LOCUS       sequence-478807-        7500 bp    DNA     circular SYN 16-JUL-2025
DEFINITION  Expresses mNeonGreen -NUP98 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7500)
  AUTHORS   Zhang H, Lesnov GD, Subach OM, Zhang W, Kuzmicheva TP, Vlaskina AV, 
            Samygina VR, Chen L, Ye X, Nikolaeva AY, Gabdulkhakov A, Papadaki S,
            Qin W, Borshchevskiy V, Perfilov MM, Gavrikov AS, Drobizhev M, 
            Mishin AS, Piatkevich KD, Subach FV
  TITLE     Bright and stable monomeric green fluorescent protein derived from 
            StayGold.
  JOURNAL   Nat Methods. 2024 Apr;21(4):657-665. doi: 
            10.1038/s41592-024-02203-y. Epub 2024 Feb 26.
  PUBMED    38409224
REFERENCE   2  (bases 1 to 7500)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-024-02203-y"; journalName: "Nat Methods"; date: 
            "2024-04"; volume: "21"; issue: "4"; pages: "657-665"
COMMENT     Sequence Label: pmNeonGreen-NUP98
FEATURES             Location/Qualifiers
     source          1..7500
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      868..877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4288..4409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4325..4344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4379..4398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4416..4871)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4553..4574)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4765..4784
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4898..5002
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5004..5361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5004..5195
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5027..5047)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5212..5347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5396..6190
                     /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             5396..6190
                     /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(5450..5469)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6060..6079
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6378..6397)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6422..6469
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6798..7386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7287..7306
                     /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 gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggcctctct cccagcgaca
      661 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
      721 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
      781 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
      841 taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg ctaccaagtc
      901 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
      961 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     1021 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     1081 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccacgggaaa tggcaagcgc
     1141 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     1201 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     1261 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     1321 tccggccgga ctcagatctc gagcatgttt aacaaatcat ttggaacacc ctttgggggt
     1381 ggcacaggtg gctttggcac aacttcaaca tttggacaga atactggctt tggcactact
     1441 agtggagggg catttggaac atctgcattt ggttctagca acaatactgg aggcctcttt
     1501 ggaaattcac agactaaacc aggaggattg tttggaacca gttcatttag ccagccagct
     1561 acctccacaa gcactggctt tgggtttggt acgtcaacag gaacagcaaa taccttgttt
     1621 ggaactgcaa gcacagggac cagtctcttc tcatcccaaa acaatgcctt tgcacaaaat
     1681 aaaccaactg gctttggcaa ttttggaacc agtactagca gtggaggact ctttggaacc
     1741 acaaatacca cctctaatcc ttttggcagc acatctggct ccctctttgg gccaagtagt
     1801 tttacagctg ctcctactgg gactactatt aaatttaacc ctccaactgg tacagatact
     1861 atggtcaaag ctggagttag cactaacata agtaccaagc accagtgtat tactgctatg
     1921 aaagaatatg aaagcaagtc actagaggaa cttcgtttag aggattatca ggctaacagg
     1981 aagggcccac agaaccaggt gggagcaggt accacaactg gcttgtttgg gtcttctcca
     2041 gccacttcca gcgcaacagg actcttcagc tcctccacca ctaattcagg ctttgcatat
     2101 ggtcagaaca aaactgcctt tggaactagt acaactggat ttggaacaaa tccaggtggt
     2161 ctctttggcc aacagaatca gcagactacc agcctcttca gcaaaccatt tggccaggct
     2221 acaaccaccc agaacactgg cttttccttt ggtaatacca gcaccatagg acagccaagc
     2281 accaacacca tgggattatt tggagtaacc caagcctcac agcctggagg tctttttggg
     2341 acagctacaa acaccagcac tgggacagca tttggaacag gaacaggtct ctttgggcag
     2401 accaatactg gatttggtgc tgttggttcg accctgtttg gcaataacaa gcttactaca
     2461 tttggaagca gcacaaccag tgcaccttca tttggtacaa ccagtggcgg gctctttggt
     2521 aacaaaccaa ccctgacttt aggaaccaat acaaacactt ctaattttgg ttttggcaca
     2581 aataccagtg ggaatagtat ttttggaagt aaaccagcac ctgggactct tggaactggg
     2641 cttggtgcag gatttggaac agctcttggt gctggacagg catctttgtt tgggaacaac
     2701 caacctaaga ttggagggcc tcttggtaca ggagcctttg gggcccctgg atttaatact
     2761 acgacagcca ctttgggctt tggagccccc caggccccag tagctttgac agatccaaat
     2821 gcttctgctg cccagcaggc tgttctccag cagcacatca atagtctaac atactcacct
     2881 tttggagact ctcctctctt ccggaatccg atgtcagacc ctaagaagaa ggaagagaga
     2941 ttgaaaccaa caaatccagc agcccagaag gctcttacta cacctactca ttataaactg
     3001 acaccccgcc ctgccactag agtccggcca aaggctttac aaacaacagg cacagccaag
     3061 tcacatctct ttgatgggct ggatgacgat gaaccatccc tagccaatgg agcattcatg
     3121 cccaagaaga gcattaagaa gttggttttg aagaacctta ataatagcaa tctcttttct
     3181 cctgttaatc gtgattcaga aaatctagct tcaccatctg aatatccaga aaatggagag
     3241 agatttagtt tcctaagcaa acctgttgat gagaatcacc agcaggatgg agatgaagat
     3301 tcccttgttt cacattttta tactaaccct attgccaaac ctattcctca aaccccagaa
     3361 agtgctggaa ataaacacag caacagcaac agtgtggatg ataccattgt tgcattaaac
     3421 atgcgtgctg ctttgcgaaa tgggctggaa ggaagcagtg aagaaacgtc ttttcatgat
     3481 gagtcacttc aggatgaccg agaagaaata gaaaataatt cttaccatat gcacccagca
     3541 ggtattattc tcactaaggt tggttactat actattccat ctatggatga ccttgctaaa
     3601 attaccaatg aaaaaggaga gtgcattgtc tctgatttca ctattggtcg gaaaggttat
     3661 ggttcaatct attttgaagg agatgtgaat ttgacaaatc taaatttgga tgatattgtg
     3721 catatccgga ggaaagaagt agttgtctac ttagatgata accaaaaacc acctgtgggt
     3781 gaagggctaa ataggaaggc tgaagttaca ttggatggag tttggccaac agataaaaca
     3841 tctcgttgtt taataaagag cccagatcgc cttgctgata tcaactatga aggaagattg
     3901 gaagcagttt caaggaaaca gggagctcaa ttcaaagaat accggcctga aactggttct
     3961 tgggtgttta aggtctccca tttttctaag tatggccttc aggattctga tgaagaggag
     4021 gaggagcatc cgtctaaaac tagtacaaag aagttgaaga ctgctccttt gcctcctgca
     4081 agccagacta cgcccttgca gatggctctt aatggcaaac ctgcacctcc acctcaggta
     4141 gagaaaaaag gacagtgagg atccaccgga tctagataac tgatcataat cagccatacc
     4201 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4261 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4321 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4381 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     4441 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4501 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4561 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4621 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4681 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4741 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4801 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4861 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4921 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4981 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     5041 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5101 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5161 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5221 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5281 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5341 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     5401 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5461 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5521 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     5581 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5641 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5701 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5761 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5821 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5881 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5941 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     6001 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     6061 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6121 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6181 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6241 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6301 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6361 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     6421 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     6481 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     6541 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     6601 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     6661 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6721 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6781 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6841 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6901 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6961 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7021 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7081 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7141 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7201 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7261 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7321 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7381 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7441 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
