LOCUS       sequence-252343-        4883 bp    DNA     circular SYN 14-APR-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 Apr 14, 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        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      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1611..1732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1648..1667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1702..1721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1739..2194)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1876..1897)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2088..2107
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2221..2325
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2327..2684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2327..2518
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2350..2370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2535..2670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2719..3513
                     /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             2719..3513
                     /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(2773..2792)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3383..3402
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3701..3720)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3745..3792
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4121..4709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4610..4629
                     /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 gctagcgcca
      541 ccatggtcag caaaggcgaa gaagacaaca tggcatcact gccagcgaca cacgaacttc
      601 atattttcgg ctccatcaat ggtgtggact tcgatatggt tggacagggt actgggaacc
      661 ctaacgatgg atatgaggag ttgaacttga agagtaccaa gggcgatctt cagttttcac
      721 cttggattct cgtacctcat ataggttatg gctttcatca atatctgccc tatcctgacg
      781 gtatgtcacc tttccaagca gctatggttg acgggagcgg ttatcaggtt caccggacaa
      841 tgcagtttga agatggagcc tctttgaccg taaattatcg gtatacttat gagggcagcc
      901 atattaaagg cgaggctcaa gttaagggga ctgggttccc cgcggatggt cccgtaatga
      961 caaacagtct gacggctgca gactggtgcc gctctaagaa gacatatcca aatgacaaga
     1021 cgattattag cactttcaag tggtcataca ctacaggaaa tggtaaacgc tataggagca
     1081 ctgcccggac cacgtatact ttcgcgaagc cgatggcggc aaattatttg aaaaaccaac
     1141 cgatgtatgt gttcagaaag acagaactca agcattctaa aactgaactg aacttcaaag
     1201 agtggcagaa agcctttact gatgtaatgg gcatggacga actctataag tccggactca
     1261 gatccggtgg aggcggttca ggcggaggtg gctctggcgg tggcggatcg agatctaaac
     1321 taattgctaa aagtgcagaa gacgaaaaag cgaaggaaga accagggaac catacgacgt
     1381 taattcttgc aatgttagct attggcgtgt tctctttagg ggcgtttatc aaaattattc
     1441 aattaagaaa aaataattaa gtcgacggta ccgcgggccc gggatccacc ggatctagat
     1501 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     1561 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     1621 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1681 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     1741 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     1801 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     1861 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     1921 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     1981 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2041 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2101 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2161 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2221 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2281 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     2341 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2401 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2521 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2581 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2641 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     2701 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2761 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2821 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2881 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     2941 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3001 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3061 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3121 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3181 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3241 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3301 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3361 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3421 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3481 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3541 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3601 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3661 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     3721 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     3781 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3841 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3901 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     3961 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4021 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4081 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4141 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4201 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4261 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4321 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4381 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4441 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4501 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4561 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4621 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4681 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4741 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4801 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     4861 catagcccat atatggagtt ccg
//
