LOCUS       sequence-328409-        6712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Focal Adhesions, Excitation: 448 / 473, Emission: 500 
            / 528.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6712)
  AUTHORS   Shu X, Lev-Ram V, Deerinck TJ, Qi Y, Ramko EB, Davidson MW, Jin Y, 
            Ellisman MH, Tsien RY
  TITLE     A genetically encoded tag for correlated light and electron 
            microscopy of intact cells, tissues, and organisms.
  JOURNAL   PLoS Biol. 2011 Apr;9(4):e1001041. doi: 
            10.1371/journal.pbio.1001041. Epub 2011 Apr 5.
  PUBMED    21483721
REFERENCE   2  (bases 1 to 6712)
  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: "PLoS 
            Biol."; date: "2011-04"; volume: "9(4):e1001041. doi"; pages: " 
            10.1371/journal.pbio.1001041. Epub 2011 Apr 5"
COMMENT     Sequence Label: mCherry-miniSOG-Paxillin-22
FEATURES             Location/Qualifiers
     source          1..6712
                     /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..2990
                     /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"
     CDS             2997..3317
                     /codon_start=1
                     /product="mini singlet oxygen generator, a fluorescent 
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
                     /label=miniSOG
                     /translation="MEKSFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNGRFLQG
                     PETDQATVQKIRDAIRDQREITVQLINYTKSGKKFWNLLHLQPMRDQKGELQYFIGVQL
                     DG"
     polyA_signal    3440..3561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3477..3496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3531..3550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3568..4023)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3705..3726)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3917..3936
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4050..4154
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4156..4513
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4156..4347
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4179..4199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4364..4499
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4548..5342
                     /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             4548..5342
                     /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(4602..4621)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5212..5231
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5530..5549)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5574..5621
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5950..6538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6439..6458
                     /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 ggcggcatgg
     3001 agaagagctt cgtgatcacc gacccccgcc tgcccgacaa ccccatcatc ttcgccagcg
     3061 acggcttcct ggagctgacc gagtacagca gagaggagat cctgggccgc aacggccgct
     3121 tcctgcaggg ccccgagacc gaccaggcca ccgtgcagaa gatccgcgac gccatccgcg
     3181 accagcgcga gatcaccgtg cagctgatca actacaccaa gagcggcaag aagttctgga
     3241 acctgctgca cctgcagccc atgcgcgacc agaagggcga gctgcagtac ttcatcggcg
     3301 tgcagctgga cggctaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     3361 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3421 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3481 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3541 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3601 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3661 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3721 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3781 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3841 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3901 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3961 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4021 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4081 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4141 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4201 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4261 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4321 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4381 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4441 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4501 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4561 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4621 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4681 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4741 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4801 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4861 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4921 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4981 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5041 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5101 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5161 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5221 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5281 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5341 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5401 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5461 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5521 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5581 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5641 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5701 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5761 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5821 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5881 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5941 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6001 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6061 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6121 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6181 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6241 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6301 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6361 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6421 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6481 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6541 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6601 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     6661 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cg
//
