LOCUS       sequence-321639-        8627 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 (-9 amino acids) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8627)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 8627)
  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(-9b)
FEATURES             Location/Qualifiers
     source          1..8627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     215..232
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(306..410)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        520..823
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        824..1027
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     978..998
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1024..1048
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1153..1169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1153..1169
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1180..1198
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1180..1197
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1199..1223
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4831..4852)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5092..5111)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5439..5460
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5550..5574)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5575..5594)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5576..5594)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5605..5622)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5605..5621)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5614..5636)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    6026..6160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6063..6082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6117..6136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6286..6740
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6373..6392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6582..6603
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(7027..7095,7096..7887)
                     /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(7245..7264)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    7989..8022
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      join(8100..8627,1..61)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8589..8608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
       61 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      121 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      181 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      241 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagagc
      301 ttgcaattcg cgctttttca atattattga agcatttatc agggttattg tctcatgagc
      361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      421 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
      481 aggcgtatta cgaggccctt tcacgcatta gatgcatgtc gttacataac ttacggtaaa
      541 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      601 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      661 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      721 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      781 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      841 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      901 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      961 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     1021 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     1081 ccatagaaga caccgggacc gatccagcct ccggactcta gcctaggccg cgggacggat
     1141 aacaatttca cacaggaaac agctatgacc attaggccta tttaggtgac actatagaac
     1201 aagtttgtac aaaaaagcag gctggtaccg gtccggaatt cccgggatat cgtcgaccca
     1261 cgcgtccgtt aagttccgcg cgcaggccag gctgcaacct gacggccaga tccctcgctg
     1321 tcctagtcgc tgctccttgg agtcatgttc ccagccgccc cttctccgcg gaccccgggt
     1381 accgggtccc gaaggggccc gctggccgga ctcgggcccg gctccacgcc ccggacggct
     1441 agcaggaagg gtctgcccct ggggtctgca gtcagctccc cagtgctctt ctcgccggtc
     1501 ggccggcgta gctcgctaag ctcgcgggga acaccaacac gaatgttccc acaccactcc
     1561 ataactgagt ctgtgaacta tgatgtgaaa acgtttggat cttctcttcc tgttaaagtc
     1621 atggaagccc taacattggc tgaagtcgat gaccagctga ccattaacat agatgaaggt
     1681 ggatgggctt gtctggtgtg caaagagaag ctcattattt ggaagattgc tctgtcacct
     1741 attactaagt tatccgtttg caaagaactt cagctgccac ctagtgattt ccactggagt
     1801 gccgacttag tggctctttc ttactcttct ccctcaggtg aagcacattc tactcaggct
     1861 gttgctgtca tggttgccac cagagaagga tctatccgct attggccaag ccttgctggt
     1921 gaagatacct acacagaggc ttttgtagat tcgggaggtg ataagactta cagtttccta
     1981 acagcagtgc agggaggaag ttttattttg tcttcatcag gaagccaact aattcggttg
     2041 atacctgaga gctcaggaaa gattcatcag catatcctgc ctcaggggca aggcatgctt
     2101 tcaggaattg gtcgaaaagt ttcttctctt tttggaattt tatctcctag tagtgatctc
     2161 acactttcaa gtgttctctg ggatagagag agatcaagct tttatagcct gacgagttca
     2221 aacatcagta aatgggaatt agatgattct tcagaaaagc atgcatacag ttgggatata
     2281 aatagagccc tgaaggaaaa cattaccgat gctatttggg gatctgaaag taactatgaa
     2341 gctattaaag aaggagtcaa cattcgatat ttggacttta agcaaaactg tgatgggctg
     2401 gtgattttgg cagcagcatg gcactcagca gacaatccat gtctcatcta ttactctctg
     2461 ataacaatag aagataatgg ttgccaaatg tcagatgcag ttactgtaga agtcactcaa
     2521 tataatccac cttttcagtc tgaagacctg attttgtgtc agttgacggt cccaaacttt
     2581 tcaaaccaga ctgcctatct gtataacgaa agtgctgtct atgtgtgctc cacaggaact
     2641 gggaaatttt ctcttcccca ggagaaaatt gtctttaatg cacaaggaga tagtgtttta
     2701 ggtgctggtg cctgtggtgg tgttcctatc attttttcta gaaacagtgg actggtgtct
     2761 attacttcaa gggaaaatgt gtctatattg gcagaagact tggaagggtc tttagcatct
     2821 tcagttgctg gaccaaacag tgagagtatg atttttgaga ccactacaaa gaatgaaact
     2881 atagcccagg aagataaaat caagttgctg aaagctgcct ttctgcaata ctgcagaaaa
     2941 gatttaggtc atgctcaaat ggtggttgat gagctctttt cctctcactc tgatttggat
     3001 tctgattctg aactagacag ggcagttacc caaatcagtg tagacctgat ggatgactac
     3061 ccagcatctg acccacggtg ggctgagtct gtccctgagg aagcacctgg gttcagcaat
     3121 acgtcactga ttatccttca ccagctagaa gacaagatga aagctcactc ttttcttatg
