LOCUS       sequence-191169-        4805 bp    DNA     circular SYN 14-APR-2025
DEFINITION  In vivo visualization of actin cytoskeleton (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4805)
  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 4805)
  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: "bioRxiv 
            160374"
COMMENT     Sequence Label: ITPKA-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..4805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..257
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     214..234
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(259..363)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      390..845
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(477..496)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     687..708
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    852..973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(863..882)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     917..936
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1096..1806)
                     /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      1542..1551
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1803..1812
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1998..2201)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2027..2047)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2202..2505
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2680..3268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2760..2779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3597..3644
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3669..3688
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3876..4670)
                     /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             complement(3876..4670)
                     /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(3987..4006)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4597..4616
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4705..4805,1..257))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4719..4805,1..49)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
       61 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
      121 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
      181 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
      241 ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta
      301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      361 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
      421 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
      481 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
      541 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
      601 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
      661 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
      721 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
      781 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
      841 aatttacgcc ttaagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
      901 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
      961 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1021 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatct
     1081 agagtcgcgg ccgctttact tgtacagctc gtccatgccc atcacatcgg taaaggcctt
     1141 ttgccactcc ttgaagttga gctcggtctt ggagtgcttg agctccgtct tacggaacac
     1201 gtacatcggc tggttcttca gatagttagc cgccattggc ttggcaaagg tgtaggtggt
     1261 ccgcgcagtg ctccggtagc gcttgccatt tccagtggtg taactccact taaaggtact
     1321 gatgatggtt ttgtcgttgg ggtaagtctt cttcgacctg caccagtccg cagcggtcag
     1381 cgagttggtc atcacaggac cgtcagcagg gaaaccagtc cccttcacct gggcctctcc
     1441 tttgatgtgg cttccctcgt aggtgtagcg gtagttaaca gtaagggagg caccatcttc
     1501 aaactgcatt gtgcgatgga cttggtagcc ggagccatct accatggcgg cctggaaagg
     1561 cgacatcccg tcagggtagg gcaggtactg atggaagcca tacccgatat gagggaccag
     1621 aatccagggg gagaactgga ggtcaccctt ggtggacttc aggtttaact cctcataacc
     1681 atcatttgga ttgccggtgc cctgacccac catgtcaaag tccacaccgt tgatggagcc
     1741 aaagatgtgt aactcatgtg tcgctgggag agaggccatg ttatcctcct cgcccttgct
     1801 caccatggtg gcgaccggtg gatccgatcc agatccgccc gcttcgaaga gcaggcgcag
     1861 ctccccgacg ctccggcgcg gagcccgctc caacccgggg ctgcagggcc ccgcgccccg
     1921 tggtcgcgcc atgcccataa gcttgagctc gagatctgag tccggtagcg ctagcggatc
     1981 tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc
     2041 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
     2101 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     2161 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     2221 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     2281 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     2341 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     2401 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     2461 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     2521 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactaatgca tggcggtaat
     2581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3421 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
     3481 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
     3541 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     3601 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     3661 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     3721 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     3781 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     3841 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     3901 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     3961 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     4021 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     4081 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     4141 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     4201 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     4261 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     4321 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     4381 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     4441 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     4501 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     4561 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     4621 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     4681 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     4741 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     4801 tagtc
//
