LOCUS       sequence-42587-e        7913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7913)
  AUTHORS   Chan KT, Bennin DA, Huttenlocher A
  TITLE     Regulation of adhesion dynamics by calpain-mediated proteolysis of 
            focal adhesion kinase (FAK).
  JOURNAL   J Biol Chem. 2010 Apr 9;285(15):11418-26. Epub 2010 Feb 11.
  PUBMED    20150423
REFERENCE   2  (bases 1 to 7913)
  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: "J Biol 
            Chem."; date: "2010-04-9"; volume: "285(15)"; pages: "11418-26. Epub
            2010 Feb 11"
COMMENT     Sequence Label: pmCherry-C1-FAK-HA-V744G
FEATURES             Location/Qualifiers
     source          1..7913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             603..1310
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(748..766)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1016..1035
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4479..4505
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4509..4535
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4701..4822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4738..4757)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4792..4811
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4829..5284)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4966..4987)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5178..5197
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5311..5415
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5417..5774
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5417..5608
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5440..5460)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5625..5760
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5809..6603
                     /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             5809..6603
                     /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(5863..5882)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6473..6492
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6791..6810)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6835..6882
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7211..7799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7700..7719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
      661 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
      721 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
      781 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
      841 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
      901 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
      961 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1021 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     1081 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     1141 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     1201 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1261 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag agatctatgg
     1321 cagctgctta tcttgaccca aacttgaatc acacaccaag ttcgagtact aagactcacc
     1381 tgggtactgg catggaacgg tcccctggtg caatggaacg agtattaaag gtctttcatc
     1441 attttgaaag cagtagtgag ccaaccacct gggccagtat tatcaggcat ggagatgcta
     1501 ctgatgtcag gggcatcatt cagaagatag tggacagtca caaagtaaag cacgtggcct
     1561 gctatggatt tcgcctcagt cacctgcgat cggaggaggt gcactggctg cacgtggaca
     1621 tgggtgtctc cagtgtgagg gagaaatatg aactcgccca cccaccagag gagtggaaat
     1681 atgagttgag aattcgttac ttgccaaaag gatttctaaa ccagtttact gaagataagc
     1741 caacattgaa tttcttctat caacaggtga agagtgacta catgcaagaa atagctgatc
     1801 aagtagacca agaaatagct ttgaagttgg gttgtttgga aattaggcga tcctattggg
     1861 agatgagggg taatgcccta gagaagaagt ccaactatga agtattagaa aaagatgttg
     1921 gtttaaagcg attttttcct aagagtttac tggattctgt caaggccaaa acactaagaa
     1981 agctgatcca gcaaacattc aggcagtttg ccaaccttaa tagagaagaa agcatcctga
     2041 aattctttga gattctttct cctgtgtaca gatttgacaa agagtgcttc aagtgtgccc
     2101 ttgggtcaag ttggatcatt tctgtggaat tggcaatcgg cccagaagaa gggatcagtt
     2161 acctgacaga caaaggctgc aatcccacac acctagcaga ctttaaccaa gtgcagacca
     2221 tccagtactc aaacagtgaa gacaaagaca ggaaaggaat gctacaactc aaaatcgctg
     2281 gtgctcctga gcctctgacc gtgacggcac catccctaac cattgccgag aacatggccg
     2341 acctgattga tggatactgc cggctggtga atggagccac acaatctttc atcatcagac
     2401 cccagaaaga aggtgaacgg gcgttgccat caataccaaa gttggccaac agtgaaaagc
     2461 aaggcatgcg gacacatgca gtctctgtgt cagagacaga tgactatgca gagatcatcg
     2521 atgaggaaga cacatacacc atgccctcga ccagggatta tgagattcag agagaaagaa
     2581 tagaacttgg acgctgtatt ggagaaggtc agtttggaga tgtacatcaa ggcgtgtacc
     2641 tgagcccaga gaatccagct ttggctgttg caatcaaaac atgtaaaaac tgtacttcgg
     2701 acagcgtgag agagaagttc cttcaagaag ccttaacaat gcgtcagttt gaccatcctc
     2761 acatcgtgaa gctgattgga gtcattacag agaaccctgt ctggataatc atggagttgt
     2821 gcacacttgg agagctgagg tcatttttgc aagtaagaaa atacagcttg gacctggcat
     2881 ctttgatatt atatgcctat cagcttagta cagcactcgc gtatctggag agcaaaagat
     2941 ttgttcacag ggacattgct gctcggaatg ttctggtgtc ttcaaatgat tgtgtaaaat
     3001 tgggagactt tggattatct cgatatatgg aagacagtac ttactataaa gcttccaaag
