LOCUS       sequence-220813-        5429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mTagRFPT-linker sequence for N-terminus tagging of
            human LMNB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5429)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5429)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-29: LMNB1-mTagRFP-T
FEATURES             Location/Qualifiers
     source          1..5429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..30
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     22..39
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     23..39
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1059..1790
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     primer_bind     complement(2852..2868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2852..2868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2865..2887)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2876..2892
                     /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(2900..2906,2907..2924,2925..2930))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2945..2966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3083..3100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3254..3842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3334..3353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4013..4804,4805..4873))
                     /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     4636..4655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4874..4978)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5046..5064
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5102..5124)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5224..5243
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgacgcg tattgggatt
       61 aaaggctggt acttggaacc tgcaagccgt gcatttggaa cctcggactc aagtgcctat
      121 tacgtaattc cacagcgtcc cggcctccag gccgtttccc gagccctcca gcggagcggg
      181 ggataaggtt accacgcccg cggtggccgg ggacactctg agtttcgcgt gtggctttta
      241 gggacgttta tatttgaatt tccctgaacc gccgagtgtg ggcggtggcg cagatccgtc
      301 ccggaaacct ccgggctcct tcccgccttt ctcaggcccg gcccctccaa ggggtccccg
      361 cggggcggcg ggagggccct gggcccagag ccgcgcgggt gggcagtccc aggcgtcctt
      421 ccttacagcc ctgagcctgg tccgggaacc gcccagccgg gagggccgag ctgacggttg
      481 cccaagggcc agattttaaa tttacaggcc cggcccccga accgccgaag cgcgctgcct
      541 gctccccatt ggcccatggt agtcacgtgg aggcgccggg gcgtgccggc catgttgggg
      601 agtgcggcgc cgcggcccgc gccacctccg ccccccgcgg cttgcctcca gcccgcccct
      661 cccggccctc ctccccccgc ccgccgctcc gtgcagcctg agaggaaaca aagtgctgcg
      721 agcaggagac ggcggcggcg cgaaccctgc tgggcctcca gtcaccctcg tcttgcattt
      781 tcccgcgtgc gtgtgtgagt gggtgtgtgt gttttcttac aaagggtatt tcgcgatcga
      841 tcgattgatt cgtagttccc ccccgcgcgc ctttgccctt tgtgctgtaa tcgagctccc
      901 gccatcccag gtgcttctcc gttcctctaa acgccagcgt ctggacgtga gcgcaggtcg
      961 ccggtttgtg ccttcggtcc ccgcttcgcc ccctgccgtc ccctccttat cacggtcccg
     1021 ctcgcggcct cgccgccccg ctgtctccgc cgcccgccat ggtgtctaag ggcgaagagc
     1081 tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact
     1141 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca
     1201 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt
     1261 acggcagcag aaccttcatc aaccacaccc agggcatccc cgatttcttt aagcagtcct
     1321 tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg
     1381 ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg
     1441 tgaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccaaca
     1501 ccgagatgct gtaccccgct gacggcggcc tggaaggcag aaccgacatg gccctgaagc
     1561 tcgtgggcgg gggccacctg atctgcaact tcaagaccac atacagatcc aagaaacccg
     1621 ctaagaacct caagatgccc ggcgtctact atgtggacca cagactggaa agaatcaagg
     1681 aggccgacaa agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc
     1741 tccctagcaa actggggcac aaacttaatg gcatggacga gctgtacaag agcggcctga
     1801 gaagcagagc ccaggccagc gcgactgcga cccccgtgcc gccgcggatg ggcagccgcg
     1861 ctggcggccc caccacgccg ctgagcccca cgcgcctgtc gcggctccag gagaaggagg
     1921 agctgcgcga gctcaatgac cggctggcgg tgtacatcga caaggtgcgc agcctggaga
     1981 cggagaacag cgcgctgcag ctgcaggtga cggagcgcga ggaggtgcgc ggccgtgagc
     2041 tcaccggcct caaggcgctc tacgagaccg agctggccga cgcgcgacgc gcgctcgacg
     2101 acacggcccg cgagcgcgcc aagctgcaga tcgagctggg caagtgcaag gcggaacacg
     2161 accagctgct cctcaagtga gtgctagctg gcggccgcgt tagcgccaag gaggggcggg
     2221 ggcgcaaccg cggcgaccag ctcaccgggt tctgccgtgg ggagggagca gaggccagga
     2281 tgcacgcgtc cttctgaagg aacagggtct cggtctccgg aaaggagaaa gaatctagag
     2341 ttcatagcgg agcaggggtc gcggaggggg ctcgagctgt agcgctgggg ggccgtgatg
     2401 cccatttcta gattttggat acccgctggg acgtggtaag tgcgcgcctg ggactgccga
     2461 gaaggagctc ccgctttcgc actcgaatcc ggggagccgg cgcggagagg cggcccctca
     2521 ggccccaggt gcggggagct ggagcgcgag cgcgcgctcg cgtgcgcgcc ccagtttccg
     2581 gccggcgcga gacaaagcgt ctagcggatt tgcagtgccg ggatgggcgg ccggggagga
     2641 ctggcagccc gcctctagaa tgaatgagct tcgcgcgggc agagagagga aggggaggga
     2701 ccttcccgca gcatccgcgt ctcctggggg tgggtcccgc tttggcgcgc tcagtcttgg
     2761 ccctgtgacg ttttgcgaag attctacgcc tgctttaggc gggagagaga ggcggagatc
     2821 ccaatggcgc gccgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2881 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2941 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3001 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3061 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3121 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3661 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3901 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3961 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4021 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4081 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4141 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4201 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4261 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4321 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4381 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4441 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4501 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4561 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4621 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4681 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4741 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4801 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4861 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4921 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4981 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5041 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     5101 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     5161 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     5221 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     5281 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     5341 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     5401 ctgcaaggcg attaagttgg gtaacgcca
//
