LOCUS       sequence-291133-        5602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HseIF2g_2-472.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5602)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5602)
  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-V5-SBP-C1-HseIF2g_2-472_AF
FEATURES             Location/Qualifiers
     source          1..5602
                     /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             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    2390..2511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2427..2446)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2481..2500
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2518..2973)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2655..2676)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2867..2886
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3000..3104
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3106..3463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3106..3297
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3129..3149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3314..3449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3498..4292
                     /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             3498..4292
                     /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(3552..3571)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4162..4181
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4480..4499)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4524..4571
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4900..5488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5389..5408
                     /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 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagcagcgg gcggagaagc tggagtgact ctagggcagc cgcatctttc
      901 gcgtcaggat ctcaccacct tggatgttac caagttgacg ccactttcac acgaagttat
      961 cagcagacaa gccacaatta acataggtac aattggtcat gtagctcatg ggaaatccac
     1021 agtcgtcaaa gctatttctg gagttcatac tgtcaggttc aaaaatgaac tagaaagaaa
     1081 tattacaatc aagcttggat atgctaatgc taagatttat aagcttgatg acccaagttg
     1141 ccctcggcca gaatgttata gatcttgtgg gagcagtaca cctgacgagt ttcctacgga
     1201 cattccaggg accaaaggga acttcaaatt agtcagacat gtttcctttg ttgactgtcc
     1261 tggccacgat attttgatgg ctactatgct gaacggtgca gcagtgatgg atgcagctct
     1321 tctgttgata gctggtaatg aatcttgccc tcagcctcag acatcggaac acctggctgc
     1381 tatagagatc atgaaactga agcatatttt gattctacaa aataaaattg atttggtaaa
     1441 agaaagtcag gctaaagaac aatacgagca gatccttgca tttgtccaag gtacagtagc
     1501 agagggagct cccattattc caatttcagc tcagctgaaa tacaatattg aagttgtttg
     1561 tgagtacata gtaaagaaaa ttccagtacc cccaagagac tttacttcag agccccggct
     1621 tattgttatt agatcttttg atgtcaacaa acctggctgt gaagttgatg accttaaggg
     1681 aggtgtagct ggtggtagta tcctaaaagg agtattaaag gtgggccagg agatagaagt
     1741 aagacctggt attgtttcca aagatagtga aggaaaactc atgtgtaaac caatcttttc
     1801 caaaattgta tcactttttg cggagcataa tgatctgcaa tatgctgctc caggcggtct
     1861 tattggagtt ggaacaaaaa ttgaccccac tttgtgccgg gctgacagaa tggtggggca
     1921 agtacttggt gcagtcggag ctttacctga gatattcaca gaattggaaa tttcctattt
     1981 cctgcttaga cggcttctag gtgtacgcac tgaaggagac aagaaagcag caaaggttca
     2041 aaagctgtct aagaatgaag tgctcatggt gaacatagga tccctgtcaa caggagggag
     2101 agttagtgct gtcaaggctg atttgggtaa aattgttttg accaatccag tgtgcacaga
     2161 ggtaggagaa aaaattgccc ttagccgaag agttgaaaaa cactggcgtt taattggttg
     2221 gggtcagata agaagaggag tgacaatcaa gccaacagta gatgatgact aatctagata
     2281 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2341 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2401 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2461 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     2521 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2581 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2641 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2701 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2761 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2821 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2881 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2941 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3001 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3061 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3121 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3181 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3241 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3301 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3361 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3421 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     3481 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3541 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3601 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3661 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3721 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3781 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3841 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3901 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3961 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4021 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4081 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4141 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4201 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4261 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4321 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4381 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4441 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     4501 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     4561 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4621 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4681 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     4741 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4801 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4861 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4921 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4981 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5041 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5101 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5161 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5221 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5281 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5341 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5401 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5461 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5521 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5581 taaccgtatt accgccatgc at
//
