LOCUS       sequence-133114-        6148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6148)
  AUTHORS   Turowec JP, Duncan JS, French AC, Gyenis L, St Denis NA, Vilk G, 
            Litchfield DW.
  TITLE     Protein kinase CK2 is a constitutively active enzyme that promotes 
            cell survival: strategies to identify CK2 substrates and manipulate 
            its activity in mammalian cells.
  JOURNAL   Methods Enzymol. 2010;484:471-93.
  PUBMED    21036246
REFERENCE   2  (bases 1 to 6148)
  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"; journalName: "Methods 
            Enzymol."; date: "2010"; volume: "484"; pages: "471-93"
COMMENT     Sequence Label: pAB46 (CK2beta)
FEATURES             Location/Qualifiers
     source          1..6148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..37)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     178..200
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(194..385)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1194..2276)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2238..2257
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2277..2354)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2560..2579
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2663..2682
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2663..2681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2682..2706
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             2721..2743
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2762..2779
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2789..2806
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2810..2842
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             3675..3692
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3745..3763)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3759..3806
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3843..4298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3930..3949)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4140..4161
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4390..5205)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4390..5205)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5113..5132
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5197..5216
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5327..5915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5816..5835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6069..6086
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(6101..6148,1..92)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
       61 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
      121 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      181 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
      241 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
      301 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
      361 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
      421 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
      481 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
      541 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
      601 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
      661 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
      721 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
      781 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
      841 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
      901 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
      961 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     1021 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     1081 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     1141 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     1201 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1261 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     1321 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     1381 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     1441 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     1501 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     1561 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     1621 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     1681 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     1741 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     1801 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     1861 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     1921 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     1981 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     2041 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     2101 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     2161 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     2221 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     2281 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     2341 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     2401 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     2461 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     2521 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     2581 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     2641 cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
     2701 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca tgggcagcag
     2761 ccatcatcat catcatcaca gcagcggcct ggtgccgcgc ggcagccata tggctagcat
     2821 gactggtgga cagcaaatgg gtcgcggatc cgaattcgag ctccgtcgac aagcttgcgg
     2881 ccgcagcagc tcagaggagg tgtcctggat ttcctggttc tgtgggctcc gtggcaatga
     2941 attcttctgt gaagtggatg aagactacat ccaggacaaa tttaatctta ctggactcaa
     3001 tgagcaggtc cctcactatc gacaagctct agacatgatc ttggacctgg agcctgatga
     3061 agaactggaa gacaacccca accagagtga cctgattgag caggcagccg agatgcttta
     3121 tggattgatc cacgcccgct acatccttac caaccgtggc atcgcccaga tgttggaaaa
     3181 gtaccagcaa ggagactttg gttactgtcc tcgtgtgtac tgtgagaacc agccaatgct
     3241 tcccattggc ctttcagaca tcccaggtga agccatggtg aagctctact gccccaagtg
     3301 catggatgtg tacacaccca agtcatcaag acaccatcac acggatggcg cctacttcgg
     3361 cactggtttc cctcacatgc tcttcatggt gcatcccgag taccggccca agagacctgc
     3421 caaccagttt gtgcccaggc tctacggttt caagatccat ccgatggcct accagctgca
     3481 gctccaagcc gccagcaact tcaagagccc agtcaagacg attcgctgat tccctccccc
     3541 acctgtcctg cagtctggat cccccgggct gcaggaattc gatatcaagc ttggtaccga
     3601 gctcggatcc actagtaacg gccgccagtg tgctggaatt ctgcagatat ccatcacact
     3661 ggcggccgct cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     3721 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     3781 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     3841 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3901 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3961 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4021 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4081 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4141 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4201 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4261 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4321 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4381 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     4441 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     4501 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     4561 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     4621 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     4681 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     4741 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     4801 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     4861 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     4921 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     4981 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     5041 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     5101 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     5161 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     5221 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     5281 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5341 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5401 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5461 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5521 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5581 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5641 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5701 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5761 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5821 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5881 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5941 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6001 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6061 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     6121 ttacgcatct gtgcggtatt tcacaccg
//
