LOCUS       sequence-365925-        7915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expressing FAK in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7915)
  AUTHORS   Case LB, De Pasquale M, Henry L, Rosen MK
  TITLE     Synergistic phase separation of two pathways promotes integrin 
            clustering and nascent adhesion formation.
  JOURNAL   Elife. 2022 Jan 20;11. pii: 72588. doi: 10.7554/eLife.72588.
  PUBMED    35049497
REFERENCE   2  (bases 1 to 7915)
  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"; journalName: "Elife."; 
            date: "2022-01-20"; volume: "11. pii: 72588. doi"; pages: " 
            10.7554/eLife.72588"
COMMENT     Sequence Label: GFP-FAK W266A
FEATURES             Location/Qualifiers
     source          1..7915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1310..1319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1316..1318,1319..1321,1322..2032)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1361..1382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1622..1641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1969..1990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5406..5527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5443..5462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5497..5516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5534..5989)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5671..5692)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5883..5902
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6016..6120
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6122..6479
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6122..6313
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6145..6165)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6330..6465
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6514..7308
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6514..7308
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6568..6587)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7178..7197
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7496..7515)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7540..7587
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     1321 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1381 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1441 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1501 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1561 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1621 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1681 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1741 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1801 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1861 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1921 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1981 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact
     2041 cagatcgggc ggttccggga gcggaatggc agctgcttat cttgacccaa acttgaatca
     2101 cacaccaagt tcgagtacta agactcacct gggtactggc atggaacggt cccctggtgc
     2161 aatggaacga gtattaaagg tctttcatca ttttgaaagc agtagtgagc caaccacctg
     2221 ggccagtatt atcaggcatg gagatgctac tgatgtcagg ggcatcattc agaagatagt
     2281 ggacagtcac aaagtaaagc acgtggcctg ctatggattt cgcctcagtc acctgcgatc
     2341 ggaggaggtg cactggctgc acgtggacat gggtgtctcc agtgtgaggg agaaatatga
     2401 actcgcccac ccaccagagg agtggaaata tgagttgaga attcgttact tgccaaaagg
     2461 atttctaaac cagtttactg aagataagcc aacattgaat ttcttctatc aacaggtgaa
     2521 gagtgactac atgcaagaaa tagctgatca agtagaccaa gaaatagctt tgaagttggg
     2581 ttgtttggaa attaggcgat cctattggga gatgaggggt aatgccctag agaagaagtc
     2641 caactatgaa gtattagaaa aagatgttgg tttaaagcga ttttttccta agagtttact
     2701 ggattctgtc aaggccaaaa cactaagaaa gctgatccag caaacattca ggcagtttgc
     2761 caaccttaat agagaagaaa gcatcctgaa attctttgag attctttctc ctgtgtacag
     2821 atttgacaaa gagtgcttca agtgtgccct tgggtcaagt gcgatcattt ctgtggaatt
     2881 ggcaatcggc ccagaagaag ggatcagtta cctgacagac aaaggctgca atcccacaca
     2941 cctagcagac tttaaccaag tgcagaccat ccagtactca aacagtgaag acaaagacag
     3001 gaaaggaatg ctacaactca aaatcgctgg tgctcctgag cctctgaccg tgacggcacc
     3061 atccctaacc attgccgaga acatggccga cctgattgat ggatactgcc ggctggtgaa
     3121 tggagccaca caatctttca tcatcagacc ccagaaagaa ggtgaacggg cgttgccatc
     3181 aataccaaag ttggccaaca gtgaaaagca aggcatgcgg acacatgcag tctctgtgtc
     3241 agagacagat gactatgcag agatcatcga tgaggaagac acatacacca tgccctcgac
     3301 cagggattat gagattcaga gagaaagaat agaacttgga cgctgtattg gagaaggtca
     3361 gtttggagat gtacatcaag gcgtgtacct gagcccagag aatccagctt tggctgttgc
     3421 aatcaaaaca tgtaaaaact gtacttcgga cagcgtgaga gagaagttcc ttcaagaagc
     3481 cttaacaatg cgtcagtttg accatcctca catcgtgaag ctgattggag tcattacaga
