgaaaatga tcagccagtc caggccccac tgagcggccg cggcgccaat gcattgggcc
     5281 cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     5341 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5401 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5461 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5521 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5581 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5641 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5701 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5761 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5821 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5881 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5941 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6001 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6061 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6121 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6181 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6241 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6301 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6361 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6421 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6481 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6541 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6601 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6661 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6721 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6781 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6841 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6901 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6961 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7021 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7081 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7141 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7201 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7261 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7321 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7381 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7441 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     7501 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     7561 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     7621 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     7681 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     7741 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     7801 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     7861 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     7921 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     7981 agggcgatcg gtgcgggcct cttcgctatt acgccagtcg accatagcca attcaatatg
     8041 gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg
     8101 gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg
     8161 gcggacttgg caccatgcca attcaatatg gcggacttgg cactgtgcca actggggagg
     8221 ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca
     8281 tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat
     8341 ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg
     8401 tattggccct atgccatata gtattccata tatgggtttt cctattgacg tagatagccc
     8461 ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc catagccact
     8521 cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt
     8581 cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg
     8641 ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt
     8701 gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt
     8761 ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt ggaaggacgc
     8821 cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg
     8881 ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt
     8941 gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg
     9001 gaac
//
