LOCUS       sequence-290199-        6291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNot1iso1_1085-1605-del3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6291)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6291)
  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: pT7-EGFP-C1-HsNot1iso1_1085-1605-del3_V
FEATURES             Location/Qualifiers
     source          1..6291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3079..3200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3116..3135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3170..3189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3207..3662)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3344..3365)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3556..3575
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3689..3793
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3795..4152
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3795..3986
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3818..3838)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4003..4138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4187..4981
                     /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             4187..4981
                     /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(4241..4260)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4851..4870
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5169..5188)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5213..5260
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5589..6177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6078..6097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctactgagag aattgtggag cccccagaaa atatccaggg
     1441 tagtagtggt atgacacaaa aggttgaaga gctaaaggaa acggtgaaag aagaatttat
     1501 gccttgggtt tcacagtatc tggttatgaa gagagtcagt attgagccaa actttcatag
     1561 cctgtattca aacttccttg acacgctgaa gaatcctgaa tttaacaaga tggttctgaa
     1621 tgagacctac agaaacatta aagtgctcct gacctctgat aaagctgcag ccaatttctc
     1681 agatcgttct ttgctgaaga acttgggaca ttggctagga atgatcacat tagctaaaaa
     1741 caaacccatc ttacacactg acttggatgt gaaatcattg ctgctagagg cttatgttaa
     1801 aggacaacaa gaattgctct atgtagtgcc ctttgttgcc aaagtcttag aatctagcat
     1861 tcgtagtgtg gtttttaggc caccaaaccc ttggacaatg gcaattatga atgtattagc
     1921 tgagctacat caggagcatg acttaaagtt aaacttgaag tttgaaatcg aggttctctg
     1981 caagaacctt gcattagaca tcaatgagct aaaacctgga aacctcctaa aggataaaga
     2041 tcgcctgaag aatttagatg agcaactctc tgctccaaag aaagatgtca agcagccaga
     2101 agaactccct cccatcacaa ccacaacaac ttctactaca ccagctacca acaccacttg
     2161 tacagccacg gttccaccac agccacagta cagctaccac gacatcaatg tctattccct
     2221 tgcgggcttg gcaccacaca ttactctaaa tccaacaatt cccttgtttc aggcccatcc
     2281 acagttgaag cagtgtgtgc gtcaggcaat tgaacgggct gtccaggagc tggtccatcc
     2341 tgtggtggat cgatcaatta agattgccat gactacttgt gagcaaatag tcaggaagga
     2401 ttttgccctg gattcggagg aatctcgaat gcgaatagca gctcatcaca tgatgcgtaa
     2461 cttgacagct ggaatggcta tgattacatg cagggaacct ttgctcatga gcatatctac
     2521 caacttaaaa aacagttttg cctcagccct tcgtactgct tccccacaac aaagagaaat
     2581 gatggatcag gcagctgctc aattagctca ggacaattgt gagttggctt gctgttttat
     2641 tcagaagact gcagtagaaa aagcaggccc tgagatggac aagagattag caactgaatt
     2701 tgagctgaga aaacatgcta ggcaagaagg acgcagatac tgtgatcctg ttgttttaac
     2761 atatcaagct gaacggatgc cagagcaaat caggctgaaa gttggtggtg tggacccaaa
     2821 gcagttggct gtttacgaag agtttgcacg caatgttcct ggcttcttac ctacaaatga
     2881 cttaagtcag cccacgggat ttttagccca gcccatgaag caagcttggg caacatgacc
     2941 gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     3001 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3061 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3121 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3181 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3241 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3301 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3361 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3421 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3481 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3541 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3601 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3661 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3721 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3781 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3901 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3961 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4021 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4081 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4141 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4201 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4261 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4321 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4381 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4441 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4501 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4561 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4621 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4681 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4741 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4801 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4861 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4921 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4981 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5041 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5101 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5161 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5221 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5281 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5341 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5401 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5461 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5521 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5581 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5641 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5701 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5761 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5821 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5881 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5941 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6001 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6061 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6181 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6241 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
