LOCUS       sequence-245169-        4883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a mammalian expression vector for mitochondria-targeting humanized 
            mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4883)
  AUTHORS   Tanida-Miyake E, Koike M, Uchiyama Y, Tanida I
  TITLE     Optimization of mNeonGreen for Homo sapiens increases its 
            fluorescent intensity in mammalian cells.
  JOURNAL   PLoS One. 2018 Jan 17;13(1):e0191108. doi: 
            10.1371/journal.pone.0191108. eCollection 2018.
  PUBMED    29342181
REFERENCE   2  (bases 1 to 4883)
  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 One.";
            date: "2018-01-17"; volume: "13(1):e0191108. doi"; pages: " 
            10.1371/journal.pone.0191108. eCollection 2018"
COMMENT     Sequence Label: pmNeonGreenHO-mito
FEATURES             Location/Qualifiers
     source          1..4883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1671..1792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1708..1727)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1762..1781
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1799..2254)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1936..1957)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2148..2167
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2281..2385
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2387..2744
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2387..2578
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2410..2430)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2595..2730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2779..3573
                     /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             2779..3573
                     /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(2833..2852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3443..3462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3761..3780)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3805..3852
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4181..4769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4670..4689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatggtcag caaaggcgaa gaagacaaca tggcatcact gccagcgaca cacgaacttc
      661 atattttcgg ctccatcaat ggtgtggact tcgatatggt tggacagggt actgggaacc
      721 ctaacgatgg atatgaggag ttgaacttga agagtaccaa gggcgatctt cagttttcac
      781 cttggattct cgtacctcat ataggttatg gctttcatca atatctgccc tatcctgacg
      841 gtatgtcacc tttccaagca gctatggttg acgggagcgg ttatcaggtt caccggacaa
      901 tgcagtttga agatggagcc tctttgaccg taaattatcg gtatacttat gagggcagcc
      961 atattaaagg cgaggctcaa gttaagggga ctgggttccc cgcggatggt cccgtaatga
     1021 caaacagtct gacggctgca gactggtgcc gctctaagaa gacatatcca aatgacaaga
     1081 cgattattag cactttcaag tggtcataca ctacaggaaa tggtaaacgc tataggagca
     1141 ctgcccggac cacgtatact ttcgcgaagc cgatggcggc aaattatttg aaaaaccaac
     1201 cgatgtatgt gttcagaaag acagaactca agcattctaa aactgaactg aacttcaaag
     1261 agtggcagaa agcctttact gatgtaatgg gcatggacga actctataag tccggactca
     1321 gatccggtgg aggcggttca ggcggaggtg gctctggcgg tggcggatcg agatctaaac
     1381 taattgctaa aagtgcagaa gacgaaaaag cgaaggaaga accagggaac catacgacgt
     1441 taattcttgc aatgttagct attggcgtgt tctctttagg ggcgtttatc aaaattattc
     1501 aattaagaaa aaataattaa gtcgacggta ccgcgggccc gggatccacc ggatctagat
     1561 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     1621 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     1681 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1741 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     1801 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     1861 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     1921 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     1981 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2041 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2101 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2161 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2221 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2281 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2341 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     2401 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2461 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2521 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2581 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2641 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2701 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     2761 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2821 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2881 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2941 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     3001 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3061 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3121 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3181 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3241 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3301 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3361 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3421 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3481 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3541 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3601 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3661 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3721 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     3781 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     3841 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3901 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3961 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     4021 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4081 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4141 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4201 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4261 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4321 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4381 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4441 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4501 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4561 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4621 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4681 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4741 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4801 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4861 ataaccgtat taccgccatg cat
//
