LOCUS       sequence-486747-        8997 bp    DNA     circular SYN 19-SEP-2025
DEFINITION  Mammalian Expression of HseIF4G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8997)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8997)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLNHA-C1-HseIF4G_O
FEATURES             Location/Qualifiers
     source          1..8997
                     /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             665..730
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             755..781
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5785..5906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5822..5841)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5876..5895
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5913..6368)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6050..6071)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6262..6281
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6395..6499
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6501..6858
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6501..6692
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6524..6544)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6709..6844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6893..7687
                     /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             6893..7687
                     /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(6947..6966)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7557..7576
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7875..7894)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7919..7966
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8295..8883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8784..8803
                     /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 caccatggac gcacaaacac gacgacgtga gcgtcgcgct gagaaacaag ctcaatggaa
      721 agctgcaaac ggtggatctc ctccacatat ggcttaccca tacgatgttc cagattacgc
      781 tcctccatct cgagctatga acaaagctcc acagtccaca ggccccccac ccgccccatc
      841 ccccggactc ccacagccag cgtttccccc ggggcagaca gcgccggtgg tgttcagtac
      901 gccacaagcg acacaaatga acacgccttc tcagccccgc cagggaggat tcaggtctct
      961 gcagcacttc taccctagcc gggcccagcc cccgagcagt gcagcctccc gagtgcagag
     1021 tgcagcccct gcccgccctg gcccagctgc ccatgtctac cctgctggat cccaagtaat
     1081 gatgatccct tcccagatct cctacccagc ctcccagggg gcctactaca tccctggaca
     1141 ggggcgttcc acatacgttg tcccgacaca gcagtaccct gtgcagccag gagccccagg
     1201 cttctatcca ggtgcaagcc ctacagaatt tgggacctac gctggcgcct actatccagc
     1261 ccaaggggtg cagcagtttc ccactggcgt ggcccccgcc ccagttttga tgaaccagcc
     1321 accccagatt gctcccaaga gggagcgtaa gacgatccga attcgagatc caaaccaagg
     1381 aggaaaggat atcacagagg agatcatgtc tggggcccgc actgcctcca cacccacccc
     1441 tccccagacg ggaggcggtc tggagcctca agctaatggg gagacgcccc aggttgctgt
     1501 cattgtccgg ccagatgacc ggtcacaggg agcaatcatt gctgaccggc cagggctgcc
     1561 tggcccagag catagccctt cagaatccca gccttcgtcg ccttctccga ccccatcacc
     1621 atccccagtc ttggaaccgg ggtctgagcc taatctcgca gtcctctcta ttcctgggga
     1681 cactatgaca actatacaaa tgtctgtaga agaatcaacc cccatctccc gtgaaactgg
     1741 ggagccatat cgcctctctc cagaacccac tcctctcgcc gaacccatac tggaagtaga
     1801 agtgacactt agcaaaccgg ttccagaatc tgagttttct tccagtcctc tccaggctcc
     1861 cacccctttg gcatctcaca cagtggaaat tcatgagcct aatggcatgg tcccatctga
     1921 agatctggaa ccagaggtgg agtcaagccc agagcttgct cctcccccag cttgcccctc
     1981 cgaatcccct gtgcccattg ctccaactgc ccaacctgag gaactgctca acggagcccc
     2041 ctcgccacca gctgtggact taagcccagt cagtgagcca gaggagcagg ccaaggaggt
     2101 gacagcatca gtggcgcccc ccaccatccc ctctgctact ccagctacgg ctccttcagc
     2161 tacttcccca gctcaggagg aggaaatgga agaagaagaa gaagaggaag aaggagaagc
     2221 aggagaagca ggagaagctg agagtgagaa aggaggagag gaactgctcc ccccagagag
     2281 tacccctatt ccagccaact tgtctcagaa tttggaggca gcagcagcca ctcaagtggc
     2341 agtatctgtg ccaaagagga gacggaaaat taaggagcta aataagaagg aggctgttgg
     2401 agaccttctg gatgccttca aggaggcgaa cccggcagta ccagaggtgg aaaatcagcc
     2461 tcctgcaggc agcaatccag gcccagagtc tgagggcagt ggtgtgcccc cacgtcctga
     2521 ggaagcagat gagacctggg actcaaagga agacaaaatt cacaatgctg agaacatcca
     2581 gcccggggaa cagaagtatg aatataagtc agatcagtgg aagcctctaa acctagagga
     2641 gaaaaaacgt tacgaccgtg agttcctgct tggttttcag ttcatctttg ccagtatgca
     2701 gaagccagag ggattgccac atatcagtga cgtggtgctg gacaaggcca ataaaacacc
     2761 actgcggcca ctggatccca ctagactaca aggcataaat tgtggcccag acttcactcc
     2821 atcctttgcc aaccttggcc ggacaaccct tagcacccgt gggcccccaa ggggtgggcc
     2881 aggtggggag ctgccccgtg ggccggctgg cctgggaccc cggcgctctc agcagggacc
     2941 ccgaaaagaa ccacgcaaga tcattgccac agtgttaatg accgaagata taaaactgaa
     3001 caaagcagag aaagcctgga aacccagcag caagcggacg gcggctgata aggatcgagg
     3061 ggaagaagat gctgatggca gcaaaaccca ggacctattc cgcagggtgc gctccatcct
     3121 gaataaactg acaccccaga tgttccagca gctgatgaag caagtgacgc agctggccat
     3181 cgacaccgag gaacgcctca aaggggtcat tgacctcatt tttgagaagg ccatttcaga
     3241 gcccaacttc tctgtggcct atgccaacat gtgccgctgc ctcatggcgc tgaaagtgcc
     3301 cactacggaa aagccaacag tgactgtgaa cttccgaaag ctgttgttga atcgatgtca
     3361 gaaggagttt gagaaagaca aagatgatga tgaggttttt gagaagaagc aaaaagagat
     3421 ggatgaagct gctacggcag aggaacgagg acgcctgaag gaagagctgg aagaggctcg
     3481 ggacatagcc cggcggcgct ctttagggaa tatcaagttt attggagagt tgttcaaact
     3541 gaagatgtta acagaggcaa taatgcatga ctgtgtggtc aaactgctta agaaccatga
     3601 tgaagagtcc cttgagtgcc tttgtcgtct gctcaccacc attggcaaag acctggactt
     3661 tgaaaaagcc aagccccgaa tggatcagta tttcaaccag atggaaaaaa tcattaaaga
