LOCUS       Exported                6251 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  In vivo visualization of F-actin (can be used for colocalization 
            studies).
ACCESSION   .
VERSION     .
KEYWORDS    ITPKA_3xmNeonGreen
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6251)
  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 6251)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6251
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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      754..763
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             760..1464
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      1015..1024
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1483..2187
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELY"
     regulatory      1738..1747
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2206..2916
                     /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      2461..2470
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    3039..3160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3076..3095)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3130..3149
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3167..3622)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3304..3325)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3516..3535
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3649..3753
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3755..4112
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3778..3798)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3963..4098
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4025..4044
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4147..4941
                     /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(4201..4220)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4811..4830
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5129..5148)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5173..5220
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5549..6137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6038..6057
                     /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 ccggactcag atctcgagct caagcttatg ggcatggcgc gaccacgggg cgcggggccc
      661 tgcagccccg ggttggagcg ggctccgcgc cggagcgtcg gggagctgcg cctgctcttc
      721 gaagcgggcg gatctggatc ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
      781 gataacatgg cctctctccc agcgacacat gagttacaca tctttggctc catcaacggt
      841 gtggactttg acatggtggg tcagggcacc ggcaatccaa atgatggtta tgaggagtta
      901 aacctgaagt ccaccaaggg tgacctccag ttctccccct ggattctggt ccctcatatc
      961 gggtatggct tccatcagta cctgccctac cctgacggga tgtcgccttt ccaggccgcc
     1021 atggtagatg gctccggcta ccaagtccat cgcacaatgc agtttgaaga tggtgcctcc
     1081 cttactgtta actaccgcta cacctacgag ggaagccaca tcaaaggaga ggcccaggtg
     1141 aaggggactg gtttccctgc tgacggtcct gtgatgacca actcgctgac cgctgcggac
     1201 tggtgcaggt cgaagaagac ttaccccaac gacaaaacca tcatcagtac ctttaagtgg
     1261 agttacacca ctggaaatgg caagcgctac cggagcactg cgcggaccac ctacaccttt
     1321 gccaagccaa tggcggctaa ctatctgaag aaccagccga tgtacgtgtt ccgtaagacg
     1381 gagctcaagc actccaagac cgagctcaac ttcaaggagt ggcaaaaggc ctttaccgat
     1441 gtgatgggca tggacgagct gtaccagggt ggcagcggtg gcatggtgag caagggcgag
     1501 gaggataaca tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac
     1561 ggtgtggact ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag
     1621 ttaaacctga agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat
     1681 atcgggtatg gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc
     1741 gccatggtag atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc
     1801 tcccttactg ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag
     1861 gtgaagggga ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg
     1921 gactggtgca ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag
     1981 tggagttaca ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc
     2041 tttgccaagc caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag
     2101 acggagctca agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc
     2161 gatgtgatgg gcatggacga gctgtaccag ggtggcagcg gtggcatggt gagcaagggc
     2221 gaggaggata acatggcctc tctcccagcg acacatgagt tacacatctt tggctccatc
     2281 aacggtgtgg actttgacat ggtgggtcag ggcaccggca atccaaatga tggttatgag
     2341 gagttaaacc tgaagtccac caagggtgac ctccagttct ccccctggat tctggtccct
     2401 catatcgggt atggcttcca tcagtacctg ccctaccctg acgggatgtc gcctttccag
     2461 gccgccatgg tagatggctc cggctaccaa gtccatcgca caatgcagtt tgaagatggt
     2521 gcctccctta ctgttaacta ccgctacacc tacgagggaa gccacatcaa aggagaggcc
     2581 caggtgaagg ggactggttt ccctgctgac ggtcctgtga tgaccaactc gctgaccgct
     2641 gcggactggt gcaggtcgaa gaagacttac cccaacgaca aaaccatcat cagtaccttt
     2701 aagtggagtt acaccactgg aaatggcaag cgctaccgga gcactgcgcg gaccacctac
     2761 acctttgcca agccaatggc ggctaactat ctgaagaacc agccgatgta cgtgttccgt
     2821 aagacggagc tcaagcactc caagaccgag ctcaacttca aggagtggca aaaggccttt
     2881 accgatgtga tgggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     2941 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3001 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3061 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3121 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     3181 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3241 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3301 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3361 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3421 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3481 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3541 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3601 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3661 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3721 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3781 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3841 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3901 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3961 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4021 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4081 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4141 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4201 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4261 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4321 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4381 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4441 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4501 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4561 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4621 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4681 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4741 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4801 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4861 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4921 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4981 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5041 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5101 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5161 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5221 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5281 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5341 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5401 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6001 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6061 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6181 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6241 ccgccatgca t
//
