LOCUS       sequence-321553-        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(-8a)
FEATURES             Location/Qualifiers
     source          1..8630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(79..183)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        293..596
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        597..800
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     751..771
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     797..821
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     926..942
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     926..942
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        953..971
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     953..970
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    972..996
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4607..4628)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4868..4887)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5215..5236
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5326..5350)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5351..5370)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5352..5370)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5381..5398)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5381..5397)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5390..5412)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    5802..5936
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5839..5858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5893..5912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6062..6516
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6149..6168)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6358..6379
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(6803..6871,6872..7663)
                     /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(7021..7040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    7765..7798
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      7876..8464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8365..8384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(8618..8630,1..5)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
       61 attaatgcag agcttgcaat tcgcgctttt tcaatattat tgaagcattt atcagggtta
      121 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      181 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
      241 aacctataaa aataggcgta ttacgaggcc ctttcacgca ttagatgcat gtcgttacat
      301 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      361 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      421 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      481 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      541 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      601 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      661 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      721 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      781 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      841 gctgttttga cctccataga agacaccggg accgatccag cctccggact ctagcctagg
      901 ccgcgggacg gataacaatt tcacacagga aacagctatg accattaggc ctatttaggt
      961 gacactatag aacaagtttg tacaaaaaag caggctggta ccggtccgga attcccggga
     1021 tatcgtcgac ccacgcgtcc gttaagttcc gcgcgcaggc caggctgcaa cctgacggcc
     1081 agatccctcg ctgtcctagt cgctgctcct tggagtcatg ttcccagccg ccccttctcc
     1141 gcggaccccg ggtaccgggt cccgaagggg cccgctggcc ggactcgggc ccggctccac
     1201 gccccggacg gctagcagga agggtctgcc cctggggtct gcagtcagct ccccagtgct
     1261 cttctcgccg gtcggccggc gtagctcgct aagctcgcgg ggaacaccaa cacgaatgtt
     1321 cccacaccac tccataactg agtctgtgaa ctatgatgtg aaaacgtttg gatcttctct
     1381 tcctgttaaa gtcatggaag ccctaacatt ggctgaagtc gatgaccagc tgaccattaa
     1441 catagatgaa ggtggatggg cttgtctggt gtgcaaagag aagctcatta tttggaagat
     1501 tgctctgtca cctattacta agttatccgt ttgcaaagaa cttcagctgc cacctagtga
     1561 tttccactgg agtgccgact tagtggctct ttcttactct tctccctcag gtgaagcaca
     1621 ttctactcag gctgttgctg tcatggttgc caccagagaa ggatctatcc gctattggcc
     1681 aagccttgct ggtgaagata cctacacaga ggcttttgta gattcgggag gtgataagac
     1741 ttacagtttc ctaacagcag tgcagggagg aagttttatt ttgtcttcat caggaagcca
     1801 actaattcgg ttgatacctg agagctcagg aaagattcat cagcatatcc tgcctcaggg
     1861 gcaaggcatg ctttcaggaa ttggtcgaaa agtttcttct ctttttggaa ttttatctcc
     1921 tagtagtgat ctcacacttt caagtgttct ctgggataga gagagatcaa gcttttatag
     1981 cctgacgagt tcaaacatca gtaaatggga attagatgat tcttcagaaa agcatgcata
     2041 cagttgggat ataaatagag ccctgaagga aaacattacc gatgctattt ggggatctga
     2101 aagtaactat gaagctatta aagaaggagt caacattcga tatttggact ttaagcaaaa
     2161 ctgtgatggg ctggtgattt tggcagcagc atggcactca gcagacaatc catgtctcat
     2221 ctattactct ctgataacaa tagaagataa tggttgccaa atgtcagatg cagttactgt
     2281 agaagtcact caatataatc caccttttca gtctgaagac ctgattttgt gtcagttgac
     2341 ggtcccaaac ttttcaaacc agactgccta tctgtataac gaaagtgctg tctatgtgtg
     2401 ctccacagga actgggaaat tttctcttcc ccaggagaaa attgtcttta atgcacaagg
     2461 agatagtgtt ttaggtgctg gtgcctgtgg tggtgttcct atcatttttt ctagaaacag
     2521 tggactggtg tctattactt caagggaaaa tgtgtctata ttggcagaag acttggaagg
     2581 gtctttagca tcttcagttg ctggaccaaa cagtgagagt atgatttttg agaccactac
     2641 aaagaatgaa actatagccc aggaagataa aatcaagttg ctgaaagctg cctttctgca
     2701 atactgcaga aaagatttag gtcatgctca aatggtggtt gatgagctct tttcctctca
     2761 ctctgatttg gattctgatt ctgaactaga cagggcagtt acccaaatca gtgtagacct
     2821 gatggatgac tacccagcat ctgacccacg gtgggctgag tctgtccctg aggaagcacc
     2881 tgggttcagc aatacgtcac tgattatcct tcaccagcta gaagacaaga tgaaagctca
     2941 ctcttttctt atggacttta ttcatcaagt tggcttattt ggacgtctag gcagttttcc
     3001 agttagaggg acaccgatgg ccactcgact gttgctctgt gagcatgccg aaaagctgtc
     3061 agccgccatt gttctcaaga accaccactc ccggctttct gaccttgtca acacagccat
     3121 attgattgct ttgaacaaga gggagtatga aatcccatcc aacctgactc ctgcagatgt
