LOCUS       sequence-253128-        6359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Gap Junctions, Excitation: 448 / 473, Emission: 500 / 
            528.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6359)
  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 6359)
  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-Cx43-7
FEATURES             Location/Qualifiers
     source          1..6359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        89..392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        393..596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     547..567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2013..2022
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2019..2726
                     /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(2164..2182)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             2733..3053
                     /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    3175..3296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3212..3231)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3266..3285
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3303..3758)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3440..3461)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3652..3671
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3785..3889
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3891..4248
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3891..4082
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3914..3934)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4099..4234
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4283..5077
                     /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             4283..5077
                     /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(4337..4356)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4947..4966
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5265..5284)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5309..5356
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5685..6273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6174..6193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
       61 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      121 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      181 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      241 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      301 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      361 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      421 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      481 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      541 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
      601 tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttcgaat
      661 tcggcagcac ttttctttca ttggggggaa ggcgtgagga aagtaccaaa cagcagcaga
      721 cttttaaact ttaaacagac aggtctgaga gcctgaactc tcatttttcc tttgacttca
      781 gcctccaagg agttccacca actttggcgc gccggcttca ctttcattaa gtgaaagaga
      841 ggtgcccaga catgggtgac tggagtgcct tggggaagct tctggacaag gtccaagcct
      901 actccaccgc tggagggaag gtgtggctgt cagtgctctt catattcaga atcctgctcc
      961 tggggacagc tgttgagtca gcttggggtg atgaacagtc tgcctttcgc tgtaacactc
     1021 aacaacctgg ctgcgaaaac gtctgctatg acaagtcctt ccccatctct cacgtgcgct
     1081 tctgggtcct tcagatcata ttcgtgtctg tgcccacact cctgtacttg gcccatgtgt
     1141 tctatgtgat gaggaaggaa gagaagctaa acaagaaaga agaggagctc aaagtggccc
     1201 agactgacgg ggtcaacgtg gagatgcacc tgaagcagat tgaaatcaag aagttcaagt
     1261 acgggattga agagcacggc aaggtgaaaa tgaggggcgg cttgctgaga acctacatca
     1321 tcagcatcct cttcaagtct gtcttcgagg tggccttcct gctcatccag tggtacatct
     1381 atgggttcag cttgagcgcg gtctacacct gcaagagaga tccctgcccc caccaggtag
     1441 actgcttcct ctcacgtccc acggagaaaa ccatcttcat catcttcatg ctggtggtgt
     1501 ccttggtgtc tctcgctttg aacatcattg agctcttcta cgtcttcttc aagggcgtta
     1561 aggatcgcgt gaagggaaga agcgatcctt accacgccac cactggccca ctgagcccat
     1621 caaaagactg cggatctcca aaatacgcct acttcaatgg ctgctcctca ccaacggctc
     1681 cactctcgcc tatgtctcct cctgggtaca agctggttac tggtgacaga aacaattcct
     1741 cgtgccgcaa ttacaacaag caagctagcg agcaaaactg ggcgaactac agcgcagagc
     1801 aaaatcgcat ggggcaggcc ggaagcacca tctccaactc gcacgcccag ccgttcgatt
     1861 tccccgacga caaccagaat gccaaaaaag ttgctgctgg acatgaactc cagccattag
     1921 ccatcgtgga ccaacgacct tccagcagag ccagcagccg cgccagcagc aggcctcggc
     1981 ctgatgacct ggagattgcg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg
     2041 ataacatggc catcattaag gagttcatgc gcttcaaggt gcacatggag gggtccgtga
     2101 acggccacgt gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     2161 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cacctgggac atcctgagcc
     2221 ctcagttcat gtacggctcc aatgcctacg tgaagcaccc cgccgacatc cccgactact
     2281 ttaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaaattc gaggacggcg
     2341 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     2401 agctgcgcgg caccaacttc ccctccgacg gccccgtgat gcagaagaag accatgggct
     2461 gggaggccct ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gaggtcaagc
     2521 ccagagtgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     2581 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa ccgcaagctg gacatcacca
     2641 gccacaacga ggactacacc atcgtggagc agtacgagag agccgagggc cgccactcca
     2701 ccggcggcat ggacgagctg tacaagggcg gcatggagaa gagcttcgtg atcaccgacc
     2761 cccgcctgcc cgacaacccc atcatcttcg ccagcgacgg cttcctggag ctgaccgagt
     2821 acagcagaga ggagatcctg ggccgcaacg gccgcttcct gcagggcccc gagaccgacc
     2881 aggccaccgt gcagaagatc cgcgacgcca tccgcgacca gcgcgagatc accgtgcagc
     2941 tgatcaacta caccaagagc ggcaagaagt tctggaacct gctgcacctg cagcccatgc
     3001 gcgaccagaa gggcgagctg cagtacttca tcggcgtgca gctggacggc taagcggccg
     3061 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3121 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3181 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3241 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3301 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3361 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3421 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3481 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3541 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3601 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3661 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3721 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3781 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3841 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3901 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3961 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4021 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4081 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4141 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4201 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4261 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4321 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4381 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4441 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4501 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4561 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4621 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4681 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4741 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4801 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4861 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4921 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4981 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5041 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5101 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5161 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5221 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5281 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5341 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5401 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5461 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5521 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5581 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5701 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5761 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5821 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5881 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5941 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6001 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6061 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6121 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6181 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6241 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6301 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattc
//
