LOCUS       sequence-321635-        8630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Nup133 fused at its carboxyl-terminus to 
            mEGFP with no linker and a total net fusion of (-8 amino acids) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8630)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 8630)
  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"
COMMENT     Sequence Label: Nup133_mEGFP(-8b)
FEATURES             Location/Qualifiers
     source          1..8630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        181..484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        485..688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     639..659
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     685..709
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     814..830
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     814..830
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        841..859
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     841..858
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    860..884
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4495..4516)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4756..4775)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5103..5124
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5214..5238)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5239..5258)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5240..5258)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5269..5286)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5269..5285)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5278..5300)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    5690..5824
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5727..5746)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5781..5800
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5950..6404
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6037..6056)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6246..6267
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(6691..6759,6760..7551)
                     /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     complement(6909..6928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    7653..7686
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      7764..8352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8253..8272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8506..8523
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(join(8597..8630,1..71))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
       61 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
      121 atgacattaa cctataaaaa taggcgtatt acgaggccct ttcacgcatt agatgcatgt
      181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      661 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      721 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      781 agcctaggcc gcgggacgga taacaatttc acacaggaaa cagctatgac cattaggcct
      841 atttaggtga cactatagaa caagtttgta caaaaaagca ggctggtacc ggtccggaat
      901 tcccgggata tcgtcgaccc acgcgtccgt taagttccgc gcgcaggcca ggctgcaacc
      961 tgacggccag atccctcgct gtcctagtcg ctgctccttg gagtcatgtt cccagccgcc
     1021 ccttctccgc ggaccccggg taccgggtcc cgaaggggcc cgctggccgg actcgggccc
     1081 ggctccacgc cccggacggc tagcaggaag ggtctgcccc tggggtctgc agtcagctcc
     1141 ccagtgctct tctcgccggt cggccggcgt agctcgctaa gctcgcgggg aacaccaaca
     1201 cgaatgttcc cacaccactc cataactgag tctgtgaact atgatgtgaa aacgtttgga
     1261 tcttctcttc ctgttaaagt catggaagcc ctaacattgg ctgaagtcga tgaccagctg
     1321 accattaaca tagatgaagg tggatgggct tgtctggtgt gcaaagagaa gctcattatt
     1381 tggaagattg ctctgtcacc tattactaag ttatccgttt gcaaagaact tcagctgcca
     1441 cctagtgatt tccactggag tgccgactta gtggctcttt cttactcttc tccctcaggt
     1501 gaagcacatt ctactcaggc tgttgctgtc atggttgcca ccagagaagg atctatccgc
     1561 tattggccaa gccttgctgg tgaagatacc tacacagagg cttttgtaga ttcgggaggt
     1621 gataagactt acagtttcct aacagcagtg cagggaggaa gttttatttt gtcttcatca
     1681 ggaagccaac taattcggtt gatacctgag agctcaggaa agattcatca gcatatcctg
     1741 cctcaggggc aaggcatgct ttcaggaatt ggtcgaaaag tttcttctct ttttggaatt
     1801 ttatctccta gtagtgatct cacactttca agtgttctct gggatagaga gagatcaagc
     1861 ttttatagcc tgacgagttc aaacatcagt aaatgggaat tagatgattc ttcagaaaag
     1921 catgcataca gttgggatat aaatagagcc ctgaaggaaa acattaccga tgctatttgg
     1981 ggatctgaaa gtaactatga agctattaaa gaaggagtca acattcgata tttggacttt
     2041 aagcaaaact gtgatgggct ggtgattttg gcagcagcat ggcactcagc agacaatcca
     2101 tgtctcatct attactctct gataacaata gaagataatg gttgccaaat gtcagatgca
     2161 gttactgtag aagtcactca atataatcca ccttttcagt ctgaagacct gattttgtgt
     2221 cagttgacgg tcccaaactt ttcaaaccag actgcctatc tgtataacga aagtgctgtc
     2281 tatgtgtgct ccacaggaac tgggaaattt tctcttcccc aggagaaaat tgtctttaat
     2341 gcacaaggag atagtgtttt aggtgctggt gcctgtggtg gtgttcctat cattttttct
     2401 agaaacagtg gactggtgtc tattacttca agggaaaatg tgtctatatt ggcagaagac
     2461 ttggaagggt ctttagcatc ttcagttgct ggaccaaaca gtgagagtat gatttttgag
     2521 accactacaa agaatgaaac tatagcccag gaagataaaa tcaagttgct gaaagctgcc
     2581 tttctgcaat actgcagaaa agatttaggt catgctcaaa tggtggttga tgagctcttt
     2641 tcctctcact ctgatttgga ttctgattct gaactagaca gggcagttac ccaaatcagt
     2701 gtagacctga tggatgacta cccagcatct gacccacggt gggctgagtc tgtccctgag
     2761 gaagcacctg ggttcagcaa tacgtcactg attatccttc accagctaga agacaagatg
     2821 aaagctcact cttttcttat ggactttatt catcaagttg gcttatttgg acgtctaggc
     2881 agttttccag ttagagggac accgatggcc actcgactgt tgctctgtga gcatgccgaa
     2941 aagctgtcag ccgccattgt tctcaagaac caccactccc ggctttctga ccttgtcaac
     3001 acagccatat tgattgcttt gaacaagagg gagtatgaaa tcccatccaa cctgactcct
     3061 gcagatgtct ttttcaggga ggtatcccaa gtagatacca tctgtgagtg cttactggag
     3121 catgaggagc aagtcttgag ggatgcacct atggattcca ttgaatgggc tgaagtggtg
