LOCUS       sequence-321638-        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(-9a)
FEATURES             Location/Qualifiers
     source          1..8627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     177..196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     430..447
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(521..625)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        735..1038
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1039..1242
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1193..1213
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1239..1263
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1368..1384
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1368..1384
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1395..1413
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1395..1412
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1414..1438
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5046..5067)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5307..5326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5654..5675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5765..5789)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5790..5809)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5791..5809)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5820..5837)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5820..5836)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5829..5851)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    6241..6375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6278..6297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6332..6351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6501..6955
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6588..6607)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6797..6818
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(7242..7310,7311..8102)
                     /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(7460..7479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    8204..8237
                     /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(8315..8627,1..276)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
       61 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      121 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      181 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      241 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      301 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      361 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
      421 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
      481 ctccccgcgc gttggccgat tcattaatgc agagcttgca attcgcgctt tttcaatatt
      541 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
      601 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
      661 aaaccattat tatcatgaca ttaacctata aaaataggcg tattacgagg ccctttcacg
      721 cattagatgc atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      781 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      841 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      901 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      961 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1021 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     1081 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     1141 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     1201 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
     1261 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
     1321 agcctccgga ctctagccta ggccgcggga cggataacaa tttcacacag gaaacagcta
     1381 tgaccattag gcctatttag gtgacactat agaacaagtt tgtacaaaaa agcaggctgg
     1441 taccggtccg gaattcccgg gatatcgtcg acccacgcgt ccgttaagtt ccgcgcgcag
     1501 gccaggctgc aacctgacgg ccagatccct cgctgtccta gtcgctgctc cttggagtca
     1561 tgttcccagc cgccccttct ccgcggaccc cgggtaccgg gtcccgaagg ggcccgctgg
     1621 ccggactcgg gcccggctcc acgccccgga cggctagcag gaagggtctg cccctggggt
     1681 ctgcagtcag ctccccagtg ctcttctcgc cggtcggccg gcgtagctcg ctaagctcgc
     1741 ggggaacacc aacacgaatg ttcccacacc actccataac tgagtctgtg aactatgatg
     1801 tgaaaacgtt tggatcttct cttcctgtta aagtcatgga agccctaaca ttggctgaag
     1861 tcgatgacca gctgaccatt aacatagatg aaggtggatg ggcttgtctg gtgtgcaaag
     1921 agaagctcat tatttggaag attgctctgt cacctattac taagttatcc gtttgcaaag
     1981 aacttcagct gccacctagt gatttccact ggagtgccga cttagtggct ctttcttact
     2041 cttctccctc aggtgaagca cattctactc aggctgttgc tgtcatggtt gccaccagag
     2101 aaggatctat ccgctattgg ccaagccttg ctggtgaaga tacctacaca gaggcttttg
     2161 tagattcggg aggtgataag acttacagtt tcctaacagc agtgcaggga ggaagtttta
     2221 ttttgtcttc atcaggaagc caactaattc ggttgatacc tgagagctca ggaaagattc
     2281 atcagcatat cctgcctcag gggcaaggca tgctttcagg aattggtcga aaagtttctt
     2341 ctctttttgg aattttatct cctagtagtg atctcacact ttcaagtgtt ctctgggata
     2401 gagagagatc aagcttttat agcctgacga gttcaaacat cagtaaatgg gaattagatg
     2461 attcttcaga aaagcatgca tacagttggg atataaatag agccctgaag gaaaacatta
     2521 ccgatgctat ttggggatct gaaagtaact atgaagctat taaagaagga gtcaacattc
     2581 gatatttgga ctttaagcaa aactgtgatg ggctggtgat tttggcagca gcatggcact
     2641 cagcagacaa tccatgtctc atctattact ctctgataac aatagaagat aatggttgcc
     2701 aaatgtcaga tgcagttact gtagaagtca ctcaatataa tccacctttt cagtctgaag
     2761 acctgatttt gtgtcagttg acggtcccaa acttttcaaa ccagactgcc tatctgtata
     2821 acgaaagtgc tgtctatgtg tgctccacag gaactgggaa attttctctt ccccaggaga
     2881 aaattgtctt taatgcacaa ggagatagtg ttttaggtgc tggtgcctgt ggtggtgttc
     2941 ctatcatttt ttctagaaac agtggactgg tgtctattac ttcaagggaa aatgtgtcta
     3001 tattggcaga agacttggaa gggtctttag catcttcagt tgctggacca aacagtgaga
     3061 gtatgatttt tgagaccact acaaagaatg aaactatagc ccaggaagat aaaatcaagt
     3121 tgctgaaagc tgcctttctg caatactgca gaaaagattt aggtcatgct caaatggtgg
     3181 ttgatgagct cttttcctct cactctgatt tggattctga ttctgaacta gacagggcag
     3241 ttacccaaat cagtgtagac ctgatggatg actacccagc atctgaccca cggtgggctg
