LOCUS       sequence-178082-        6871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mAPPLE labeled fluorescent actin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6871)
  AUTHORS   Sun J, Lu F, He H, Shen J, Messina J, Mathew R, Wang D, Sarnaik AA, 
            Chang WC, Kim M, Cheng H, Yang S
  TITLE     STIM1- and Orai1-mediated Ca(2+) oscillation orchestrates 
            invadopodium formation and melanoma invasion.
  JOURNAL   J Cell Biol. 2014 Nov 24;207(4):535-48. doi: 10.1083/jcb.201407082. 
            Epub 2014 Nov 17.
  PUBMED    25404747
REFERENCE   2  (bases 1 to 6871)
  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"; doi: 
            "10.1083/jcb.201407082"; journalName: "J Cell Biol"; date: 
            "2014-11-24- 24"; volume: "207"; issue: "4"; pages: "535-48"
COMMENT     Sequence Label: pLNCX2-lifeact-mAPPLE
FEATURES             Location/Qualifiers
     source          1..6871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             559..609
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             616..1326
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="VSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEAF
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNFE
                     DGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALKS
                     EIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERAE
                     GRHSTGGMDELYKS"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             1397..1990
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2093..2115)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2201..2341
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2256..2275
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2356..2373)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2527..3115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2607..2626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3286..4077,4078..4146))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3909..3928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4147..4251)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4319..4337
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             4909..5084
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5147..5346
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5547..5963
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5893..5915
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5931..5947
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             6007..6801
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6061..6080)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6671..6690
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagat gggcgtggcc gacctgatca agaagttcga gagcatcagc
      601 aaggaggagg gatccgtgag caagggcgag gagaataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgaggccttt cagaccgcta agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggtc
      841 tacattaagc acccagccga catccccgac tacttcaagc tgtccttccc cgagggcttc
      901 aggtgggagc gcgtgatgaa cttcgaggac ggcggcatta ttcacgttaa ccaggactcc
      961 tccctgcagg acggcgtgtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccgagga gcggatgtac
     1081 cccgaggacg gcgccctgaa gagcgagatc aagaagaggc tgaagctgaa ggacggcggc
     1141 cactacgccg ccgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacatcg tcgacatcaa gttggacatc gtgtcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tcctaagtcg acctttcctt aatggcctaa catcgataaa ataaaagatt ttatttagtc
     1381 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     1441 cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt
     1501 caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     1561 gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg
     1621 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca
     1681 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     1741 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     1801 ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga
     1861 ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     1921 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     1981 gtctttcatt tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca
     2041 ccaccgggag gtaagctggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2101 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2161 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     2221 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     2281 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     2341 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2701 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2941 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3241 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3301 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3361 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3421 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3481 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3541 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3601 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3661 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3721 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3781 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3841 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3901 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3961 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4021 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4081 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4141 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4201 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4261 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4321 taggcgtatc acgaggccct ttcgtcttca agaattcata ccagatcacc gaaaactgtc
     4381 ctccaaatgt gtccccctca cactcccaaa ttcgcgggct tctgcctctt agaccactct
     4441 accctattcc ccacactcac cggagccaaa gccgcggccc ttccgtttct ttgcttttga
     4501 aagaccccac ccgtaggtgg caagctagct taagtaacgc cactttgcaa ggcatggaaa
     4561 aatacataac tgagaataga aaagttcaga tcaaggtcag gaacaaagaa acagctgaat
     4621 accaaacagg atatctgtgg taagcggttc ctgccccggc tcagggccaa gaacagatga
     4681 gacagctgag tgatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcgg
     4741 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gtgaatcatc
     4801 agatgtttcc agggtgcccc aaggacctga aaatgaccct gtaccttatt tgaactaacc
     4861 aatcagttcg cttctcgctt ctgttcgcgc gcttccgctc tccgagctca ataaaagagc
     4921 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta
     4981 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg
     5041 gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg
     5101 atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
     5161 cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt
     5221 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
     5281 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
     5341 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     5401 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     5461 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     5521 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     5581 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     5641 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     5701 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     5761 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     5821 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     5881 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     5941 ccctcactcc ttctctaggc gccggaattc cgatctgatc aagagacagg atgaggatcg
     6001 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6061 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6121 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6181 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6241 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6301 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6361 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6421 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6481 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6541 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6601 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6661 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6721 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6781 cttcttgacg agttcttctg agcgggactc tggggttcga atagtaatca attacggggt
     6841 cattagttca tagcccatat atggagttcc g
//
