LOCUS       sequence-289564-        6451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNxf-1-delLRR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6451)
  AUTHORS   Teplova M, Wohlbold L, Khin NW, Izaurralde E, Patel DJ
  TITLE     Structure-function studies of nucleocytoplasmic transport of 
            retroviral genomic RNA by mRNA export factor TAP.
  JOURNAL   Nat Struct Mol Biol. 2011 Aug 7;18(9):990-8. doi: 10.1038/nsmb.2094.
  PUBMED    21822283
REFERENCE   2  (bases 1 to 6451)
  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"; doi: "10.1038/nsmb"; 
            journalName: "Nat Struct Mol Biol"; date: "2011-08-7- 7"; volume: 
            "18"; issue: "9"; pages: "990-8"
COMMENT     Sequence Label: pT7-EGFP-C1-HsNXF-1delLRR_O
FEATURES             Location/Qualifiers
     source          1..6451
                     /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    3239..3360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3276..3295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3330..3349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3367..3822)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3504..3525)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3716..3735
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3849..3953
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3955..4312
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3955..4146
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3978..3998)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4163..4298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4347..5141
                     /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             4347..5141
                     /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(4401..4420)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5011..5030
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5329..5348)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5373..5420
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5749..6337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6238..6257
                     /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 ctcaagcttc gaattctgca gtcgccatgg atatggcgga
     1441 cgaggggaag tcgtacagcg aacacgatga tgaacgcgtt aatttccctc aaagaaagaa
     1501 gaaaggccgg ggtcccttcc ggtggaaata tggtgaagga aaccgtaggt ctggaagagg
     1561 cggttctggt attcggtctt cccgccttga ggaagatgat ggagatgtgg caatgagtga
     1621 tgcccaggat ggtccccgag tacgatacaa cccctatacc acccgaccta accgtcgggg
     1681 tgatacttgg catgatcgag atcgcattca tgttactgtg cggagagaca gagctcctcc
     1741 agagagagga ggggctggca ccagccagga tgggacctca aagaactggt tcaagattac
     1801 aattccttat ggcagaaagt atgacaaggc atggctcctg agcatgattc agagcaagtg
     1861 cagtgtgccc ttcaccccta ttgagtttca ctatgagaat acacgggccc agttcttcgt
     1921 tgaagacgcc agtactgcct ctgcattgaa ggctgtcaac tataagattt tggatcggga
     1981 gaaccgaagg atatctatca tcatcaactc ttctgctcca ccccacacta tactgaatat
     2041 ccctgagcta ttgtccttga acttgagcaa caacaggctg tacaggctgg atgacatgtc
     2101 tagcattgtt cagaaggcac ccaacctgaa gatcctaaac ctttctggaa atgaattgaa
     2161 gtctgagcgg gaattggaca agataaaggg gctgaagcta gaagagctct ggctcgatgg
     2221 aaactccctg tgtgacacct tccgagacca gtccacctac atcagcgcca ttcgcgaacg
     2281 atttcccaag ttactacgcc tggatggcca tgagctaccc ccaccaattg cctttgatgt
     2341 tgaagccccc acgacgttac cgccctgcaa gggaagctat tttggaacag aaaacttgaa
     2401 gagtctggtc ttgcacttcc tgcaacagta ctatgcaatt tacgactctg gagaccgaca
     2461 agggctcctg gatgcctacc atgatggggc ctgctgttcc ctgagcattc ctttcattcc
     2521 tcagaaccct gcccgaagca gcttagccga gtatttcaag gatagcagaa atgtgaagaa
     2581 gcttaaagac cctaccttgc ggttccggct gctgaagcac acgcgtctca acgttgttgc
     2641 cttcctcaat gagttgccca aaacccagca cgacgtcaat tccttcgtgg tagacataag
     2701 cgcccagaca agcacattgc tgtgtttttc tgtcaatgga gtcttcaagg aagtggacgg
     2761 aaagtcccgg gattctttgc gagccttcac ccggacattc attgctgttc ctgctagcaa
     2821 ttcagggcta tgtattgtaa atgatgagct atttgtgcgg aatgccagtt ctgaagagat
     2881 ccaaagagcc ttcgctatgc ctgcacccac gccttcctcc agcccggtgc ccaccctctc
     2941 tccagagcag caggaaatgt tgcaagcatt ctctacccag tctggcatga acctcgagtg
     3001 gtcccagaag tgccttcagg acaacaactg ggactacacc agatctgccc aggccttcac
     3061 tcatctcaag gccaagggcg agatcccaga agtggcattc atgaagtgag gatccaccgg
     3121 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3181 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3241 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3301 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3361 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3421 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3481 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3541 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3601 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3661 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3721 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3781 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3841 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3901 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     3961 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4021 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4081 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4141 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4201 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4261 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4321 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4381 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4441 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4501 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4561 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4621 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4681 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4741 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4801 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4861 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4921 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4981 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5041 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5101 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5161 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5221 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5281 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5341 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5401 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5461 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5521 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5581 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5641 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5701 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5761 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5821 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5881 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5941 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6001 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6061 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6121 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6181 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6241 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6301 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6361 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6421 ttctgtggat aaccgtatta ccgccatgca t
//
