LOCUS       sequence-289173-        5981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsRck-mut1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5981)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5981)
  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: pCMV-TAPtagN-HsRCK-mut1_K
FEATURES             Location/Qualifiers
     source          1..5981
                     /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      658..667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..867
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             868..1041
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1078..1098
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1105..1182
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    2769..2890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2806..2825)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2860..2879
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2897..3352)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3034..3055)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3246..3265
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3379..3483
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3485..3842
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3485..3676
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3508..3528)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3693..3828
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3877..4671
                     /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             3877..4671
                     /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(3931..3950)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4541..4560
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4859..4878)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4903..4950
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5279..5867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5768..5787
                     /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 taccggtgcc
      661 accatggcag gccttgcgca acacgatgaa gccgtggaca acaaattcaa caaagaacaa
      721 caaaacgcgt tctatgagat cttacattta cctaacttaa acgaagaaca acgaaacgcc
      781 ttcatccaaa gtttaaaaga tgacccaagc caaagcgcta accttttagc agaagctaaa
      841 aagctaaatg atgctcaggc gccgaaagta gacaacaaat tcaacaaaga acaacaaaac
      901 gcgttctatg agatcttaca tttacctaac ttaaacgaag aacaacgaaa cgccttcatc
      961 caaagtttaa aagatgaccc aagccaaagc gctaaccttt tagcagaagc taaaaagcta
     1021 aatgatgctc aggcgccgaa agtagacgcg aattgtgata tacctacaac tgcttctgaa
     1081 aatttatatt ttcaaggtga actaaagaga agatggaaaa agaatttcat agccgtctca
     1141 gcagccaacc gctttaagaa aatctcatcc tccggggcac ttgatccatc cggactcaga
     1201 tctcgagcta tgggtctgtc cagtcaaaat ggtcagctga gaggccctgt gaaacccact
     1261 ggtggccctg gaggaggggg cacacagaca cagcaacaga tgaaccagct gaaaaacacc
     1321 aacacaatca ataatggcac tcagcagcaa gcacagagta tgaccaccac tattaaacct
     1381 ggtgatgact ggaaaaagac tttaaaactc cctccaaagg atctaagaat caaaacttcg
     1441 gatgtgacct ccacaaaagg aaatgagttt gaagattact gtttgaaacg ggagttactg
     1501 atgggaattt ttgaaatggg ctgggaaaag ccatctccta ttcaggagga gagcattccc
     1561 attgctttat ctggtaggga tatcttagct agagcaaaaa atggaacagg caagagcggt
     1621 gcctacctca ttcccttact tgaacggcta gacctgaaga aggacaatat acaagcaatg
     1681 gtgattgttc ccactagaga acttgctcta caggtcagtc aaatttgcat ccaggtcagc
     1741 aaacacatgg gaggggccaa agtgatggca accacaggag gaaccaattt acgagatgac
     1801 ataatgaggc ttgatgatac agtgcacgtg gtgattgcta cccctgggag aatcctggat
     1861 cttattaaga aaggagtagc aaaggttgat catgtccaga tgatagtatt ggatgaggca
     1921 gataagttgc tgtcacagga ttttgtgcag ataatggagg atattattct cacgctacct
     1981 aaaaacaggc agattttact atattccgct actttccctc ttagtgtaca gaagttcatg
     2041 aattcccatt tgcagaaacc ctatgagatt aacctgatgg aggaactaac tctgaaggga
     2101 gtaacccagt actacgcata tgtaactgag cgcgcaaaag tagcatgcct caacgcactt
     2161 ttctccgcac ttcagataaa ccagtcgatc attttctgta actcctctca gcgagttgaa
     2221 ttgctagcca agaagatttc tcaactgggt tattcttgct tctatattca tgctaaaatg
     2281 aggcaggaac atcgaaatcg tgtatttcat gatttccgaa atggcttatg ccgcaatctt
     2341 gtttgcactg atctgtttac ccgaggtatt gatatacaag ctgtgaatgt ggtaataaac
     2401 tttgatttcc caaagctggc agagacctat ctccatcgta ttggaagatc aggtcgcttt
     2461 ggtcatcttg gcttagccat caacttgatc acatatgatg atcgcttcaa cctgaaaagt
     2521 attgaggagc agctgggaac agaaattaaa cctattccga gcaacattga taagagcctg
     2581 tatgtggcag aataccacag cgagcctgta gaagatgaga aaccttaacc gcgggcccgg
     2641 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     2701 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2761 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2821 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2881 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2941 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3001 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3061 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3121 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3181 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3241 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3301 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3361 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3421 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3481 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3541 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3601 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3661 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3721 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3781 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3841 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3901 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3961 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4021 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4081 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4141 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4201 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4261 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4321 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4381 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4441 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4501 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4561 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4621 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4681 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4741 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4801 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4861 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4921 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4981 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5041 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5101 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5161 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5221 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5281 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5341 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5401 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5461 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5521 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5581 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5641 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5701 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5761 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5821 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5881 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5941 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
