LOCUS       sequence-173949-        5414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and linker-mEGFP sequence for N-terminus tagging of 
            human LMNB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5414)
  AUTHORS   Roberts B, Haupt A, Tucker A, Grancharova T, Arakaki J, Fuqua MA, 
            Nelson A, Hookway C, Ludmann SA, Mueller IA, Yang R, Horwitz R, 
            Rafelski SM, Gunawardane RN
  TITLE     Systematic gene tagging using CRISPR/Cas9 in human stem cells to 
            illuminate cell organization.
  JOURNAL   Mol Biol Cell. 2017 Oct 15;28(21):2854-2874. doi: 
            10.1091/mbc.E17-03-0209. Epub 2017 Aug 16.
  PUBMED    28814507
REFERENCE   2  (bases 1 to 5414)
  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: 
            "10.1091/mbc.E17-03-0209"; journalName: "Mol Biol Cell"; date: 
            "2017-10-15- 15"; volume: "28"; issue: "21"; pages: "2854-2874"
COMMENT     Sequence Label: AICSDP-10:LMNB1-mEGFP
FEATURES             Location/Qualifiers
     source          1..5414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1415..1417,1418..1420,1421..2131)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1460..1481)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1721..1740)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2068..2089
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3193..3209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3193..3209)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3206..3228)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3217..3233
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3241..3247,3248..3265,3266..3271))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3286..3307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3424..3441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3595..4183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3675..3694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4354..5145,5146..5214))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4977..4996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5215..5319)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5387..5405
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggattaaag
      421 gctggtactt ggaacctgca agccgtgcat ttggaacctc ggactcaagt gcctattacg
      481 taattccaca gcgtcccggc ctccaggccg tttcccgagc cctccagcgg agcgggggat
      541 aaggttacca cgcccgcggt ggccggggac actctgagtt tcgcgtgtgg cttttaggga
      601 cgtttatatt tgaatttccc tgaaccgccg agtgtgggcg gtggcgcaga tccgtcccgg
      661 aaacctccgg gctccttccc gcctttctca ggcccggccc ctccaagggg tccccgcggg
      721 gcggcgggag ggccctgggc ccagagccgc gcgggtgggc agtcccaggc gtccttcctt
      781 acagccctga gcctggtccg ggaaccgccc agccgggagg gccgagctga cggttgccca
      841 agggccagat tttaaattta caggcccggc ccccgaaccg ccgaagcgcg ctgcctgctc
      901 cccattggcc catggtagtc acgtggaggc gccggggcgt gccggccatg ttggggagtg
      961 cggcgccgcg gcccgcgcca cctccgcccc ccgcggcttg cctccagccc gcccctcccg
     1021 gccctcctcc ccccgcccgc cgctccgtgc agcctgagag gaaacaaagt gctgcgagca
     1081 ggagacggcg gcggcgcgaa ccctgctggg cctccagtca ccctcgtctt gcattttccc
     1141 gcgtgcgtgt gtgagtgggt gtgtgtgttt tcttacaaag ggtatttcgc gatcgatcga
     1201 ttgattcgta gttccccccc gcgcgccttt gccctttgtg ctgtaatcga gctcccgcca
     1261 tcccaggtgc ttctccgttc ctctaaacgc cagcgtctgg acgtgagcgc aggtcgccgg
     1321 tttgtgcctt cggtccccgc ttcgccccct gccgtcccct ccttatcacg gtcccgctcg
     1381 cggcctcgcc gccccgctgt ctccgccgcc cgccatggtg agcaagggcg aggagctgtt
     1441 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1501 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1561 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1621 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1681 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1741 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1801 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1861 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1921 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1981 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     2041 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2101 gatcactctc ggcatggacg agctgtacaa gagcggcctg agaagcagag cccaggccag
     2161 cgcgactgcg acccccgtgc cgccgcggat gggcagccgc gctggcggcc ccaccacgcc
     2221 gctgagcccc acgcgcctgt cgcggctcca ggagaaggag gagctgcgcg agctcaatga
     2281 ccggctggcg gtgtacatcg acaaggtgcg cagcctggag acggagaaca gcgcgctgca
     2341 gctgcaggtg acggagcgcg aggaggtgcg cggccgtgag ctcaccggcc tcaaggcgct
     2401 ctacgagacc gagctggccg acgcgcgacg cgcgctcgac gacacggccc gcgagcgcgc
     2461 caagctgcag atcgagctgg gcaagtgcaa ggcggaacac gaccagctgc tcctcaagtg
     2521 agtgctagct ggcggccgcg ttagcgccaa ggaggggcgg gggcgcaacc gcggcgacca
     2581 gctcaccggg ttctgccgtg gggagggagc agaggccagg atgcacgcgt ccttctgaag
     2641 gaacagggtc tcggtctccg gaaaggagaa agaatctaga gttcatagcg gagcaggggt
     2701 cgcggagggg gctcgagctg tagcgctggg gggccgtgat gcccatttct agattttgga
     2761 tacccgctgg gacgtggtaa gtgcgcgcct gggactgccg agaaggagct cccgctttcg
     2821 cactcgaatc cggggagccg gcgcggagag gcggcccctc aggccccagg tgcggggagc
     2881 tggagcgcga gcgcgcgctc gcgtgcgcgc cccagtttcc ggccggcgcg agacaaagcg
     2941 tctagcggat ttgcagtgcc gggatgggcg gccggggagg actggcagcc cgcctctaga
     3001 atgaatgagc ttcgcgcggg cagagagagg aaggggaggg accttcccgc agcatccgcg
     3061 tctcctgggg gtgggtcccg ctttggcgcg ctcagtcttg gccctgtgac gttttgcgaa
     3121 gattctacgc ctgctttagg cgggagagag aggcggagat cccaatggcg cgccgagctt
     3181 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3241 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3301 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3361 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3421 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3481 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3541 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3601 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3661 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3721 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3781 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3841 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3901 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3961 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4021 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4081 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4141 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4201 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4261 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4321 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4381 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4441 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4501 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4561 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4621 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4681 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4741 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4801 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4861 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4921 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4981 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5041 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5101 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5161 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5221 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5281 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5341 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5401 gaggcccttt cgtc
//
