LOCUS       Exported                5504 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  In vivo visualization of actin cytoskeleton (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    mNeonGreen-UtrCH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv 160374
REFERENCE   2  (bases 1 to 5504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5504
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     224..244
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(269..373)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      400..855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(487..506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     697..718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    862..983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(873..892)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     927..946
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    1107..1159
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(1955..2662)
                     /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      2398..2407
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      2659..2668
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(2707..2910)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2736..2756)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2911..3214
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3389..3977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3469..3488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4306..4353
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4378..4397
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4585..5379)
                     /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(4696..4715)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5306..5325
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5414..267)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5428..59
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5482..5501)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
       61 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
      121 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
      181 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
      241 ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt
      301 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
      361 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
      421 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      481 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      541 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      601 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      661 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      721 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      781 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      841 taacgcttac aatttacgcc ttaagataca ttgatgagtt tggacaaacc acaactagaa
      901 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
      961 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1021 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     1081 attatgatca gttatctaga tccggtggat cccgggcccg cggtaccgtc gactgcagaa
     1141 ttcgaagctt gagctcgagt tagtctatgg tgacttgctg aggtagcacc tcaaacaaag
     1201 atgttaaata cataattatg gatttcttgt caggaagccg aacggcaaca tcttcaggat
     1261 ctaacagctt ttcaattccc aaataagttt gagccttgct gaaggcatgt tcaagtctct
     1321 caattggtga cattttgaca actttatccc agctgaagag atcaggttta tgtcggtgga
     1381 ggacagcatt aaaggcgagt ccatctgtcc agctggtggt gaagttgagg acgttgactt
     1441 ggctgtaggg cctggtggtc tgacgcaccc agctgagcag gatcttctca ctgttcgtct
     1501 gctgcaggtc cgacatgaca tccttcatga catctttcac ctgccagtgc aaaatgatgc
     1561 tccaaagtaa ccccaaagtc agtttgtgat ttccatccac aatgtcggtt ccccctatat
     1621 tcactaattc cacattgttc tgatgtaaaa cctgcagcac tctgttgacg ttatttaagg
     1681 catgtaccct tgtggaacca cgttcctttg gcagtgatgt tcctgtgagg ccttctagaa
     1741 gatccaatag cttccttcca tctttgaggt ctgtgaacat atcattgatg ggtggtttcc
     1801 cactctttga aaatcgagca tttatccatt tggtaaaggt tttcttctgt acgtcattgt
     1861 gttcatcaga tctggactta atgatatcac tgaattcgtt ctgcccattg tcaggactgg
     1921 cttcatgttc tccatacttg gccatggttc cggacttgta cagctcgtcc atgcccatca
     1981 catcggtaaa ggccttttgc cactccttga agttgagctc ggtcttggag tgcttgagct
     2041 ccgtcttacg gaacacgtac atcggctggt tcttcagata gttagccgcc attggcttgg
     2101 caaaggtgta ggtggtccgc gcagtgctcc ggtagcgctt gccatttcca gtggtgtaac
     2161 tccacttaaa ggtactgatg atggttttgt cgttggggta agtcttcttc gacctgcacc
     2221 agtccgcagc ggtcagcgag ttggtcatca caggaccgtc agcagggaaa ccagtcccct
     2281 tcacctgggc ctctcctttg atgtggcttc cctcgtaggt gtagcggtag ttaacagtaa
     2341 gggaggcacc atcttcaaac tgcattgtgc gatggacttg gtagccggag ccatctacca
     2401 tggcggcctg gaaaggcgac atcccgtcag ggtagggcag gtactgatgg aagccatacc
     2461 cgatatgagg gaccagaatc cagggggaga actggaggtc acccttggtg gacttcaggt
     2521 ttaactcctc ataaccatca tttggattgc cggtgccctg acccaccatg tcaaagtcca
     2581 caccgttgat ggagccaaag atgtgtaact catgtgtcgc tgggagagag gccatgttat
     2641 cctcctcgcc cttgctcacc atggtggcga ccggtagcgc tagcggatct gacggttcac
     2701 taaaccagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     2761 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     2821 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     2881 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     2941 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     3001 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     3061 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     3121 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     3181 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     3241 taatgacccc gtaattgatt actattaata actaatgcat ggcggtaata cggttatcca
     3301 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taacctgagg
     4141 ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc tgggccttca cccgaacttg
     4201 gggggtgggg tggggaaaag gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg
     4261 gggtatcgac agagtgccag ccctgggacc gaaccccgcg tttatgaaca aacgacccaa
     4321 caccgtgcgt tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg
     4381 tctccttccg tgtttcagtt agcctccccc tagggtgggc gaagaactcc agcatgagat
     4441 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     4501 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
     4561 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     4621 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     4681 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     4741 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     4801 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag
     4861 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     4921 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     4981 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     5041 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     5101 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     5161 cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga
     5221 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     5281 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     5341 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     5401 ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     5461 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaa
//