     3301 agtctgtccc tgaggaagca cctgggttca gcaatacgtc actgattatc cttcaccagc
     3361 tagaagacaa gatgaaagct cactcttttc ttatggactt tattcatcaa gttggcttat
     3421 ttggacgtct aggcagtttt ccagttagag ggacaccgat ggccactcga ctgttgctct
     3481 gtgagcatgc cgaaaagctg tcagccgcca ttgttctcaa gaaccaccac tcccggcttt
     3541 ctgaccttgt caacacagcc atattgattg ctttgaacaa gagggagtat gaaatcccat
     3601 ccaacctgac tcctgcagat gtctttttca gggaggtatc ccaagtagat accatctgtg
     3661 agtgcttact ggagcatgag gagcaagtct tgagggatgc acctatggat tccattgaat
     3721 gggctgaagt ggtgatcaat gtgaacaata ttctcaagga tatgctgcag gctgctagtc
     3781 attatcgcca aaatagaaac tctttgtata gaagagaaga atcactagaa aaagaacctg
     3841 aatatgttcc atggacggca acaagtggtc ctggtggcat ccgaacggta ataatacgcc
     3901 agcatgagat tgtcctgaag gtggcttatc cacaggcaga cagcaacctc cgaaacatcg
     3961 tgaccgagca gctggtagcc ctgatcgatt gcttcctgga tggttatgtt tctcagctta
     4021 agtctgtgga taaatccagt aatcgggaaa gatatgacaa tctggagatg gaatacctac
     4081 agaaaagatc agatctctta tctcctcttc tttcactagg ccagtacctg tgggctgctt
     4141 ctctagcaga gaaatactgt gactttgata tattggtaca aatgtgtgag cagactgaca
     4201 accagagccg actccagcgc tacatgaccc agtttgctga tcagaatttt tcagactttc
     4261 tcttccgttg gtatctggag aaaggaaagc gaggcaaatt attatctcag cccatttctc
     4321 agcatggaca gttggcaaat tttttgcaag ctcatgaaca tctcagctgg ttacatgaaa
     4381 ttaatagcca agaattagaa aaggctcatg caacacttct gggtttggca aatatggaaa
     4441 ctcgttactt tgcaaagaag aaaacccttc ttggcttgag taaattggct gcattagctt
     4501 cagacttttc agaggatatg ctacaagaaa aaattgaaga aatggctgag caggagcgct
     4561 ttctactgca tcaggagacc ctacctgaac agctgctggc ggagaaacag ctaaatctca
     4621 gtgcgatgcc agtattgact gcaccacaac tcattggtct atatatctgt gaagaaaata
     4681 gaagagctaa tgaatatgat ttcaagaaag ctttggactt gttggaatat attgatgagg
     4741 aagaagatat aaatataaat gatctaaaac tggaaatcct ttgcaaagct cttcagagag
     4801 ataactggtc cagttctgat ggcaaagatg atccaattga agtatctaaa gacagtatat
     4861 ttgtgaagat cttacagaaa cttttaaaag atggcattca gctcagtgag tacttaccgg
     4921 aggtgaaaga cctgctacaa gcggatcagc ttggaagctt aaagtccaat ccttacttcg
     4981 agtttgtttt gaaagcaaat tatgaatatt atgttgagga gctgttcacc ggggtggtgc
     5041 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     5101 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     5161 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     5221 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     5281 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     5341 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     5401 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     5461 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     5521 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     5581 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     5641 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     5701 tggacgagct gtacaagtaa gcggccgctc tagagtatcc ctcgaggggc ccaagcttac
     5761 gcgtacccag ctttcttgta caaagtggtc cctatagtga gtcgtattat aagctaggca
     5821 ctggccgtcg ttttacaacg tcgtgactgg gaaaactgct agcttgggat ctttgtgaag
     5881 gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct
     5941 aaggtaaata taaaattttt aagtgtataa tgtgttaaac tagctgcata tgcttgctgc
     6001 ttgagagttt tgcttactga gtatgattta tgaaaatatt atacacagga gctagtgatt
     6061 ctaattgttt gtgtatttta gattcacagt cccaaggctc atttcaggcc cctcagtcct
     6121 cacagtctgt tcatgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     6181 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     6241 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6301 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6361 tatcatgtct ggatcgatcc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6421 gcgtattggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     6481 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6541 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6601 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6661 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6721 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6781 gtccacgttc ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6841 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6901 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     6961 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7021 atatgtatcc gctcatgcca ggtcttggac tggtgagaac ggcttgctcg gcagcttcga
     7081 tgtgtgctgg agggagaata aaggtctaag atgtgcgata gagggaagtc gcattgaatt
     7141 atgtgctgtg tagggatcgc tggtatcaaa tatgtgtgcc cacccctggc atgagacaat
     7201 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7261 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     7321 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     7381 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     7441 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     7501 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     7561 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     7621 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     7681 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     7741 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     7801 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     7861 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     7921 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7981 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     8041 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     8101 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     8161 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gccataactt cgtataatgt
     8221 atgctatacg aagttatggc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     8281 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8341 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8401 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8461 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8521 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8581 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcg
//