     3541 gaaccctgtc tggataatca tggagttgtg cacacttgga gagctgaggt catttttgca
     3601 agtaagaaaa tacagcttgg acctggcatc tttgatatta tatgcctatc agcttagtac
     3661 agcactcgcg tatctggaga gcaaaagatt tgttcacagg gacattgctg ctcggaatgt
     3721 tctggtgtct tcaaatgatt gtgtaaaatt gggagacttt ggattatctc gatatatgga
     3781 agacagtact tactataaag cttccaaagg aaaattacct attaaatgga tggctccaga
     3841 gtcaatcaat tttcgacgtt ttacctcagc tagtgacgtg tggatgtttg gtgtgtgtat
     3901 gtgggagata ctgatgcatg gcgtgaagcc ttttcaagga gtgaagaaca atgatgtgat
     3961 cggtcgaatt gaaaatgggg aaagattacc aatgcctcca aattgtcctc ccaccctcta
     4021 cagccttatg acgaaatgtt gggcctatga ccccagcagg cggcccaggt ttactgaact
     4081 aaaagctcag ctcagcacaa tcctggagga ggagaaggtg cagcaagaag aacggatgag
     4141 gatggaatcc agaagacagg ctaccgtgtc ctgggactct ggagggtctg atgaagcacc
     4201 acccaagccc agcagacctg gttatcctag cccgagatcc agtgaaggat tttatcccag
     4261 cccacagcac atggtccaga ccaatcacta ccaggtctct ggctacccgg gttcccatgg
     4321 aatcccagcc atggctggca gcatttacca aggtcaggca tctcttctgg accagacaga
     4381 attgtggaat cacagacctc aggagatgtc catgtggcag cccagtgtgg aggattcggc
     4441 agctttggac cttcgaggga tgggacaggt gcttccccct cacctgatgg aagagcggct
     4501 aatccgacag cagcaggaaa tggaagaaga ccagcgctgg ctggagaagg aagagcgatt
     4561 cctgaaacct gatgtgaggc tctctcgagg cagcatcgac agggaagacg ggagttttca
     4621 gggtccgact ggaaaccaac acatctatca gcctgtgggg aagccagatc ctgcagctcc
     4681 accaaagaaa cctcctcgcc ctggagcacc tggccaccta agcaacctgt ccagcatcag
     4741 cagccctgca gacagctaca atgagggtgt caagcttcag ccccaggaaa tcagcccccc
     4801 tcccactgcc aaccttgacc ggtccaatga caaggtatat gaaaatgtga caggcctagt
     4861 gaaggctgtc atcgagatgt ccagcaaaat ccagccagct cctccagaag agtacgtccc
     4921 tatggtgaag gaagttggcc tggccctgcg gaccttacta gccacggtgg atgagaccat
     4981 tcctgctctt ccagccagca ctcatcgaga gatcgagatg gcacagaagc tgctgaactc
     5041 cgacttaggc gagctcatca gcaagatgaa gctggcgcag cagtacgtca tgaccagcct
     5101 gcagcaggag tataagaagc agatgctgac agctgctcac gccctggctg tggatgccaa
     5161 gaacctactt gatgttattg atcaagcaag actgaaaatg ctagggcaga cacggccaca
     5221 ctgataatag tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
     5281 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     5341 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     5401 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5461 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5521 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5581 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5641 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5701 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5761 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5821 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5881 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5941 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     6001 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6061 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6121 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     6181 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     6241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6301 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6361 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6421 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     6481 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     6541 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6601 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6661 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     6721 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6781 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6841 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6901 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6961 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     7021 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     7081 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     7141 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     7201 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     7261 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     7321 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     7381 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     7441 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     7501 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     7561 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     7621 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     7681 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     7741 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     7801 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7861 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