     3061 gaaaattacc tattaaatgg atggctccag agtcaatcaa ttttcgacgt tttacctcag
     3121 ctagtgacgt gtggatgttt ggtgtgtgta tgtgggagat actgatgcat ggcgtgaagc
     3181 cttttcaagg agtgaagaac aatgatgtga tcggtcgaat tgaaaatggg gaaagattac
     3241 caatgcctcc aaattgtcct cccaccctct acagccttat gacgaaatgt tgggcctatg
     3301 accccagcag gcggcccagg tttactgaac taaaagctca gctcagcaca atcctggagg
     3361 aggagaaggt gcagcaagaa gaacggatga ggatggaatc cagaagacag gctaccgtgt
     3421 cctgggactc tggagggtct gatgaagcac cacccaagcc cagcagacct ggttatccta
     3481 gcccgagatc cagtgaagga ttttatccca gcccacagca catggtccag accaatcact
     3541 accagggctc tggctacccg ggttcccatg gaatcccagc catggctggc agcatttacc
     3601 aaggtcaggc atctcttctg gaccagacag aattgtggaa tcacagacct caggagatgt
     3661 ccatgtggca gcccagtgtg gaggattcgg cagctttgga ccttcgaggg atgggacagg
     3721 tgcttccccc tcacctgatg gaagagcggc taatccgaca gcagcaggaa atggaagaag
     3781 accagcgctg gctggagaag gaagagcgat tcctgaaacc tgatgtgagg ctctctcgag
     3841 gcagcatcga cagggaagac gggagttttc agggtccgac tggaaaccaa cacatctatc
     3901 agcctgtggg gaagccagat cctgcagctc caccaaagaa acctcctcgc cctggagcac
     3961 ctggccacct aagcaacctg tccagcatca gcagccctgc agacagctac aatgagggtg
     4021 tcaagcttca gccccaggaa atcagccccc ctcccactgc caaccttgac cggtccaatg
     4081 acaaggtata tgagaatgtg acaggcctag tgaaggctgt catcgagatg tccagcaaaa
     4141 tccagccagc tcctccagaa gagtacgtcc ctatggtgaa ggaagttggc ctggccctgc
     4201 ggaccttact agccacggtg gatgagacca ttcctgctct tccagccagc actcatcgag
     4261 agatcgagat ggcacagaag ctgctgaact ccgacttagg cgagctcatc agcaagatga
     4321 agctggcgca gcagtacgtc atgaccagcc tgcagcagga gtataagaag cagatgctga
     4381 cagctgctca cgccctggct gtggatgcca agaacctact tgatgttatt gatcaagcaa
     4441 gactgaaaat gctagggcag acacgcggcc gcatctttta cccatacgat gttcctgact
     4501 atgcgggcta tccctatgac gtcccggact atgcaggatc caccggatct agataaggta
     4561 ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat accacatttg
     4621 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     4681 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     4741 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4801 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
     4861 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4921 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4981 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5041 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     5101 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5161 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5221 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5281 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5341 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5401 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     5461 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5521 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5581 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5641 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5701 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5761 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5821 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5881 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5941 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     6001 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6061 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6121 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6181 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6241 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6301 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     6361 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6421 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6481 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6541 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6601 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6661 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6721 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     6781 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     6841 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     6901 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     6961 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     7021 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     7081 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7141 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7201 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7261 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7321 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     7381 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7441 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7501 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7561 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7621 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7681 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7741 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7801 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7861 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
