LOCUS       sequence-193754-        7024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Alpha Actinin, Excitation: 448 / 473, Emission: 500 / 
            528.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7024)
  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 7024)
  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: miniSOG-Alpha-Actinin-19
FEATURES             Location/Qualifiers
     source          1..7024
                     /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      3304..3313
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3310..3630
                     /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    3752..3873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3789..3808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3843..3862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3880..4335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4017..4038)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4229..4248
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4362..4466
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4468..4825
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4468..4659
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4491..4511)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4676..4811
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4860..5654
                     /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             4860..5654
                     /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(4914..4933)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5524..5543
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5842..5861)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5886..5933
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6262..6850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6751..6770
                     /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 caagcttcgc accatcatgg accattatga ttctcagcaa
      601 accaacgatt acatgcagcc agaagaggac tgggaccggg acctgctcct ggacccggcc
      661 tgggagaagc agcagagaaa gacattcacg gcatggtgta actcccacct ccggaaggcg
      721 gggacacaga tcgagaacat cgaagaggac ttccgggatg gcctgaagct catgctgctg
      781 ctggaggtca tctcaggtga acgcttggcc aagccagagc gaggcaagat gagagtgcac
      841 aagatctcca acgtcaacaa ggccctggat ttcatagcca gcaaaggcgt caaactggtg
      901 tccatcggag ccgaagaaat cgtggatggg aatgtgaaga tgaccctggg catgatctgg
      961 accatcatcc tgcgctttgc catccaggac atctccgtgg aagagacttc agccaaggaa
     1021 gggctgctcc tgtggtgtca gagaaagaca gccccttaca aaaatgtcaa catccagaac
     1081 ttccacataa gctggaagga tggcctcggc ttctgtgctt tgatccaccg acaccggccc
     1141 gagctgattg actacgggaa gctgcggaag gatgatccac tcacaaatct gaatacggct
     1201 tttgacgtgg cagagaagta cctggacatc cccaagatgc tggatgccga agacatcgtt
     1261 ggaactgccc gaccggatga gaaagccatc atgacttacg tgtctagctt ctaccacgcc
     1321 ttctctggag cccagaaggc ggagacagca gccaatcgca tctgcaaggt gttggccgtc
     1381 aaccaggaga acgagcagct tatggaagac tacgagaagc tggccagtga tctgttggag
     1441 tggatccgcc gcacaatccc gtggctggag aaccgggtgc ccgagaacac catgcatgcc
     1501 atgcaacaga agctggagga cttccgggac taccggcgcc tgcacaagcc gcccaaggtg
     1561 caggagaagt gccagctgga gatcaacttc aacacgctgc agaccaagct gcggctcagc
     1621 aaccggcctg ccttcatgcc ctctgagggc aggatggtct cggacatcaa caatgcctgg
     1681 ggctgcctgg agcaggtgga gaagggctat gaggagtggt tgctgaatga gatccggagg
     1741 ctggagcgac tggaccacct ggcagagaag ttccggcaga aggcctccat ccacgaggcc
     1801 tggactgacg gcaaagaggc catgctgcga cagaaggact atgagaccgc caccctctcg
     1861 gagatcaagg ccttgctcaa gaagcatgag gccttcgaga gtgacctggc tgcccaccag
     1921 gaccgtgtgg agcagattgc cgccatcgca caggagctca atgagctgga ctattatgac
     1981 tcacccagtg tcaacgcccg ttgccaaaag atctgtgacc agtgggacaa tctgggggcc
     2041 ctaactcaga agcgaaggga agctctggag cggaccgaga aactgctgga gaccattgac
     2101 cagctgtact tggagtatgc caagcgggct gcacccttca acaactggat ggagggggcc
     2161 atggaggacc tgcaggacac cttcattgtg cacaccattg aggagatcca gggactgacc
     2221 acagcccatg agcagttcaa ggccaccctc cctgatgccg acaaggagcg cctggccatc
     2281 ctgggcatcc acaatgaggt gtccaagatt gtccagacct accacgtcaa tatggcgggc
     2341 accaacccct acacaaccat cacgcctcag gagatcaatg gcaaatggga ccacgtgcgg
     2401 cagctggtgc ctcggaggga ccaagctctg acggaggagc atgcccgaca gcagcacaat
     2461 gagaggctac gcaagcagtt tggagcccag gccaatgtca tcgggccctg gatccagacc
     2521 aagatggagg agatcgggag gatctccatt gagatgcatg ggaccctgga ggaccagctc
     2581 agccacctgc ggcagtatga gaagagcatc gtcaactaca agccaaagat tgatcagctg
     2641 gagggcgacc accagctcat ccaggaggcg ctcatcttcg acaacaagca caccaactac
     2701 accatggagc acatccgtgt gggctgggag cagctgctca ccaccatcgc caggaccatc
     2761 aatgaggtag agaaccagat cctgacccgg gatgccaagg gcatcagcca ggagcagatg
     2821 aatgagttcc gggcctcctt caaccacttt gaccgggatc actccggcac actgggtccc
     2881 gaggagttca aagcctgcct catcagcttg ggttatgata ttggcaacga cccccaggga
     2941 gaagcagaat ttgcccgcat catgagcatt gtggacccca accgcctggg ggtagtgaca
     3001 ttccaggcct tcattgactt catgtcccgc gagacagccg acacagatac agcagaccaa
     3061 gtcatggctt ccttcaagat cctggctggg gacaagaact acattaccat ggacgagctg
     3121 cgccgcgagc tgccacccga ccaggctgag tactgcatcg cgcggatggc cccctacacc
     3181 ggccccgact ccgtgccagg tgctctggac tacatgtcct tctccacggc gctgtacggc
     3241 gagagtgacc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     3301 gtcgccacca tggagaaaag tttcgtgata actgatccac ggctgccaga caatcccatc
     3361 atcttcgcat ccgatggctt cctggagctg accgagtatt ccagagagga gatcctgggc
     3421 cgcaatggcc gctttctgca gggaccagag acagaccagg ccacagtgca gaagattcgc
     3481 gatgccatta gagatcagcg cgagattacc gtgcagctga taaactacac aaaaagcggg
     3541 aagaaattct ggaacctcct gcacctccag cccatgaggg accagaaggg tgagctccag
     3601 tatttcatcg gagtgcagct ggatggataa gcggccgcga ctctagatca taatcagcca
     3661 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3721 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3781 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3841 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     3901 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3961 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4021 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4081 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4141 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4201 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4261 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4321 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4381 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4441 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     4501 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4561 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4621 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4681 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4741 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4801 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     4861 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4921 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4981 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5041 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5101 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5161 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5221 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5281 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5341 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     5401 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5461 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5521 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5581 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5641 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5701 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5761 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5821 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5881 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     5941 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6001 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     6061 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     6121 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6181 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6241 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6301 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6361 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6421 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6481 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6541 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6601 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6661 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6721 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6781 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6841 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6901 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6961 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     7021 tccg
//
