LOCUS       sequence-290201-        7744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsXRN1_1-1173.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7744)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7744)
  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_1-1173_V
FEATURES             Location/Qualifiers
     source          1..7744
                     /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    4532..4653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4569..4588)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4623..4642
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4660..5115)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4797..4818)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5009..5028
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5142..5246
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5248..5605
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5248..5439
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5271..5291)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5456..5591
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5640..6434
                     /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             5640..6434
                     /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(5694..5713)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6304..6323
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6622..6641)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6666..6713
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7042..7630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7531..7550
                     /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 atgggagtcc ccaagtttta
      901 cagatggatc tcagagcggt atccctgtct cagcgaagtg gtgaaagagc atcagattcc
      961 tgaatttgac aacttgtacc tggatatgaa tggaattata catcagtgct cccatcctaa
     1021 tgatgatgat gttcacttta gaatttcaga tgataaaatc tttactgata tttttcacta
     1081 cctggaggtg ttgtttcgca ttattaaacc caggaaagtg ttctttatgg ctgtagatgg
     1141 tgtggctcct cgagcaaaaa tgaaccagca gcgtgggagg cgttttaggt cagcaaagga
     1201 ggcagaagac aaaattaaaa aggcaataga gaagggagaa actcttccta cagaggccag
     1261 atttgattcc aactgtatca caccaggaac tgaatttatg gccaggttac atgaacatct
     1321 gaagtatttt gtaaatatga aaatttccac agacaagtca tggcaaggag ttaccatcta
     1381 cttctcaggc catgagactc ctggagaagg agagcataaa atcatggaat ttatcagatc
     1441 cgagaaagca aagccagatc atgatccaaa caccagacac tgtctttatg gtttagatgc
     1501 tgacttgatt atgcttggat taacaagtca tgaggcacat ttttctctct taagagaaga
     1561 agttcgattt ggtggcaaaa aaacacaacg ggtatgtgct ccagaagaaa ctacatttca
     1621 ccttctacac ttgtctttaa tgagagagta tattgactat gagttttcag tattaaaaga
     1681 aaagatcaca tttaaatatg atattgaaag gataatagat gattggattt tgatggggtt
     1741 tcttgttggt aatgatttta tccctcatct acctcattta catattaatc atgatgcact
     1801 gcctcttctt tatggaacat atgttaccat cctgccagaa cttgggggtt atattaatga
     1861 aagtgggcac ctcaacttac ctcgatttga gaaatacctt gtgaaactat cagattttga
     1921 tcgggagcac ttcagtgaag tttttgtgga cctaaaatgg tttgaaagca aagttggtaa
     1981 caagtacctc aatgaagcag caggtgtcgc agcagaagaa gccaggaact acaaggaaaa
     2041 gaaaaagtta aagggccagg aaaattctct gtgttggact gctttagaca aaaatgaagg
     2101 cgaaatgata acttctaagg ataatttaga agatgagact gaagatgatg acctatttga
     2161 aactgagttt agacaatata aaagaacata ttacatgacg aagatggggg ttgacgtagt
     2221 atctgatgac tttctggctg atcaagctgc atgttatgtt caggcaatac agtggatttt
     2281 gcactattac tatcatggag ttcagtcctg gagctggtat tatccttatc attatgcgcc
     2341 tttcctgtct gatatacaca acatcagtac actcaaaatc cattttgaac taggaaaacc
     2401 ttttaagcca tttgaacagc ttcttgctgt acttccagca gccagcaaaa atttacttcc
     2461 tgcatgctac cagcatttga tgaccaatga agactcacca attatagaat attacccacc
     2521 tgattttaaa actgacctaa atgggaaaca acaggaatgg gaagctgtgg tgttaatccc
     2581 ttttattgat gagaagcgat tattggaagc catggagaca tgtaaccact ccctcaaaaa
     2641 ggaagagagg aaaagaaacc aacatagtga gtgcctaatg tgctggtatg atagagacac
     2701 agagtttatc tatccttctc catggccaga aaagttccct gccatagaac gatgttgtac
     2761 aaggtataaa ataatatcct tagatgcttg gcgtgtagac ataaacaaaa acaaaataac
     2821 cagaattgac cagaaagcat tatatttctg tggatttcct actctgaaac acatcagaca
     2881 caaatttttt ttgaagaaaa gtggtgttca agtattccag caaagcagtc gtggagaaaa
     2941 catgatgttg gaaatcttag tggatgcaga atcagatgaa cttaccgtag aaaatgtagc
     3001 ttcatcagtg cttggaaaat ctgtctttgt taattggcct caccttgagg aagctagagt
