LOCUS       sequence-405389-        6754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-tagged Lamin A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  AUTHORS   Warner H, Franciosa G, van der Borg G, Coenen B, Faas F, Koenig C, 
            de Boer R, Classens R, Maassen S, Baranov MV, Mahajan S, Dabral D, 
            Bianchi F, van Hilten N, Risselada HJ, Roos WH, Olsen JV, Cano LQ, 
            van den Bogaart G
  TITLE     Atypical cofilin signaling drives dendritic cell migration through 
            the extracellular matrix via nuclear deformation.
  JOURNAL   Cell Rep. 2024 Feb 27;43(3):113866. doi: 
            10.1016/j.celrep.2024.113866.
  PUBMED    38416638
REFERENCE   2  (bases 1 to 6754)
  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: "Cell Rep.";
            date: "2024-02-27"; volume: "43(3):113866. doi"; pages: " 
            10.1016/j.celrep.2024.113866"
COMMENT     Sequence Label: GFP-Lamin-A
FEATURES             Location/Qualifiers
     source          1..6754
                     /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             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3542..3663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3579..3598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3633..3652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3670..4125)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3807..3828)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4019..4038
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4152..4256
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4258..4615
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4258..4449
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4281..4301)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4466..4601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4650..5444
                     /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             4650..5444
                     /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(4704..4723)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5314..5333
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5632..5651)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5676..5723
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6052..6640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6541..6560
                     /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 gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgaggc ggcggcggcg gcggcggcgg cggcggcggc
     1381 ggcggcggcg aaaccccgag ccagcgccgc gcgacccgca gcggcgcgca ggcgagcagc
     1441 accccgctga gcccgacccg cattacccgc ctgcaggaaa aagaagatct gcaggaactg
     1501 aacgatcgcc tggcggtgta tattgatcgc gtgcgcagcc tggaaaccga aaacgcgggc
     1561 ctgcgcctgc gcattaccga aagcgaagaa gtggtgagcc gcgaagtgag cggcattaaa
     1621 gcggcgtatg aagcggaact gggcgatgcg cgcaaaaccc tggatagcgt ggcgaaagaa
     1681 cgcgcgcgcc tgcagctgga actgagcaaa gtgcgcgaag aatttaaaga actgaaagcg
     1741 cgcaacacca aaaaagaagg cgatctgatt gcggcgcagg cgcgcctgaa agatctggaa
     1801 gcgctgctga acagcaaaga agcggcgctg agcaccgcgc tgagcgaaaa acgcaccctg
     1861 gaaggcgaac tgcatgatct gcgcggccag gtggcgaaac tggaagcggc gctgggcgaa
     1921 gcgaaaaaac agctgcagga tgaaatgctg cgccgcgtgg atgcggaaaa ccgcctgcag
     1981 accatgaaag aagaactgga ttttcagaaa aacatttata gcgaagaact gcgcgaaacc
     2041 aaacgccgcc atgaaacccg cctggtggaa attgataacg gcaaacagcg cgaatttgaa
     2101 agccgcctgg cggatgcgct gcaggaactg cgcgcgcagc atgaagatca ggtggaacag
     2161 tataaaaaag aactggaaaa aacctatagc gcgaaactgg ataacgcgcg ccagagcgcg
     2221 gaacgcaaca gcaacctggt gggcgcggcg catgaagaac tgcagcagag ccgcattcgc
     2281 attgatagcc tgagcgcgca gctgagccag ctgcagaaac agctggcggc gaaagaagcg
     2341 aaactgcgcg atctggaaga tagcctggcg cgcgaacgcg ataccagccg ccgcctgctg
     2401 gcggaaaaag aacgcgaaat ggcggaaatg cgcgcgcgca tgcagcagca gctggatgaa
     2461 tatcaggaac tgctggatat taaactggcg ctggatatgg aaattcatgc gtatcgcaaa
     2521 ctgctggaag gcgaagaaga acgcctgcgc ctgagcccga gcccgaccag ccagcgcagc
     2581 cgcggccgcg cgagcagcca tagcagccag acccagggcg gcggcagcgt gaccaaaaaa
     2641 cgcaaactgg aaagcaccga aagccgcagc agctttagcc agcatgcgcg caccagcggc
     2701 cgcgtggcgg tggaagaagt ggatgaagaa ggcaaatttg tgcgcctgcg caacaaaagc
     2761 aacgaagatc agagcatggg caactggcag attaaacgcc agaacggcga tgatccgctg
     2821 ctgacctatc gctttccgcc gaaatttacc ctgaaagcgg gccaggtggt gaccatttgg
     2881 gcggcgggcg cgggcgcgac ccatagcccg ccgaccgatc tggtgtggaa agcgcagaac
     2941 acctggggct gcggcaacag cctgcgcacc gcgctgatta acagcaccgg cgaagaagtg
     3001 gcgatgcgca aactggtgcg cagcgtgacc gtggtggaag atgatgaaga tgaagatggc
     3061 gatgatctgc tgcatcatca tcatggcagc cattgcagca gcagcggcga tccggcggaa
     3121 tataacctgc gcagccgcac cgtgctgtgc ggcacctgcg gccagccggc ggataaagcg
     3181 agcgcgagcg gcagcggcgc gcaggtgggc ggcccgatta gcagcggcag cagcgcgagc
     3241 agcgtgaccg tgacccgcag ctatcgcagc gtgggcggca gcggcggcgg cagctttggc
     3301 gataacctgg tgacccgcag ctatctgctg ggcaacagca gcccgcgcac ccagagcccg
     3361 cagaactgca gcattatgta agaattctgc agtcgacggt accgcgggcc cgggatccac
     3421 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3481 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3541 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3601 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3661 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3721 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3781 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3841 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3901 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3961 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4021 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4081 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     4141 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4201 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     4261 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4321 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4381 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4441 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4501 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4561 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     4621 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4681 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4741 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4801 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4861 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4921 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4981 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5041 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5101 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5161 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5221 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5281 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5341 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5401 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5461 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5521 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5581 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     5641 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5701 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5761 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5821 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5881 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5941 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6001 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6061 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6121 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6181 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6241 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6301 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6361 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6421 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6481 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6541 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6601 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6661 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6721 tgattctgtg gataaccgta ttaccgccat gcat
//
