LOCUS       sequence-291131-        5143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HseIF2a_2-315.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5143)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5143)
  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-HseIF2a_2-315_AF
FEATURES             Location/Qualifiers
     source          1..5143
                     /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    1931..2052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1968..1987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2022..2041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2059..2514)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2196..2217)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2408..2427
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2541..2645
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2647..3004
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2647..2838
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2670..2690)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2855..2990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3039..3833
                     /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             3039..3833
                     /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(3093..3112)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3703..3722
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4021..4040)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4065..4112
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4441..5029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4930..4949
                     /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 cgagcaccgg gtctaagttg tagattttat caacacaaat ttcctgaggt
      901 ggaagatgta gtgatggtga atgtcagatc cattgctgaa atgggggctt atgtcagctt
      961 gctggaatac aacaacattg aaggcatgat tcttcttagt gaattatcca gaaggcgtat
     1021 ccgttctatc aacaaactca tccgaattgg caggaatgag tgtgtggttg tcattagggt
     1081 ggacaaagaa aaaggatata ttgatttgtc aaaaagaaga gtttctccag aggaagcaat
     1141 caaatgtgaa gacaaattca caaaatccaa aactgtttat agcattcttc gtcatgttgc
     1201 tgaggtgtta gaatacacca aggatgagca gctggaaagc ctattccaga ggactgcctg
     1261 ggtctttgat gacaagtaca agagacctgg atatggtgcc tatgatgcat ttaagcatgc
     1321 agtctcagac ccatctattt tggatagttt agatttgaat gaagatgaac gggaagtact
     1381 cattaataat attaataggc gcttgacccc acaggctgtc aaaattcgag cagatattga
     1441 agtggcttgt tatggttatg aaggcattga tgctgtaaaa gaagccctaa gagcaggttt
     1501 gaattgttct acagaaaaca tgcccattaa gattaatcta atagctcctc ctcggtatgt
     1561 aatgactacg acaaccctgg agagaacaga aggcctttct gtcctcagtc aagctatggc
     1621 tgttatcaaa gagaagattg aggaaaagag gggtgtgttc aatgttcaaa tggagcccaa
     1681 agtggtcaca gatacagatg agactgaact tgcgaggcag atggagaggc ttgaaagaga
     1741 aaatgccgaa gtggatggag atgatgatgc agaagaaatg gaagccaaag ctgaagatta
     1801 aggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     1861 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     1921 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1981 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2041 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2101 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2161 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2221 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2281 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2341 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2401 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2461 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2521 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2581 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2641 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2701 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2761 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2821 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2881 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2941 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3001 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3061 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3121 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3181 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3241 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3301 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3361 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3421 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3481 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3541 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     3601 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3661 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3721 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3781 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3841 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3901 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3961 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4021 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4081 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4141 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4201 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4261 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     4321 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4381 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4441 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4501 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4561 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4621 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4681 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4741 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4801 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4861 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4921 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4981 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5041 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5101 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