     3181 gactttattc atcaagttgg cttatttgga cgtctaggca gttttccagt tagagggaca
     3241 ccgatggcca ctcgactgtt gctctgtgag catgccgaaa agctgtcagc cgccattgtt
     3301 ctcaagaacc accactcccg gctttctgac cttgtcaaca cagccatatt gattgctttg
     3361 aacaagaggg agtatgaaat cccatccaac ctgactcctg cagatgtctt tttcagggag
     3421 gtatcccaag tagataccat ctgtgagtgc ttactggagc atgaggagca agtcttgagg
     3481 gatgcaccta tggattccat tgaatgggct gaagtggtga tcaatgtgaa caatattctc
     3541 aaggatatgc tgcaggctgc tagtcattat cgccaaaata gaaactcttt gtatagaaga
     3601 gaagaatcac tagaaaaaga acctgaatat gttccatgga cggcaacaag tggtcctggt
     3661 ggcatccgaa cggtaataat acgccagcat gagattgtcc tgaaggtggc ttatccacag
     3721 gcagacagca acctccgaaa catcgtgacc gagcagctgg tagccctgat cgattgcttc
     3781 ctggatggtt atgtttctca gcttaagtct gtggataaat ccagtaatcg ggaaagatat
     3841 gacaatctgg agatggaata cctacagaaa agatcagatc tcttatctcc tcttctttca
     3901 ctaggccagt acctgtgggc tgcttctcta gcagagaaat actgtgactt tgatatattg
     3961 gtacaaatgt gtgagcagac tgacaaccag agccgactcc agcgctacat gacccagttt
     4021 gctgatcaga atttttcaga ctttctcttc cgttggtatc tggagaaagg aaagcgaggc
     4081 aaattattat ctcagcccat ttctcagcat ggacagttgg caaatttttt gcaagctcat
     4141 gaacatctca gctggttaca tgaaattaat agccaagaat tagaaaaggc tcatgcaaca
     4201 cttctgggtt tggcaaatat ggaaactcgt tactttgcaa agaagaaaac ccttcttggc
     4261 ttgagtaaat tggctgcatt agcttcagac ttttcagagg atatgctaca agaaaaaatt
     4321 gaagaaatgg ctgagcagga gcgctttcta ctgcatcagg agaccctacc tgaacagctg
     4381 ctggcggaga aacagctaaa tctcagtgcg atgccagtat tgactgcacc acaactcatt
     4441 ggtctatata tctgtgaaga aaatagaaga gctaatgaat atgatttcaa gaaagctttg
     4501 gacttgttgg aatatattga tgaggaagaa gatataaata taaatgatct aaaactggaa
     4561 atcctttgca aagctcttca gagagataac tggtccagtt ctgatggcaa agatgatcca
     4621 attgaagtat ctaaagacag tatatttgtg aagatcttac agaaactttt aaaagatggc
     4681 attcagctca gtgagtactt accggaggtg aaagacctgc tacaagcgga tcagcttgga
     4741 agcttaaagt ccaatcctta cttcgagttt gttttgaaag caaattatga atattatgtt
     4801 caggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     4861 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     4921 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     4981 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     5041 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     5101 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     5161 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     5221 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     5281 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     5341 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     5401 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     5461 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagcggc cgctctagag
     5521 tatccctcga ggggcccaag cttacgcgta cccagctttc ttgtacaaag tggtccctat
     5581 agtgagtcgt attataagct aggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5641 ctgctagctt gggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa
     5701 actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt
     5761 taaactagct gcatatgctt gctgcttgag agttttgctt actgagtatg atttatgaaa
     5821 atattataca caggagctag tgattctaat tgtttgtgta ttttagattc acagtcccaa
     5881 ggctcatttc aggcccctca gtcctcacag tctgttcatg atcataatca gccataccac
     5941 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     6001 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     6061 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6121 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gatcctgcat taatgaatcg
     6181 gccaacgcgc ggggagaggc ggtttgcgta ttggctggcg taatagcgaa gaggcccgca
     6241 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     6301 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6361 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6421 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     6481 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     6541 cggtttttcg ccctttgacg ttggagtcca cgttcttaat agtggactct tgttccaaac
     6601 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6661 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6721 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6781 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgccaggtct tggactggtg
     6841 agaacggctt gctcggcagc ttcgatgtgt gctggaggga gaataaaggt ctaagatgtg
     6901 cgatagaggg aagtcgcatt gaattatgtg ctgtgtaggg atcgctggta tcaaatatgt
     6961 gtgcccaccc ctggcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7021 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7081 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7141 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7201 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7261 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7321 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7381 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7441 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7501 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7561 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7621 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7681 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7741 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7801 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7861 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7921 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7981 ctcatgccat aacttcgtat aatgtatgct atacgaagtt atggcatgac caaaatccct
     8041 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8101 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8161 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     8221 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     8281 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8341 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8401 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8461 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8521 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8581 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcg
//
