LOCUS       sequence-248811-        6410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of focal adhesions (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6410)
  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 (2020) 160374; doi: https://doi.org/10.1101/160374
REFERENCE   2  (bases 1 to 6410)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            (2020) 160374; doi: https://doi.org/10.1101/160374"
COMMENT     Sequence Label: mNeongreen-PXN
FEATURES             Location/Qualifiers
     source          1..6410
                     /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    3198..3319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3235..3254)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3289..3308
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3326..3781)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3463..3484)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3675..3694
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3808..3912
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3914..4271
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3914..4105
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3937..3957)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4122..4257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4306..5100
                     /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             4306..5100
                     /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(4360..4379)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4970..4989
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5288..5307)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5332..5379
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5708..6296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6197..6216
                     /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 ccactggaaa tggcaagcgc
     1141 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     1201 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     1261 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagc tgcggccgcc ggatccaagc ttatggacga cctcgacgcc
     1381 ctgctggcgg acttggagtc taccacctcc cacatctcca aacggcctgt gttcttgtcg
     1441 gaggagaccc cctactcata cccaactgga aaccacacat accaggagat tgccgtgcca
     1501 ccccccgtcc ccccaccccc gtccagcgag gccctcaatg gcacaatcct tgacccctta
     1561 gaccagtggc agcccagcgg ctcccgattc atccaccagc agcctcagtc ctcatcacct
     1621 gtgtacggct ccagtgccaa aacttccagt gtctccaacc ctcaggacag tgttggctct
     1681 ccgtgctccc gagtgggtga ggaggagcac gtctacagct tccccaacaa gcagaaatca
     1741 gctgagcctt cacccaccgt aatgagcacg tccctgggca gcaacctttc tgaactcgac
     1801 cgcctgctgc tggaactgaa cgctgtacag cataacccgc caggcttccc tgcagatgag
     1861 gccaactcaa gccccccgct tcctggggcc ctgagccccc tctatggtgt cccagagact
     1921 aacagcccct tgggaggcaa agctgggccc ctgacgaaag agaagcctaa gcggaatggg
     1981 ggccggggcc tggaggacgt gcggcccagt gtggagagtc tcttggatga actggagagc
     2041 tccgtgccca gccccgtccc tgccatcact gtgaaccagg gcgagatgag cagcccgcag
     2101 cgcgtcacct ccacccaaca gcagacacgc atctcggcct cctctgccac cagggagctg
     2161 gacgagctga tggcttcgct gtcggatttc aagttcatgg cccaggggaa gacagggagc
     2221 agctcacccc ctggggggcc cccgaagccc gggagccagc tggacagcat gctggggagc
     2281 ctgcagtctg acctgaacaa gctgggggtc gccacagtcg ccaaaggagt ctgcggggcc
     2341 tgcaagaagc ccatcgccgg gcaggttgtg accgccatgg ggaagacgtg gcaccccgag
     2401 cacttcgtct gcacccactg ccaggaggag atcggatccc ggaacttctt cgagcgggat
     2461 ggacagccct actgtgaaaa ggactaccac aacctcttct ccccgcgctg ctactactgc
     2521 aacggcccca tcctggataa agtggtgaca gcccttgacc ggacgtggca ccctgaacac
     2581 ttcttctgtg cacagtgtgg agccttcttt ggtcccgaag ggttccacga gaaggacggc
     2641 aaggcctact gtcgcaagga ctacttcgac atgttcgcac ccaagtgtgg cggctgcgcc
     2701 cgggccatcc tggagaacta tatctcagcc ctcaacacgc tgtggcatcc tgagtgcttt
     2761 gtgtgccggg aatgcttcac gccattcgtg aacggcagct tcttcgagca cgacgggcag
     2821 ccctactgtg aggtgcacta ccacgagcgg cgcggctcgc tgtgttctgg ctgccagaag
     2881 cccatcaccg gccgctgcat caccgccatg gccaagaagt tccaccccga gcacttcgtc
     2941 tgtgccttct gcctcaagca gctcaacaag ggcaccttca aggagcagaa cgacaagcct
     3001 tactgtcaga actgcttcct caagctcttc tgctagcgaa ttctgcagtc gacggtaccg
     3061 cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     3121 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3181 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3241 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3301 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3361 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3421 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3481 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3541 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3601 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3661 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3721 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3781 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3841 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3901 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3961 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4021 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4081 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4141 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4201 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4261 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4321 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4381 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4441 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4501 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4561 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4621 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4681 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4741 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4801 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4861 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4921 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4981 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5041 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5101 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5161 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5221 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5281 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5341 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5401 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5461 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5521 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5581 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5641 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5701 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5761 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5821 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     5881 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5941 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6001 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6061 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6121 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6181 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6241 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6301 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6361 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
