LOCUS       sequence-385098-        5957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene Overexpression of Human Arp3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5957)
  AUTHORS   Butt U, Khan MH, Pouwels J, Westermarck J
  TITLE     SHARPIN S146 phosphorylation mediates ARP2/3 interaction, cancer 
            cell invasion and metastasis.
  JOURNAL   J Cell Sci. 2022 Oct 15;135(20):jcs260627. doi: 10.1242/jcs.260627. 
            Epub 2022 Oct 13.
  PUBMED    36148554
REFERENCE   2  (bases 1 to 5957)
  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: "J Cell 
            Sci."; date: "2022-10-15"; volume: "135(20):jcs260627. doi"; pages: 
            " 10.1242/jcs.260627. Epub 2022 Oct 13"
COMMENT     Sequence Label: Arp3-TagRFP
FEATURES             Location/Qualifiers
     source          1..5957
                     /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      1904..1913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1910..2623
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    2745..2866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2782..2801)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2836..2855
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2873..3328)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3010..3031)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3222..3241
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3355..3459
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3461..3818
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3461..3652
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3484..3504)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3669..3804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3853..4647
                     /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             3853..4647
                     /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(3907..3926)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4517..4536
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4835..4854)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4879..4926
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5255..5843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5744..5763
                     /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 caagcttcga attcatggcg ggacggctgc cggcctgtgt
      661 ggtggactgt ggcacggggt atacaaaact aggatatgct ggaaatacag aaccacagtt
      721 tatcatccct tcctgtattg ctattaagga gtcagcaaaa gtgggtgatc aagctcaaag
      781 gagggtgatg aaaggtgttg atgacctaga cttcttcatt ggtgatgaag caatagaaaa
      841 acctacatat gcaacaaagt ggccaatccg ccatggtata gttgaagatt gggacttaat
      901 ggaaaggttt atggagcaag tgatctttaa atatttaagg gcagaacctg aagaccatta
      961 ttttcttttg actgaacctc cattgaatac tccagaaaac agggaatata ctgctgaaat
     1021 aatgtttgag tccttcaatg ttccaggctt gtacattgct gtgcaggctg ttcttgcctt
     1081 agctgcatct tggacctcaa gacaagtagg agaacggacg ttgaccggta cggtaataga
     1141 cagtggagat ggtgtcactc atgtcattcc tgtggctgaa gggtatgtga ttggcagctg
     1201 tattaaacac attccaatcg caggacgaga tataacatat tttattcagc aactgctgag
     1261 agaccgagaa gtaggaatcc ctccagaaca atccttggaa actgctaagg cagtaaagga
     1321 gcgctatagt tatgtctgcc cagatttagt aaaagaattt aacaagtatg atacagatgg
     1381 gtcaaaatgg attaaacagt atactggaat caatgctatc tcaaagaaag agttttctat
     1441 cgatgttggt tatgagagat ttttgggacc tgaaatcttt tttcatccag agtttgctaa
     1501 tccagacttt acacaaccta tctcagaagt tgtagatgaa gtaattcaga attgtcctat
     1561 tgatgtcaga cgtcctctct acaagaatat tgtcctctct ggaggttcaa ccatgttcag
     1621 ggactttgga cgtcgcttgc aaagagattt gaaaagaact gtagatgccc ggctgaaatt
     1681 aagtgaggaa ttgagtggtg gtagattgaa gccaaaacct attgatgtac aagtcattac
     1741 acaccacatg cagcgatatg cagtttggtt tggaggatca atgctggctt ccacgcctga
     1801 gttctaccaa gtatgccaca ccaaaaagga ttatgaagaa attggaccta gcatttgtcg
     1861 tcacaatcca gtgtttggag tcatgtcgac ggatccaccg gtcgccacca tggtgtctaa
     1921 gggcgaagag ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgaa
     1981 caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac
     2041 catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac
     2101 cagcttcatg tacggcagca gaaccttcat caaccacacc cagggcatcc ccgacttctt
     2161 taagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg
     2221 cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa
     2281 gatcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa cactcggctg
     2341 ggaggccaac accgagatgc tgtaccccgc tgacggcggc ctggaaggca gaagcgacat
     2401 ggccctgaag ctcgtgggcg ggggccacct gatctgcaac ttcaagacca catacagatc
     2461 caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tatgtggacc acagactgga
     2521 aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg ctgtggccag
     2581 atactgcgac ctccctagca aactggggca caaacttaat tgagcggccg cgactctaga
     2641 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2701 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2761 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2821 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     2881 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2941 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3001 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3061 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3121 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3181 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3241 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3301 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3361 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3421 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3481 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3541 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3601 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3661 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3721 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3781 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     3841 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3901 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3961 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4021 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4081 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4141 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4201 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4261 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4321 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4381 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4441 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4501 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4561 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4621 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4681 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4741 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4801 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     4861 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     4921 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4981 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5041 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5101 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5161 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5221 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5281 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5341 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5401 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5461 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5521 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5581 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5641 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5701 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5761 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5821 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5881 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5941 gtattaccgc catgcat
//
