LOCUS       sequence-475725-        6388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, Excitation: 587, Emission: 610.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6388)
  AUTHORS   Paszek MJ, DuFort CC, Rubashkin MG, Davidson MW, Thorn KS, Liphardt 
            JT, Weaver VM
  TITLE     Scanning angle interference microscopy reveals cell dynamics at the 
            nanoscale.
  JOURNAL   Nat Methods. 2012 Jul 1;9(8):825-7. doi: 10.1038/nmeth.2077.
  PUBMED    22751201
REFERENCE   2  (bases 1 to 6388)
  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: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-07-1- 1"; volume: "9"; 
            issue: "8"; pages: "825-7"
COMMENT     Sequence Label: mCherry-Paxillin-22
FEATURES             Location/Qualifiers
     source          1..6388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2217..2269
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2277..2286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2283..2993
                     /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(2428..2446)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2696..2715
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3116..3237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3153..3172)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3207..3226
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3244..3699)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3381..3402)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3593..3612
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3726..3830
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3832..4189
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3832..4023
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3855..3875)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4040..4175
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4224..5018
                     /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             4224..5018
                     /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(4278..4297)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4888..4907
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5206..5225)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5250..5297
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5626..6214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6115..6134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      541 tggacgacct cgatgcctta ctggcagacc tggaatccac cacctcgcat atctctaagc
      601 gaccggtgtt tctaacagag gaaacgcctt actcctaccc aactggaaac catacgtacc
      661 aggagattgc tgtgccaccg ccggtgcccc cgccgccttc cagtgaggcc ctaaatggca
      721 ctgtgattga ccccttagac cagtggcaac ccagcgtgtc cagatacggc caccagcaac
      781 ctcagtccca gtctccgata tacagctcta gtgccaaaag ctccagtgcc tctgttccta
      841 gagacgggct cagctcccct tctccccgtg ccagtgagga ggaacacgtg tacagcttcc
      901 caaacaagca gaagtctgca gaaccatctc ccacaatgac cagcacctcc ctgggcagca
      961 acctctcgga actggacagg cttctcctgg aactgaatgc tgttcaacat aatcccccca
     1021 gtggcttctc agcagatgaa gtcagcagaa gcccatcact gcccaatgtg accggacctc
     1081 actatgtcat cccagagagc agcagctctg caggagggaa ggctgcaccc cctacaaaag
     1141 aaaaaccaaa gcggaatggt gggcgtggga ttgaagatgt gcgtcccagt gtggagagcc
     1201 tgctggatga gctggagagc tctgtgccaa gcccagtccc tgcaatcact gtgagccaag
     1261 gggaggtgag cagcccccag cgtgtcaacg ccagtcagca gcagacccgt atctctgctt
     1321 cttcagctac acgagaactg gatgagctga tggcgtccct ctctgacttt aagttcatgg
     1381 cacaggggaa agccggcggc agcagctccc caccatccac caccccaaag cctggcagtc
     1441 agctggacac catgctggga agtctgcagt ctgacctgaa caaactgggg gtagcgacgg
     1501 tcgccaaggg tgtgtgcgga gcctgcaaga agccgattgc tgggcaggta gttacagcca
     1561 tggggaagac ctggcaccct gagcacttcg tctgcaccca ctgccaggag gagattggat
     1621 cacggaactt ctttgagcgg gatgggcagc cctactgtga gaaggactat cacaacctct
     1681 tctctcctcg ctgctactac tgcaatgggc caatccttga taaagtggtg acggctttgg
     1741 acaggacgtg gcaccctgaa cactttttct gtgcccagtg tggagttttc tttggacctg
     1801 aaggatttca tgagaaggat ggcaaagcct attgccgcaa ggactacttt gacatgtttg
     1861 ctcccaagtg tggaggctgt gcccgggcta tcctggaaaa ttacatttct gccttaaaca
     1921 ccctgtggca ccctgagtgc tttgtctgtc gggaatgttt cacaccgttc atcaacggca
     1981 gcttcttcga gcacgacggg cagccctact gcgaggtgca ttaccacgag cgccgcggct
     2041 cgctgtgctc tggctgccag aagcccatca cgggacgctg catcactgcc atgggcaaga
     2101 aatttcaccc cgaacacttt gtctgtgcct tctgcctcaa gcagctcaac aaaggaacct
     2161 tcaaagaaca gaacgacaag ccctactgtc agaactgctt tctcaaactc ttctgtctcg
     2221 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     2281 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     2341 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     2401 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     2461 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     2521 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     2581 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2641 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2701 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2761 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2821 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2881 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2941 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taaagcggcc
     3001 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3061 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3241 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3301 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3361 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3421 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3481 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3541 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3601 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3661 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3721 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3781 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3841 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3901 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3961 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4021 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4081 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4141 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4201 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4261 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4321 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4381 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4441 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4501 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4561 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4621 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4681 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4741 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4801 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4861 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4921 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4981 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5041 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5101 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5161 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5221 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5281 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5341 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5401 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5461 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5521 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5581 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5641 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5701 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5761 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5821 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5881 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5941 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6001 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6061 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6121 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6181 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6241 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6301 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
     6361 tagttcatag cccatatatg gagttccg
//
