LOCUS       sequence-260227-        6373 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 6373)
  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 6373)
  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..6373
                     /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    3161..3282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3198..3217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3252..3271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3289..3744)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3426..3447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3638..3657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3771..3875
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3877..4234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3877..4068
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3900..3920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4085..4220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4269..5063
                     /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="MIEQDGLHEGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTESGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4323..4342)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4933..4952
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5251..5270)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5295..5342
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5671..6259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6160..6179
                     /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 tgctaggctc gagtctagat aactgatcat
     3061 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3121 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3181 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3241 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3301 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3361 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3421 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3481 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3541 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3601 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3661 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3721 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3781 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3841 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3901 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4081 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4141 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4201 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4261 cgtttcgcat gattgaacaa gatggattgc acgaaggttc tccggccgct tgggtggaga
     4321 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4381 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4441 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4501 cagctgtgct cgacgttgtc actgaatcgg gaagggactg gctgctattg ggcgaagtgc
     4561 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4621 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4681 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4741 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4801 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4861 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4921 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4981 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5041 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5101 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5161 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5221 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5281 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5341 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5401 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5461 aggcccaggg ctcgcagcca acgtcggggc ggcaggcact gccatagcct caggttactc
     5521 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5581 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5641 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5701 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5761 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5821 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5881 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5941 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6001 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6061 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6121 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6181 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6241 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6301 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6361 taccgccatg cat
//
