LOCUS       Exported                6909 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pZW6 (CK2alpha)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6909)
  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 6909)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6909
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     691..707
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     691..707
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1976..2065
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     promoter        complement(2134..2152)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2135..2152)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2172..2189)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2178..2402
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2448..2961
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2535..2554)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2745..2766
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2984..3004)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2989..3318
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3169..3304
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3231..3250
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3385..4179
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3439..3458)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4049..4068
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4353..4487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4390..4409)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4444..4463
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4518..4534)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4518..4534)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4531..4553)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4542..4558
                     /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)."
     promoter        complement(4566..4596)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4611..4632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4749..4766)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4920..5508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5000..5019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5679..6539)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6302..6321
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6540..6644)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6719..6738)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tccaccgcgg tggcggccgc tctagaacta gtggatcctc tgaaacaagg
      721 tcttaccccc agctgcttct gaacacagtg actgccagat ctccaaacat caagtccagc
      781 tttgtccgcc aacctgtctg acatgtcggg acccgtgcca agcagggcca gagtttacac
      841 agatgttaat acacacagac ctcgagaata ctgggattac gagtcacatg tggtggaatg
      901 gggaaatcaa gatgactacc agctggttcg aaaattaggc cgaggtaaat acagtgaagt
      961 atttgaagcc atcaacatca caaataatga aaaagttgtt gttaaaattc tcaagccagt
     1021 aaaaaagaag aaaattaagc gtgaaataaa gattttggag aatttgagag gaggtcccaa
     1081 catcatcaca ctggcagaca ttgtaaaaga ccctgtgtca cgaacccccg ccttggtttt
     1141 tgaacacgta aacaacacag acttcaagca attgtaccag acgttaacag actatgatat
     1201 tcgattttac atgtatgaga ttctgaaggc cctggattat tgtcacagca tgggaattat
     1261 gcacagagat gtcaagcccc ataatgtcat gattgatcat gagcacagaa agctacgact
     1321 aatagactgg ggtttggctg agttttatca tcctggccaa gaatataatg tccgagttgc
     1381 ttcccgatac ttcaaaggtc ctgagctact tgtagactat cagatgtacg attatagttt
     1441 ggatatgtgg agtttgggtt gtatgctggc aagtatgatc tttcggaagg agccattttt
     1501 ccatggacat gacaattatg atcagttggt gaggatagcc aaggttctgg ggacagaaga
     1561 tttatatgac tatattgaca aatacaacat tgaattagat ccacgtttca atgatatctt
     1621 gggcagacac tctcgaaagc gatgggaacg ctttgtccac agtgaaaatc agcaccttgt
     1681 cagccctgag gccttggatt tcctggacaa actgctgcga tatgaccacc agtcacggct
     1741 tactgcaaga gaggcaatgg agcaccccta tttctacact gttgtgaagg accaggctcg
     1801 aatgggttca tctagcatgc cagggggcag tacgcccgtc agcagcgcca atatgatgtc
     1861 agggatttct tcagtgccaa ccccttcacc ccttggacct ctggcaggct caccagtgat
     1921 tgctgctgcc aacccccttg ggatgcctgt tccagctgcc gctggcgctc agcagtaccc
     1981 atacgatgtt cctgactatg cgggctatcc ctatgacgtc ccggactatg caggatccta
     2041 tccatatgac gttccagatt acgcttaaaa gccgaattct gcagatatcc atcacactgg
     2101 cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc
     2161 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2221 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2281 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2341 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2401 ggcttctgag gcggaaagaa ccagctgggg ctcgaggggg gatccccacg cgccctgtag
     2461 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2521 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2581 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2641 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2701 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2761 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2821 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2881 caaaatatta acgcttacaa tttaaatatt tgcttataca atcttcctgt ttttggggct
     2941 tttctgatta tcaaccgggg tgggtaccga gctcgaattc tgtggaatgt gtgtcagtta
     3001 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3061 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3121 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3181 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3241 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3301 ggcctaggct tttgcaaaaa gctcccggga gcttggatat ccattttcgg atctgatcaa
     3361 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3421 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3481 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3541 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3601 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3661 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3721 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3781 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3841 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3901 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3961 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4021 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4081 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4141 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4201 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4261 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4321 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4381 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4441 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgt cgacctcgag
     4501 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4561 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4621 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4681 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4741 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4801 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4861 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4921 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4981 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5041 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5101 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5161 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5221 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5281 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5341 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5401 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5461 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5521 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5581 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5641 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5701 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5761 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5821 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5881 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5941 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6001 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6061 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6121 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6181 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6241 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6301 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6361 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6421 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6481 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6541 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6601 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6661 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     6721 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     6781 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     6841 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     6901 atacgcgtt
//
