LOCUS       sequence-357454-        5827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsXRN1_1174-1706.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5827)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5827)
  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-HsXRN1_1174-1706_V
FEATURES             Location/Qualifiers
     source          1..5827
                     /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    2615..2736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2652..2671)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2706..2725
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2743..3198)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2880..2901)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3092..3111
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3225..3329
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3331..3688
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3331..3522
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3354..3374)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3539..3674
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3723..4517
                     /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             3723..4517
                     /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(3777..3796)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4387..4406
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4705..4724)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4749..4796
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5125..5713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5614..5633
                     /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 cgagctcaag cttcgaattc tgcagtcgac atgcatggga gtcgctctga
      901 aactggaaat cagaagttga cagccatcgt aaaaccacaa ccagctgtac atcaacatag
      961 ctcaagttca tcagtttcct ctgggcattt gggagccctc aaccattccc ctcaatcact
     1021 ttttgttcct actcaagtac ctactaaaga tgatgatgaa ttctgcaaca tttggcagtc
     1081 cttacaggga tctggaaaga tgcaatactt tcagccaact atacaagaga agggtgcagt
     1141 tctacctcaa gaaataagcc aagtaaatca acatcataaa tctggcttta atgacaacag
     1201 tgttaaatat cagcaaagaa aacatgaccc tcacagaaaa tttaaagaag agtgtaagag
     1261 tcctaaagct gagtgttggt cccaaaaaat gtccaataag cagcctaact ctggaattga
     1321 gaacttttta gcatctttga atatctccaa agaaaatgaa gtacagtcat ctcatcatgg
     1381 ggagcctcca agtgaagagc atttgtcacc acagtcattt gccatgggaa cacggatgct
     1441 taaagaaatt ctaaaaattg atggctctaa cactgtggac cataagaatg aaatcaaaca
     1501 gattgctaat gaaatccctg tttcctctaa cagaagagat gaatatggat taccctctca
     1561 gcctaaacaa aataagaaat tagcatctta tatgaacaag cctcacagtg ctaatgagta
     1621 ccataatgtt cagtctatgg acaatatgtg ttggcctgcc cccagccaga tccctcctgt
     1681 atccacacca gtaactgaac tttctcgaat ttgttccctt gttggaatgc cacaacctga
     1741 tttctccttt cttaggatgc cacagacaat gaccgtttgc caagtaaaat tatctaatgg
     1801 cttactggta catgggccac agtgccactc tgaaaatgaa gccaaagaga aagctgcact
     1861 ttttgcttta caacagttgg gctccttagg catgaatttc cctttgcctt cacaagtatt
     1921 tgcaaattat ccttcagctg taccacctgg aaccattcct ccagcctttc ccccacctac
     1981 tggctgggat cactatggaa gcaactatgc attgggggca gctaatataa tgccttcgtc
     2041 gtctcatctc tttggctcaa tgccatgggg accatcggtg ccagttcctg ggaagccctt
     2101 ccatcatact ttatattctg ggaccatgcc catggctggg ggaataccag ggggtgtgca
     2161 caatcagttt atacctctgc aggttactaa aaaaagggtt gcaaacaaaa agaactttga
     2221 gaataaggaa gcccagagtt ctcaagccac tccagttcag actagccagc cagattcttc
     2281 caacattgtc aaagtaagtc cacgggagag ctcatcagct tctttgaagt cctctccgat
     2341 tgctcaacct gcatcttctt ttcaagttga aactgcctct caaggccata gtatatctca
     2401 ccataagtca acaccaatct cttcttcaag aagaaaatca agaaaactgg ctgttaattt
     2461 tggtgtttct aaaccttctg agtaaggatc caccggatct agataactga tcataatcag
     2521 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2581 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2641 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2701 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     2761 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2821 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2881 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2941 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3001 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3061 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3121 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3181 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3241 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3301 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3361 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3421 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3481 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3541 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3601 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3661 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3721 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3781 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3841 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3901 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3961 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4021 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4081 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4141 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4201 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4261 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4321 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4381 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4441 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4501 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4561 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4621 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4681 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4741 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4801 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4861 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4921 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     4981 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5041 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5101 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5161 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5221 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5281 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5341 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5401 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5461 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5521 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5581 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5641 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5701 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5761 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5821 catgcat
//
