LOCUS       sequence-254785-        4333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 448 / 473, Emission: 
            500 / 528.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4333)
  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 4333)
  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-N1
FEATURES             Location/Qualifiers
     source          1..4333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    589..669
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             677..997
                     /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    1119..1240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1156..1175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1210..1229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1247..1702)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1384..1405)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1596..1615
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1729..1833
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1835..2192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1835..2026
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1858..1878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2043..2178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2227..3021
                     /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             2227..3021
                     /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(2281..2300)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2891..2910
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3209..3228)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3253..3300
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3629..4217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4118..4137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      601 ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga
      661 tccaccggtc gccaccatgg agaaaagttt cgtgataact gatccacggc tgccagacaa
      721 tcccatcatc ttcgcatccg atggcttcct ggagctgacc gagtattcca gagaggagat
      781 cctgggccgc aatggccgct ttctgcaggg accagagaca gaccaggcca cagtgcagaa
      841 gattcgcgat gccattagag atcagcgcga gattaccgtg cagctgataa actacacaaa
      901 aagcgggaag aaattctgga acctcctgca cctccagccc atgagggacc agaagggtga
      961 gctccagtat ttcatcggag tgcagctgga tggataagcg gccgcgactc tagatcataa
     1021 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1081 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1141 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1201 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     1261 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1321 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1381 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1441 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1501 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1561 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     1621 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     1681 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1741 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1801 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     1861 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     1921 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     1981 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2041 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2101 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2161 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     2221 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2281 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2341 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2401 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2461 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2521 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2581 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2641 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     2701 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     2761 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     2821 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     2881 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     2941 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3001 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3061 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3121 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3181 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3241 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     3301 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     3361 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     3421 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     3481 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3541 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3601 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3661 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3721 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3781 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3841 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3901 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3961 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4021 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4081 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4141 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4201 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4261 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4321 ccgccatgca tta
//