     3721 aaagaagacg tcatcccgca tccgctttat gctgcaggac gtgctggatc tgcgagggag
     3781 caattgggtg ccacgccgag gggatcaggg tcccaagacc attgaccaga tccataagga
     3841 ggctgagatg gaagaacatc gagagcacat caaagtgcag cagctcatgg ccaagggcag
     3901 tgacaagcgt cggggcggtc ctccaggccc tcccatcagc cgtggacttc cccttgtgga
     3961 tgatggtggc tggaacacag ttcccatcag caaaggtagc cgccccattg acacctcacg
     4021 actcaccaag atcaccaagc ctggctccat cgattctaac aaccagctct ttgcacctgg
     4081 agggcgactg agctggggca agggcagcag cggaggctca ggagccaagc cctcagacgc
     4141 agcatcagaa gctgctcgcc cagctactag tactttgaat cgcttctcag cccttcaaca
     4201 agcggtaccc acagaaagca cagataatag acgtgtggtg cagaggagta gcttgagccg
     4261 agaacgaggc gagaaagctg gagaccgagg agaccgccta gagcggagtg aacggggagg
     4321 ggaccgtggg gaccggcttg atcgtgcgcg gacacctgct accaagcgga gcttcagcaa
     4381 ggaagtggag gagcggagta gagaacggcc ctcccagcct gaggggctgc gcaaggcagc
     4441 tagcctcacg gaggatcggg accgtgggcg ggatgccgtg aagcgagaag ctgccctacc
     4501 cccagtgagc cccctgaagg cggctctctc tgaggaggag ttagagaaga aatccaaggc
     4561 tatcattgag gaatatctcc atctcaatga catgaaagag gcagtccagt gcgtgcagga
     4621 gctggcctca ccctccttgc tcttcatctt tgtacggcat ggtgtcgagt ctacgctgga
     4681 gcgcagtgcc attgctcgtg agcatatggg gcagctgctg caccagctgc tctgtgctgg
     4741 gcatctgtct actgctcagt actaccaagg gttgtatgaa atcttggaat tggctgagga
     4801 catggaaatt gacatccccc acgtgtggct ctacctagcg gaactggtaa cacccattct
     4861 gcaggaaggt ggggtgccca tgggggagct gttcagggag attacaaagc ctctgagacc
     4921 gttgggcaaa gctgcttccc tgttgctgga gatcctgggc ctcctgtgca aaagcatggg
     4981 tcctaaaaag gtggggacgc tgtggcgaga agccgggctt agctggaagg aatttctacc
     5041 tgaaggccag gacattggtg cattcgtcgc tgaacagaag gtggagtata ccctgggaga
     5101 ggagtcggaa gcccctggcc agagggcact cccctccgag gagctgaaca ggcagctgga
     5161 gaagctgctg aaggagggca gcagtaacca gcgggtgttc gactggatag aggccaacct
     5221 gagtgagcag cagatagtat ccaacacgtt agttcgagcc ctcatgacgg ctgtctgcta
     5281 ttctgcaatt atttttgaga ctcccctccg agtggacgtt gcagtgctga aagcgcgagc
     5341 gaagctgctg cagaaatacc tgtgtgacga gcagaaggag ctacaggcgc tctacgccct
     5401 ccaggccctt gtagtgacct tagaacagcc tcccaacctg ctgcggatgt tctttgacgc
     5461 actgtatgac gaggacgtgg tgaaggagga tgccttctac agttgggaga gtagcaagga
     5521 ccccgctgag cagcagggca agggtgtggc ccttaaatct gtcacagcct tcttcaagtg
     5581 gctccgtgaa gcagaggagg agtctgacca caactgaaag cttcgaattc tgcagtcgac
     5641 ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag ccataccaca
     5701 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5761 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5821 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5881 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     5941 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6001 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6061 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6121 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6181 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6241 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6301 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6361 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6421 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6481 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     6541 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6601 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6661 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6721 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6781 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6841 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     6901 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     6961 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     7021 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     7081 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     7141 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     7201 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     7261 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     7321 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     7381 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     7441 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     7501 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     7561 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     7621 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     7681 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     7741 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     7801 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     7861 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     7921 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     7981 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     8041 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     8101 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     8161 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8221 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8281 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8341 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8401 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8461 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8521 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8581 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8641 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8701 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8761 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8821 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8881 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8941 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
