LOCUS       sequence-290944-        5701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsMex3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5701)
  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-HsMex3C_AD
FEATURES             Location/Qualifiers
     source          1..5701
                     /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     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"
     misc_feature    845..901
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2489..2610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2526..2545)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2580..2599
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2617..3072)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2754..2775)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2966..2985
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3099..3203
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3205..3562
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3205..3396
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3228..3248)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3413..3548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3597..4391
                     /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             3597..4391
                     /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(3651..3670)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4261..4280
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4579..4598)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4623..4670
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4999..5587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5488..5507
                     /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 ggtaccgcgg gcccgggatc
      901 catgcccagc ggcagctccg cggccctggc cccggcggcg gcgggggtgc tgtacggagg
      961 ggacgatgcc cagggcatga tggcggcgat gctgtcccac gcctacggcc ccggcggttg
     1021 tggggcggcg gcggccgccc tgaacgggga gcaggcggcc ctgctccgga gaaagagcgt
     1081 caacaccacc gagtgcgtcc cggtgcccag ctccgagcac gtcgccgaga tcgtcggccg
     1141 ccagggttgt aaaattaaag cactgagagc caagacaaac acgtatatca agactcctgt
     1201 tcgtggtgaa gagcccattt ttgttgtcac tggaaggaaa gaagatgttg ccatggccaa
     1261 aagagagatc ctctcagctg cagagcactt ctccatgatt cgtgcatctc gaaacaaaaa
     1321 tgggcctgcc ctgggaggat tatcatgtag tcctaatctg cccggtcaaa ccaccgtcca
     1381 agtcagggtc ccttatcgtg tggtaggatt agtggttgga cccaaaggag caactattaa
     1441 aagaattcag cagcagaccc acacctacat agtaactccg agcagagata aggaacctgt
     1501 ctttgaagtg acagggatgc ctgaaaatgt tgaccgagca cgggaagaaa tagaaatgca
     1561 tattgccatg cgtacaggaa actatataga gctcaatgaa gagaatgatt tccattacaa
     1621 tggtaccgat gtaagctttg aaggtggcac tcttggctct gcgtggctct cctccaatcc
     1681 tgttcctcct agccgcgcaa gaatgatatc caattatcga aatgatagtt ccagttctct
     1741 aggaagtggc tctacagatt cctactttgg aagcaatagg ctggctgact ttagtccaac
     1801 aagcccattt agcacaggaa acttctggtt tggagataca ctaccatctg taggctcaga
     1861 agacctagca gttgactctc ctgcctttga ctctttacca acatctgctc aaactatctg
     1921 gactccattt gaaccagtta acccactctc tggctttggg agtgatcctt ctggtaacat
     1981 gaagactcag cgcagaggaa gtcagccatc tactcctcgt ctgtctccta catttcctga
     2041 gagcatagaa catccacttg ctcggagggt taggagcgac ccacctagta caggcaacca
     2101 tgttggcctt ccaatatata tccctgcttt ttctaatggt accaatagtt actcctcttc
     2161 caatggtggt tccacctcta gctcacctcc agaatcaaga cgaaagcacg actgtgtgat
     2221 ttgctttgag aatgaggtta ttgctgccct agttccatgt ggccacaacc tcttctgcat
     2281 ggaatgtgcc aacaagatct gtgaaaagag aacgccatca tgtccagttt gccagacagc
     2341 tgttactcag gcaatccaaa ttcactctta atctagataa ctgatcataa tcagccatac
     2401 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2461 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2521 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2581 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     2641 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2701 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2761 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2821 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2881 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2941 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3001 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3061 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3121 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3181 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3241 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3301 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3361 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3421 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3481 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3541 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3601 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3661 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3721 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     3781 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3841 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3901 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3961 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4021 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4081 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4141 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4201 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4261 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4321 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4381 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4441 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4501 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4561 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     4621 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     4681 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4741 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4801 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     4861 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4921 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4981 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5041 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5101 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     5161 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5221 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5281 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5341 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5401 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5461 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5521 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5581 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5641 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     5701 t
//
