LOCUS       sequence-316723-        7682 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of Nup93 fused at its carboxyl-terminus to 
            mEGFP with no linker and a total net fusion of (-6 amino acids) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7682)
  TITLE     Simon lab plasmids
REFERENCE   2  (bases 1 to 7682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Nup93_mEGFP(-6)
FEATURES             Location/Qualifiers
     source          1..7682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(284..303)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    328..375
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      704..1292
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1193..1212
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1467..1770
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1771..1974
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1925..1945
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(4566..4587)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4827..4846)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5174..5195
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5876..5997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5913..5932)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5967..5986
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6004..6459)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6141..6162)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6353..6372
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6486..6590
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6592..6949
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6592..6783
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6615..6635)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6800..6935
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(6984..7682,1..96)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(6984..7682,1..96)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7038..7057)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7648..7667
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
       61 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
      121 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
      181 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
      241 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
      301 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
      361 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
      421 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
      481 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
      541 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
      601 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
      661 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
      721 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
      781 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
      841 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
      901 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
      961 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     1021 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     1081 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     1141 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     1201 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     1261 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     1321 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     1381 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
     1441 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     1501 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1561 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1621 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1681 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1741 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     1801 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     1861 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     1921 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
     1981 gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat gtccggactc agatctcgac
     2041 ttcccgggat gacggctgta gcacaaggat ctgcatctcc aatggatact gaggggtttg
     2101 gtgagctcct tcagcaagct gaacagcttg ctgctgagac tgagggcatc tcagagcttc
     2161 cccatgtgga acggaactta caggagatcc agcaggcggg agagcgcctg cgttcccgta
     2221 ccctaacacg cacgtcccag gagacggcag atgtcaaggc gtcagttctc ctcgggtctc
     2281 ggggacttga catatcccac atctcccagc gattggagag tctgagtgca gccaccacct
     2341 ttgagcctct tgagcctgtg aaggacactg acattcaggg cttcctgaag aatgagaagg
     2401 acaatgccct gctgtctgcc atcgaagagt cccggaagag gaccttcggc atggctgagg
     2461 agtaccatcg ggagtcaatg ttggttgagt gggagcaagt gaaacagcga attctgcaca
     2521 cactgctggc atcaggagaa gacgcccttg actttactca agaaagcgag ccaagctaca
     2581 tcagtgatgt gggaccccct ggtcgaagct ctctggataa catcgagatg gcctatgcgc
     2641 ggcaaattta tatctataat gagaaaattg taaatggaca cctgcagcct aacctggtgg
     2701 acctttgtgc ttccgtcgca gagctcgatg ataagagcat ttccgacatg tggaccatgg
     2761 taaaacaaat gacagacgtg ttgttgacac cggcaacgga tgccctgaag aaccgcagca
     2821 gcgtggaagt gcgcatggag tttgtcaggc aggccttggc gtaccttgag cagagttata
     2881 agaattacac ccttgtgact gtctttggaa atttgcatca ggcccagctg ggcggggtgc
     2941 ctgggactta ccaattggtt cgaagtttcc tgaacattaa actgccagct cccttgcctg
     3001 gactacagga tggagaggtg gaaggccatc ctgtgtgggc gctaatttac tactgcatgc
     3061 gctgtggaga cctgcttgcc gcttcacagg tagttaatcg agcccagcac cagctgggag
     3121 agtttaaaac ctggttccag gagtacatga acagcaagga cagaagattg tccccagcta
     3181 cggaaaacaa gctccggctg cattaccgta gggccctcag gaacaataca gatccctaca
     3241 agcgggccgt gtactgtatc attggcagat gtgacgtcac cgacaaccag agtgaagtgg
     3301 cggacaaaac tgaggattac ctgtggctga agttgaacca agtgtgtttt gacgacgatg
     3361 gcaccagctc cccacaagac aggctcactc tctcacagtt ccagaagcag ttgttggaag
     3421 actatggcga gtcccacttt acggtgaacc agcaaccctt cctctacttc caagtcctgt
     3481 tcctgacagc gcagtttgaa gcagcagttg cctttctttt ccgcatggag cggctgcgct
     3541 gccatgctgt ccatgtagca ctggtgctgt ttgagctgaa gctgctttta aagtcctctg
     3601 gacagagggc tcagctcctc agccacgagc ctggtgaccc tccttgcttg cggcggctga
     3661 acttcgtgcg gctcctcatg ctgtacaccc ggaagtttga gtccacggac ccaagggagg
     3721 ccctccagta cttctatttc ctcagggatg agaaagatag tcaaggagaa aacatgtttc
     3781 tgcgctgtgt gagtgagctt gtgattgaaa gccgagagtt cgatatgatt cttgggaaac
     3841 tagagaatga cggaagtaga aagcctggag tcatagataa gtttactagt gacacaaagc
     3901 ctattatcaa caaagttgct tctgtggcag aaaataaagg actgtttgaa gaggcagcaa
     3961 agctgtatga ccttgccaag aatgctgaca aggtactgga gctgatgaac aaactgctga
     4021 gccctgtcgt cccccagatc agtgccccgc aatccaacaa ggagaggctg aagaacatgg
     4081 cactctccat tgccgaacgg tatagggctc aaggaataag cgcaaataaa tttgtggact
     4141 ccacgttcta tcttcttttg gacttgatca ccttttttga cgagtatcat agtggtcata
     4201 ttgatagagc ttttgatatc attgagcgct tgaagctggt gcccctgaat caggaaagtg
     4261 tggaagagag agtggctgcc ttcagaaatt tcagtgatga aatcaggcac aacctctcag
     4321 aagtgcttct tgccaccatg aacatcttgt tcacacagtt taagaggctc aaggggacaa
     4381 gtccatcctc gtcatccagg ccccagcgag tcatcgagga ccgcgactct caactccgaa
     4441 gtcaagcccg cactctgatt acctttgctg gaatgatacc ataccgaacg tctggggaca
     4501 ccaatgcgag gctggtgcag atggaggtcc tcatgaatga gctgttcacc ggggtggtgc
     4561 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     4621 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     4681 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     4741 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     4801 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     4861 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     4921 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     4981 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     5041 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     5101 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     5161 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     5221 tggacgagct gtacaagtaa gtgccatgct ttgtgggagt ctgggtcggc acactgtcag
     5281 tacatcaggc acatgggccc actaggctgg ggtttctggt tttgtttctg ttgtgttttg
     5341 ttttggtttc tgtattatgt atttttgtca acgccaataa atttctttga tttggtgaat
     5401 cgcatcgagc tgaccggcac cgatttcaag gaggatggca acatcctggg caataagatg
     5461 gagtacaact acaacgccca caatgtgtac atcatgaccg acaaggccaa gaatggcatc
     5521 aaggtgaact tcaagatccg ccacaacatc gaggatggca gcgtgcagct ggccgaccac
     5581 taccagcaga atacccccat cggcgatggc cctgtgctgc tgcccgataa ccactacctg
     5641 tccacccaga gcgccctgtc caaggacccc aacgagaagc gcgatcacat gatctacttc
     5701 ggcttcgtga cctctagagt atccctcgag gatccgagct ccgcggagat ccaccggatc
     5761 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     5821 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     5881 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5941 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     6001 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6061 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6121 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6181 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6241 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6301 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6361 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6421 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     6481 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6541 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     6601 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     6661 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     6721 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     6781 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6841 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6901 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     6961 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     7021 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     7081 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     7141 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     7201 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     7261 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     7321 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     7381 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     7441 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     7501 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     7561 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     7621 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     7681 gc
//
