LOCUS       sequence-201677-        6738 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 6738)
  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 6738)
  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: PA-mCherry-miniSOG-Zyxin-6
FEATURES             Location/Qualifiers
     source          1..6738
                     /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"
     regulatory      574..583
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2304..2313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2310..3017
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein (Subach et al., 2009)"
                     /label=PAmCherry1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2455..2473)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             3024..3344
                     /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    3466..3587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3503..3522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3557..3576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3594..4049)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3731..3752)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3943..3962
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4076..4180
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4182..4539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4182..4373
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4205..4225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4390..4525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4574..5368
                     /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             4574..5368
                     /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(4628..4647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5238..5257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5556..5575)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5600..5647
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5976..6564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6465..6484
                     /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 gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attccatggc ggccccccgc ccgtctcccg
      601 cgatccccgt ttcggtctcg gctccggctt tttacgcccc gcagaagaag ttcggccctg
      661 tggtggcccc aaagcccaaa gtgaatccct tccggcccag ggacagcgag cctcccccgg
      721 cacccggggc ccagcgcgca cagatgggcc gggtgggcga tattcccccg ccgcccccgg
      781 aagactttcc cctgcctcca cctccccttg ctggggatgg cgacgatgca gagggtgctc
      841 tgggaggtgc cttcccgccg ccccctcccc cgatcgagga atcatttccc cctgcgcctc
      901 tggaggagga gatcttccct tccccgccgc ctcctccgga ggaggaggga gggcctgagg
      961 cccccatacc gcccccacca cagcccaggg agaaggtgag cagtattgat ttggagatcg
     1021 actctctgtc ctcactgctg gatgacatga ccaagaatga tcctttcaaa gcccgggtgt
     1081 catctggata tgtgccccca ccagtggcca ctccattcag ttccaagtcc agtaccaagc
     1141 ctgcagccgg gggcacagca cccctgcctc cttggaagtc cccttccagc tcccagcctc
     1201 tgccccaggt tccggctccg gctcagagcc agacacagtt ccatgttcag ccccagcccc
     1261 agcccaagcc tcaggtccaa ctccatgtcc agtcccagac ccagcctgtg tctttggcta
     1321 acacccagcc ccgagggccc ccagcctcat ctccggctcc agcccctaag ttttctccag
     1381 tgactcctaa gtttactcct gtggcttcca agttcagtcc tggagcccca ggtggatctg
     1441 ggtcacaacc aaatcaaaaa ttggggcacc ccgaagctct ttctgctggc acaggctccc
     1501 ctcaacctcc cagcttcacc tatgcccagc agagggagaa gccccgagtg caggagaagc
     1561 agcaccccgt gcccccaccg gctcagaacc aaaaccaggt gcgctcccct ggggccccag
     1621 ggcccctgac tctgaaggag gtggaggagc tggagcagct gacccagcag ctaatgcagg
     1681 acatggagca tcctcagagg cagaatgtgg ctgtcaacga actctgcggc cgatgccatc
     1741 aacccctggc ccgggcgcag ccagccgtcc gcgctctagg gcagctgttc cacatcgcct
     1801 gcttcacctg ccaccagtgt gcgcagcagc tccagggcca gcagttctac agtctggagg
     1861 gggcgccgta ctgcgagggc tgttacactg acaccctgga gaagtgtaac acctgcgggg
     1921 agcccatcac tgaccgcatg ctgagggcca cgggcaaggc ctatcacccg cactgcttca
     1981 cctgtgtggt ctgcgcccgc cccctggagg gcacctcctt catcgtggac caggccaacc
     2041 ggccccactg tgtccccgac taccacaagc agtacgcccc gaggtgctcc gtctgctctg
     2101 agcccatcat gcctgagcct ggccgagatg agactgtgcg agtggtcgcc ctggacaaga
     2161 acttccacat gaagtgttac aagtgtgagg actgcgggaa gcccctgtcg attgaggcag
     2221 atgacaatgg ctgcttcccc ctggacggtc acgtgctctg tcggaagtgc cacactgcta
     2281 gagcccagac ggatccaccg gtcgccacca tggtgagcaa gggcgaggag gataacatgg
     2341 ccatcattaa ggagttcatg cgcttcaagg tgcacatgga ggggtccgtg aacggccacg
     2401 tgttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     2461 tgaaggtgac caagggcggc cccctgccct tcacctggga catcctgagc cctcagttca
     2521 tgtacggctc caatgcctac gtgaagcacc ccgccgacat ccccgactac tttaagctgt
     2581 ccttccccga gggcttcaag tgggagcgcg tgatgaaatt cgaggacggc ggcgtggtga
     2641 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     2701 gcaccaactt cccctccgac ggccccgtga tgcagaagaa gaccatgggc tgggaggccc
     2761 tctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgaggtcaag cccagagtga
     2821 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     2881 ccgtgcagct gcccggcgcc tacaacgtca accgcaagct ggacatcacc agccacaacg
     2941 aggactacac catcgtggag cagtacgaga gagccgaggg ccgccactcc accggcggca
     3001 tggacgagct gtacaagggc ggcatggaga agagcttcgt gatcaccgac ccccgcctgc
     3061 ccgacaaccc catcatcttc gccagcgacg gcttcctgga gctgaccgag tacagcagag
     3121 aggagatcct gggccgcaac ggccgcttcc tgcagggccc cgagaccgac caggccaccg
     3181 tgcagaagat ccgcgacgcc atccgcgacc agcgcgagat caccgtgcag ctgatcaact
     3241 acaccaagag cggcaagaag ttctggaacc tgctgcacct gcagcccatg cgcgaccaga
     3301 agggcgagct gcagtacttc atcggcgtgc agctggacgg ctaagcggcc gcgactctag
     3361 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3421 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3481 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3541 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     3601 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3661 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3721 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3781 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3841 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3901 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3961 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4021 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4081 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4141 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4201 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4261 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4381 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4441 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4501 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4561 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4621 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4681 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4741 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4801 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4861 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4921 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4981 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5041 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5101 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5161 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5221 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5281 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5341 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5401 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5461 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5521 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5581 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5641 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5701 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5761 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5821 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5881 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5941 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6001 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6061 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6121 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6661 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     6721 cccatatatg gagttccg
//
