LOCUS       sequence-220837-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mEGFP-linker sequence for N-terminus tagging of 
            human NUP153.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5418)
  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            "Unpublished"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-58: NUP153-mEGFP
FEATURES             Location/Qualifiers
     source          1..5418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..59
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     253..275
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     267..284
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     268..284
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1302..1304,1305..1307,1308..2018)
                     /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(1347..1368)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1608..1627)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1955..1976
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3086..3102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3086..3102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3099..3121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3110..3126
                     /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(3134..3140,3141..3158,3159..3164))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3179..3200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3317..3334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3488..4076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3568..3587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4247..5038,5039..5107))
                     /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     4870..4889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5108..5212)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5280..5298
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5336..5358)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
       61 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      121 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      181 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      241 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg acgcgtattg
      301 ggataacctt tatatgttta aagaaacatt aatgaagccg tatatatttt ctaattctaa
      361 ggatcatgga ctttattttt caaccacaga tttttttttc ctccttctta aatcattagc
      421 tcttgtcttt gattttactg tctgcttact tcctgcctac ttaaccaacc gacagggaga
      481 aggagaatgg gaagaggtta ctgtttttat gttaagtgtc ttttcaaagt ctaatttaca
      541 gccaagaaca gcaaaagtac cccttggctt tgtgcctctg cggcgccccg gagatctcct
      601 tccccgtccc tcttcctttc cgagaagccg cgttccgttg ttgaactacg tttcccagcg
      661 tgccgcgcgg ccgagagcgt ggactcgacc agcccttcct ctttgcctat cagctcgtcg
      721 cagcatgatg atgcaaatgc gtcgggcgcc gatgagtgac gacattctgg ctcgcgaccg
      781 cggcagccgc cttagcaggg gtaataggcg caacggcggc ggaggctgca gggacgacga
      841 cgacgacggc ggcggggcgg gcgctgtgcg cacaggggag gggtaggcgg gcgtcgccag
      901 cggcttcaac tgcggcagct acggctaggg cgacggaaga actcccacca gtcggcggtc
      961 gagttaggcc tcagcaccgc ggggaactgt tcgtgctgtc ctctgcggga gatctccata
     1021 gagaccgtga cacacacaga ggcgggggtc cgcagaggcg gccggaccgg agcctcctcg
     1081 gcctctgcgg ccgccacccc cttccccgct gccagcactc accctctctc cgctcctgct
     1141 cgcaacctcg cggttcctgg ggtgcttgtg cccactgtgt ggacagcgcg ggcggacttt
     1201 tgggccgggg ccgggcggcg ggggaggctc tctaaggcct ccgcctctgc ctctcccgcc
     1261 cccttacccg ccccggagcg ggaagcggcg gaggctccgc catggtgagc aagggcgagg
     1321 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1381 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1441 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1501 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1561 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1621 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1681 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1741 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1801 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1861 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     1921 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1981 ccgccgggat cactctcggc atggacgagc tgtacaagta cagcgacctg gaggcctcag
     2041 gggcaggagg agtcggaggg ggcggtggcg gcaagatccg gacgcggcgt tgccaccagg
     2101 ggccaattaa gccttaccag caggggcgac aacagcatca ggtatgggac cccgacgccg
     2161 cggtggcctg gcgggtgggg aaactcggct ggaggggacg ggcggttcga ggccccggcg
     2221 ccccacgtgg agcagacagg agggcctgag gaaaggcccg agtgaggctc cgggggagct
     2281 caggccgttg ggggtggcga cgggaagtgg gctgagacag gcccagcctt gcggcgcaca
     2341 cgtgtggttt gggccgcctt tcccccaact ctctgggtca ggggctttcc caggcaggct
     2401 ctgggcggga aaagaaattg taggcccatg atgcgcgcgt ttctttattt tttggcgggg
     2461 atttggaatt tgagggtatt actcatgttc tctcgaggga gaaaggaaaa gatgtggttg
     2521 atggccctta tacttgctgt tttctggtgt cccttcttta gtgtgtttga ggaggggggg
     2581 ctgccccttg gaatcttaac tctggggcag acggggagtt gtggggggcg ctgttacatg
     2641 gaaagtggaa taacttagtc tcagcctatg agttaattct cggtttcccc tgcctgtggc
     2701 cctcctcgtc ttgagagcat tggcttagat tttattaacc aggttatagt taaccgtggc
     2761 actggggcat tttgaaagtc cctctccacc cttacgtctt ttctctgctg cgcgactccc
     2821 ctcccccccc ccaacaccgc cacccccgcc tttattttgg aggctggaat tggccattct
     2881 ccacagttga ccgggactcc agctctgatc tcatacacaa aatattttaa tattatgagg
     2941 cgtattctag aagtatagtt acttttgaca ctccaggaat gttggtggtc ttatagttta
     3001 gccactgtat gaactcctgg atgtttaaaa ttaatcccaa tggcgcgccg aatcccaatg
     3061 gcgcgccgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3121 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3181 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3241 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3301 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3361 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3421 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3481 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3661 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3841 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3901 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3961 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4021 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4081 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4141 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4201 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4261 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4321 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4381 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4441 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4501 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4561 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4621 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4681 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4741 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4801 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4861 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4921 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4981 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5041 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5101 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5161 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5221 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5281 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     5341 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5401 aagcccgtca gggcgcgt
//
