LOCUS       sequence-270809-        6377 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 6377)
  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 6377)
  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: pDB1 (CK2alpha)
FEATURES             Location/Qualifiers
     source          1..6377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..124
                     /gene="lacI"
                     /label=lacI promoter
     CDS             125..1207
                     /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     complement(144..163)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1256..1261,1262..1279,1280..1286)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1294..1310
                     /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)."
     primer_bind     1318..1334
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1318..1334
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1330..1857
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1350..1367)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1350..1366)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1359..1381)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1959..1964,1965..1980,1981..1987)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1995..2011
                     /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)."
     CDS             2034..2687
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2645..2667
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2697..2708
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4207..4229)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4370..4388)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4456..4560
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4561..4629,4630..5421)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4779..4798)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5592..6180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6081..6100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6334..6351
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca ccatcgaatg
       61 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
      121 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
      181 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
      241 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
      301 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
      361 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
      421 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
      481 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
      541 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
      601 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
      661 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
      721 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
      781 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
      841 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
      901 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
      961 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     1021 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     1081 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     1141 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     1201 gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccc caggctttac
     1261 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     1321 gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg tcgtgactgg
     1381 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1441 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1501 gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg gctggagtgc
     1561 gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat
     1621 gcgcccatct acaccaacgt gacctatccc attacggtca atccgccgtt tgttcccacg
     1681 gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg gctacaggaa
     1741 ggccagacgc gaattatttt tgatggcgtt ggaattagct tatcgactgc acggtgcacc
     1801 aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc
     1861 actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga
     1921 catcataacg gttctggcaa atattctgaa atgagctgtt gacaattaat catcggctcg
     1981 tataatgtgt ggaattgtga gcggataaca atttcacaca ggaaacagta ttcatgtccc
     2041 ctatactagg ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat
     2101 atcttgaaga aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa
     2161 acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg
     2221 ttaaattaac acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg
     2281 gtggttgtcc aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta
     2341 gatacggtgt ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc
     2401 ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt
     2461 taaatggtga tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt
     2521 tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta
     2581 ttgaagctat cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt
     2641 tgcagggctg gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatctgatcg
     2701 aaggtcgtgg gatcctgtcg ggacccgtgc caagcagggc cagagtttac acagatgtta
     2761 atacacacag acctcgagaa tactgggatt acgagtcaca tgtggtggaa tggggaaatc
     2821 aagatgacta ccagctggtt cgaaaattag gccgaggtaa atacagtgaa gtatttgaag
     2881 ccatcaacat cacaaataat gaaaaagttg ttgttaaaat tctcaagcca gtaaaaaaga
     2941 agaaaattaa gcgtgaaata aagattttgg agaatttgag aggaggtccc aacatcatca
     3001 cactggcaga cattgtaaaa gaccctgtgt cacgaacccc cgccttggtt tttgaacacg
     3061 taaacaacac agacttcaag caattgtacc agacgttaac agactatgat attcgatttt
     3121 acatgtatga gattctgaag gccctggatt attgtcacag catgggaatt atgcacagag
     3181 atgtcaagcc ccataatgtc atgattgatc atgagcacag aaagctacga ctaatagact
     3241 ggggtttggc tgagttttat catcctggcc aagaatataa tgtccgagtt gcttcccgat
     3301 acttcaaagg tcctgagcta cttgtagact atcagatgta cgattatagt ttggatatgt
     3361 ggagtttggg ttgtatgctg gcaagtatga tctttcggaa ggagccattt ttccatggac
     3421 atgacaatta tgatcagttg gtgaggatag ccaaggttct ggggacagaa gatttatatg
     3481 actatattga caaatacaac attgaattag atccacgttt caatgatatc ttgggcagac
     3541 actctcgaaa gcgatgggaa cgctttgtcc acagtgaaaa tcagcacctt gtcagccctg
     3601 aggccttgga tttcctggac aaactgctgc gatatgacca ccagtcacgg cttactgcaa
     3661 gagaggcaat ggagcacccc tatttctaca ctgttgtgaa ggaccaggct cgaatgggtt
     3721 catctagcat gccagggggc agtacgcccg tcagcagcgc caatatgatg tcagggattt
     3781 cttcagtgcc aaccccttca ccccttggac ctctggcagg ctcaccagtg attgctgctg
     3841 ccaaccccct tgggatgcct gttccagctg ccgctggcgc tcagcagtaa cggccctatc
     3901 tgtctcctga tgcctgagca gaggtggggg agtccaccct ctccttgatg cagcttgcgc
     3961 ctggcgggga ggggtgaaac acttcagaag caccgtgtct gaaccgttgc ttgtggattt
     4021 atagtagttc agtcataaaa aaaaaattat aataggctga ttttcttttt tctttttttt
     4081 tttaactcga acttttcata actcagggga ttccctgaaa aattacctgc agcccggggg
     4141 atccccggga attcatcgtg actgactgac gatctgcctc gcgcgtttcg gtgatgacgg
     4201 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4261 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     4321 catgacccag tcacgtagcg atagcggagt aattcttgaa gacgaaaggg cctcgtgata
     4381 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4441 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4501 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4561 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4621 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4681 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4741 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4801 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4861 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4921 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4981 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5041 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5101 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5161 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5221 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5281 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5341 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5401 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5461 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5521 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5581 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5641 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5701 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     5761 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5821 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5881 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5941 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6001 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6061 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6121 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6181 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6241 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6301 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6361 gagcgcctga tgcggta
//