     3061 cgtggctgta tcagatggag aaactaagtt ttacttggaa gaacctccag gaacacagaa
     3121 gctttattca ggaagaactg ccccaccatc taaagtggtt catcttggag ataaagaaca
     3181 atctaactgg gcaaaagaag tacaaggaat ttcagaacac tacctgagaa gaaaaggaat
     3241 aataataaat gaaacatctg cagttgtgta tgctcagtta ctcacaggtc gtaaatatca
     3301 aataaatcaa aatggtgaag ttcgtctaga gaaacagtgg tcaaaacaag ttgttccttt
     3361 tgtttatcaa actattgtca aggacatccg agctttcgac tcccgtttct ccaatatcaa
     3421 aacattggat gatttgtttc ctctgagaag tatggtcttt atgctgggaa ctccctatta
     3481 tggctgcact ggagaagttc aggattcagg tgatgtgatt acagaaggta ggattcgtgt
     3541 gattttcagc attccatgtg aacccaatct tgatgcttta atacagaacc agcataaata
     3601 ttctataaag tacaacccag gatatgtgtt ggccagtcgc cttggagtga gtggatacct
     3661 tgtttcaagg tttacaggaa gtatttttat tggaagagga tctaggagaa accctcatgg
     3721 agaccataaa gcaaatgtgg gtttaaatct caaattcaac aagaaaaatg aggaggtacc
     3781 tggatatact aagaaagttg gaagtgaatg gatgtattca tctgcagcag aacaacttct
     3841 ggcagagtac ttagagagag ctccagaact atttagttat atagccaaaa atagccaaga
     3901 ggatgtgttc tatgaagatg acatttggcc tggagaaaat gagaatggtg ctgaaaaagt
     3961 tcaagaaatt attacttggc taaaaggaca tcctgtcagt actttatctc gttcttcttg
     4021 tgatttacaa attctggatg cagctattgt tgagaaaatt gaggaagaag tcgaaaagtg
     4081 caagcaaaga aagaataata agaaggtgcg agtaacagtg aaaccccatt tgctatacag
     4141 acctttagaa cagcaacatg gagtcattcc tgatcgggat gcagaatttt gtctttttga
     4201 ccgtgttgta aatgtgagag aaaacttctc agttccagtt ggccttcgag gcaccatcat
     4261 aggaataaaa ggagctaata gagaagccga tgtactattt gaagtattat ttgatgaaga
     4321 atttcctgga gggttaacaa taagatgctc acctggtaga ggttatcgac tgccaacaag
     4381 tgccttggtg aacctttctt aaggatccac cggatctaga taactgatca taatcagcca
     4441 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4501 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4561 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4621 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     4681 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4741 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4801 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4861 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4921 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4981 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5041 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     5101 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5161 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5221 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     5281 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5341 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5401 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5461 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5521 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5581 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     5641 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5701 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5761 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5821 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5881 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5941 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6001 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6061 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6121 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     6181 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6241 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6301 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6361 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6421 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     6481 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6541 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6601 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     6661 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     6721 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6781 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     6841 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     6901 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6961 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7021 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     7201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7681 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     7741 gcat
//