     3181 atcaatgtga acaatattct caaggatatg ctgcaggctg ctagtcatta tcgccaaaat
     3241 agaaactctt tgtatagaag agaagaatca ctagaaaaag aacctgaata tgttccatgg
     3301 acggcaacaa gtggtcctgg tggcatccga acggtaataa tacgccagca tgagattgtc
     3361 ctgaaggtgg cttatccaca ggcagacagc aacctccgaa acatcgtgac cgagcagctg
     3421 gtagccctga tcgattgctt cctggatggt tatgtttctc agcttaagtc tgtggataaa
     3481 tccagtaatc gggaaagata tgacaatctg gagatggaat acctacagaa aagatcagat
     3541 ctcttatctc ctcttctttc actaggccag tacctgtggg ctgcttctct agcagagaaa
     3601 tactgtgact ttgatatatt ggtacaaatg tgtgagcaga ctgacaacca gagccgactc
     3661 cagcgctaca tgacccagtt tgctgatcag aatttttcag actttctctt ccgttggtat
     3721 ctggagaaag gaaagcgagg caaattatta tctcagccca tttctcagca tggacagttg
     3781 gcaaattttt tgcaagctca tgaacatctc agctggttac atgaaattaa tagccaagaa
     3841 ttagaaaagg ctcatgcaac acttctgggt ttggcaaata tggaaactcg ttactttgca
     3901 aagaagaaaa cccttcttgg cttgagtaaa ttggctgcat tagcttcaga cttttcagag
     3961 gatatgctac aagaaaaaat tgaagaaatg gctgagcagg agcgctttct actgcatcag
     4021 gagaccctac ctgaacagct gctggcggag aaacagctaa atctcagtgc gatgccagta
     4081 ttgactgcac cacaactcat tggtctatat atctgtgaag aaaatagaag agctaatgaa
     4141 tatgatttca agaaagcttt ggacttgttg gaatatattg atgaggaaga agatataaat
     4201 ataaatgatc taaaactgga aatcctttgc aaagctcttc agagagataa ctggtccagt
     4261 tctgatggca aagatgatcc aattgaagta tctaaagaca gtatatttgt gaagatctta
     4321 cagaaacttt taaaagatgg cattcagctc agtgagtact taccggaggt gaaagacctg
     4381 ctacaagcgg atcagcttgg aagcttaaag tccaatcctt acttcgagtt tgttttgaaa
     4441 gcaaattatg aatattatgt tcaggaggag ctgttcaccg gggtggtgcc catcctggtc
     4501 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4561 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4621 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     4681 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4741 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4801 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4861 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     4921 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     4981 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     5041 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     5101 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5161 tacaagtaag cggccgctct agagtatccc tcgaggggcc caagcttacg cgtacccagc
     5221 tttcttgtac aaagtggtcc ctatagtgag tcgtattata agctaggcac tggccgtcgt
     5281 tttacaacgt cgtgactggg aaaactgcta gcttgggatc tttgtgaagg aaccttactt
     5341 ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat
     5401 aaaattttta agtgtataat gtgttaaact agctgcatat gcttgctgct tgagagtttt
     5461 gcttactgag tatgatttat gaaaatatta tacacaggag ctagtgattc taattgtttg
     5521 tgtattttag attcacagtc ccaaggctca tttcaggccc ctcagtcctc acagtctgtt
     5581 catgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     5641 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     5701 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5761 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5821 gatcgatcct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattggct
     5881 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     5941 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6001 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6061 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6121 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6181 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6241 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6301 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6361 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     6421 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6481 ctcatgccag gtcttggact ggtgagaacg gcttgctcgg cagcttcgat gtgtgctgga
     6541 gggagaataa aggtctaaga tgtgcgatag agggaagtcg cattgaatta tgtgctgtgt
     6601 agggatcgct ggtatcaaat atgtgtgccc acccctggca tgagacaata accctgataa
     6661 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6721 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6781 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6841 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6901 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6961 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     7021 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     7081 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     7141 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7201 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     7261 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7321 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7381 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7441 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7501 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7561 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7621 taggtgaaga tcctttttga taatctcatg ccataacttc gtataatgta tgctatacga
     7681 agttatggca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7741 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7801 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7861 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7921 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7981 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     8041 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     8101 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     8161 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8221 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8281 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8341 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8401 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     8461 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     8521 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     8581 taatgcagag cttgcaattc gcgctttttc aatattattg aagcatttat
//
