LOCUS       sequence-356535-        6011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HseIF4A3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  AUTHORS   Palacios IM, Gatfield D, St Johnston D, Izaurralde E
  TITLE     An eIF4AIII-containing complex required for mRNA localization and 
            nonsense-mediated mRNA decay.
  JOURNAL   Nature. 2004 Feb 19. 427(6976):753-7.
  PUBMED    14973490
REFERENCE   2  (bases 1 to 6011)
  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"; journalName: "Nature. 
            2004 Feb 19. 427(6976):753-7."
COMMENT     Sequence Label: pT7-EGFP-C1-HseIF4A3_I
FEATURES             Location/Qualifiers
     source          1..6011
                     /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    2799..2920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2836..2855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2890..2909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2927..3382)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3064..3085)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3276..3295
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3409..3513
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3515..3872
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3515..3706
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3538..3558)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3723..3858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3907..4701
                     /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             3907..4701
                     /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(3961..3980)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4571..4590
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4889..4908)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4933..4980
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5309..5897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5798..5817
                     /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 ctatggcgac cacggccacg atggcgacct cgggctcggc
     1441 gcgaaagcgg ctgctcaaag aggaagacat gactaaagtg gaattcgaga ccagcgagga
     1501 ggtggatgtg acccccacgt tcgacaccat gggcctgcgg gaggacctgc tgcggggcat
     1561 ctacgcttac ggttttgaaa aaccatcagc aatccagcaa cgagcaatca agcagatcat
     1621 caaagggaga gatgtcatcg cacagtctca gtccggcaca ggaaaaacag ccaccttcag
     1681 tatctcagtc ctccagtgtt tggatattca ggttcgtgaa actcaagctt tgatcttggc
     1741 tcccacaaga gagttggctg tgcagatcca gaaggggctg cttgctctcg gtgactacat
     1801 gaatgtccag tgccatgcct gcattggagg caccaatgtt ggcgaggaca tcaggaagct
     1861 ggattacgga cagcatgttg tcgcgggcac tccagggcgt gtttttgata tgattcgtcg
     1921 cagaagccta aggacacgtg ctatcaaaat gttggttttg gatgaagctg atgaaatgtt
     1981 gaataaaggt ttcaaagagc agatttacga tgtatacagg tacctgcctc cagccacaca
     2041 ggtggttctc atcagtgcca cgctgccaca cgagattctg gagatgacca acaagttcat
     2101 gaccgaccca atccgcatct tggtgaaacg tgatgaattg actctggaag gcatcaagca
     2161 atttttcgtg gcagtggaga gggaagagtg gaaatttgac actctgtgtg acctctacga
     2221 cacactgacc atcactcagg cggtcatctt ctgcaacacc aaaagaaagg tggactggct
     2281 gacggagaaa atgagggaag ccaacttcac tgtatcctca atgcatggag acatgcccca
     2341 gaaagagcgg gagtccatca tgaaggagtt ccggtcgggc gccagccgag tgcttatttc
     2401 tacagatgtc tgggccaggg ggttggatgt ccctcaggtg tccctcatca ttaactatga
     2461 tctccctaat aacagagaat tgtacataca cagaattggg agatcaggtc gatacggccg
     2521 gaagggtgtg gccattaact ttgtaaagaa tgacgacatc cgcatcctca gagatatcga
     2581 gcagtactat tccactcaga ttgatgagat gccgatgaac gttgctgatc ttatctgaca
     2641 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     2701 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2761 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2821 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2881 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     2941 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3001 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3061 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3121 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3181 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3241 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3301 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3361 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3421 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3481 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3541 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3601 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3661 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3721 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3781 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3841 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3901 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3961 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4021 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4081 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4141 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4201 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4261 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4321 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4381 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4441 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4501 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4561 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4621 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4681 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4741 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4801 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4861 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4921 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4981 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5041 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5101 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5161 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5221 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5281 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5341 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5401 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5461 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5521 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5581 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5641 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5701 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5761 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5821 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5881 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5941 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6001 ccgccatgca t
//