     3181 ctttttcagg gaggtatccc aagtagatac catctgtgag tgcttactgg agcatgagga
     3241 gcaagtcttg agggatgcac ctatggattc cattgaatgg gctgaagtgg tgatcaatgt
     3301 gaacaatatt ctcaaggata tgctgcaggc tgctagtcat tatcgccaaa atagaaactc
     3361 tttgtataga agagaagaat cactagaaaa agaacctgaa tatgttccat ggacggcaac
     3421 aagtggtcct ggtggcatcc gaacggtaat aatacgccag catgagattg tcctgaaggt
     3481 ggcttatcca caggcagaca gcaacctccg aaacatcgtg accgagcagc tggtagccct
     3541 gatcgattgc ttcctggatg gttatgtttc tcagcttaag tctgtggata aatccagtaa
     3601 tcgggaaaga tatgacaatc tggagatgga atacctacag aaaagatcag atctcttatc
     3661 tcctcttctt tcactaggcc agtacctgtg ggctgcttct ctagcagaga aatactgtga
     3721 ctttgatata ttggtacaaa tgtgtgagca gactgacaac cagagccgac tccagcgcta
     3781 catgacccag tttgctgatc agaatttttc agactttctc ttccgttggt atctggagaa
     3841 aggaaagcga ggcaaattat tatctcagcc catttctcag catggacagt tggcaaattt
     3901 tttgcaagct catgaacatc tcagctggtt acatgaaatt aatagccaag aattagaaaa
     3961 ggctcatgca acacttctgg gtttggcaaa tatggaaact cgttactttg caaagaagaa
     4021 aacccttctt ggcttgagta aattggctgc attagcttca gacttttcag aggatatgct
     4081 acaagaaaaa attgaagaaa tggctgagca ggagcgcttt ctactgcatc aggagaccct
     4141 acctgaacag ctgctggcgg agaaacagct aaatctcagt gcgatgccag tattgactgc
     4201 accacaactc attggtctat atatctgtga agaaaataga agagctaatg aatatgattt
     4261 caagaaagct ttggacttgt tggaatatat tgatgaggaa gaagatataa atataaatga
     4321 tctaaaactg gaaatccttt gcaaagctct tcagagagat aactggtcca gttctgatgg
     4381 caaagatgat ccaattgaag tatctaaaga cagtatattt gtgaagatct tacagaaact
     4441 tttaaaagat ggcattcagc tcagtgagta cttaccggag gtgaaagacc tgctacaagc
     4501 ggatcagctt ggaagcttaa agtccaatcc ttacttcgag tttgttttga aagcaaatta
     4561 tgaatattat gttcagggag agctgttcac cggggtggtg cccatcctgg tcgagctgga
     4621 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     4681 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     4741 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     4801 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     4861 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4921 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4981 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     5041 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     5101 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     5161 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     5221 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     5281 agcggccgct ctagagtatc cctcgagggg cccaagctta cgcgtaccca gctttcttgt
     5341 acaaagtggt ccctatagtg agtcgtatta taagctaggc actggccgtc gttttacaac
     5401 gtcgtgactg ggaaaactgc tagcttggga tctttgtgaa ggaaccttac ttctgtggtg
     5461 tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt
     5521 taagtgtata atgtgttaaa ctagctgcat atgcttgctg cttgagagtt ttgcttactg
     5581 agtatgattt atgaaaatat tatacacagg agctagtgat tctaattgtt tgtgtatttt
     5641 agattcacag tcccaaggct catttcaggc ccctcagtcc tcacagtctg ttcatgatca
     5701 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     5761 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     5821 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5881 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcgatc
     5941 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg ctggcgtaat
     6001 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6061 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6121 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6181 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6241 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6301 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
     6361 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     6421 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6481 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     6541 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgcc
     6601 aggtcttgga ctggtgagaa cggcttgctc ggcagcttcg atgtgtgctg gagggagaat
     6661 aaaggtctaa gatgtgcgat agagggaagt cgcattgaat tatgtgctgt gtagggatcg
     6721 ctggtatcaa atatgtgtgc ccacccctgg catgagacaa taaccctgat aaatgcttca
     6781 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6841 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6901 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6961 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7021 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7081 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7141 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7201 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7261 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7321 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7381 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7441 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7501 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7561 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7621 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7681 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7741 gatccttttt gataatctca tgccataact tcgtataatg tatgctatac gaagttatgg
     7801 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7861 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7921 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7981 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     8041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     8101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     8161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     8221 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     8281 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8341 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8401 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8461 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     8521 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     8581 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
//
