LOCUS       sequence-299516-        7603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses kinase-dead Strep-HA-ZAK alpha in mammalian cells, can be 
            used to make inducible cell line.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7603)
  AUTHORS   Vind AC, Snieckute G, Blasius M, Tiedje C, Krogh N, Bekker-Jensen 
            DB, Andersen KL, Nordgaard C, Tollenaere MAX, Lund AH, Olsen JV, 
            Nielsen H, Bekker-Jensen S
  TITLE     ZAKalpha Recognizes Stalled Ribosomes through Partially Redundant 
            Sensor Domains.
  JOURNAL   Mol Cell. 2020 Apr 9. pii: S1097-2765(20)30189-1. doi: 
            10.1016/j.molcel.2020.03.021.
  PUBMED    32289254
REFERENCE   2  (bases 1 to 7603)
  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: "Mol Cell. 
            2020 Apr 9. pii: S1097-2765(20)30189-1. doi: 
            10.1016/j.molcel.2020.03.021."
COMMENT     Sequence Label: pcDNA4/TO/Strep-HA-ZAKalpha_K45A
FEATURES             Location/Qualifiers
     source          1..7603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             993..1016
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1053..1076
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1104..1130
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3611..3628)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3617..3841
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3887..4315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3974..3993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4184..4205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4324..4344)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4329..4658
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4509..4644
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4571..4590
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4706..4753
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4772..5146
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5276..5397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5313..5332)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5367..5386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5446..5462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5446..5462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5459..5481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5470..5486
                     /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(join(5494..5500,5501..5518,5519..5524))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5539..5560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5677..5694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5848..6436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5928..5947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6607..7398,7399..7467))
                     /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     7230..7249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7468..7572)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttatggcta gctggagcca cccgcagttc gagaaaggtg
     1021 gaggttccgg aggtggatcg ggaggtggat cgtggagcca cccgcagttc gaaaaaggcg
     1081 ccgagaccgc ggtcccgaat tcttaccctt atgacgtccc cgattacgcc gctgcagcgg
     1141 atatccagca cagtggcggc cgcatgtcgt ctctcggtgc ctcctttgtg caaattaaat
     1201 ttgatgactt gcagtttttt gaaaactgcg gtggaggaag ttttgggagt gtttatcgag
     1261 ccaaatggat atcacaggac aaggaggtgg ctgtcgcgaa gctcctcaaa atagagaaag
     1321 aggcagaaat actcagtgtc ctcagtcaca gaaacatcat ccagttttat ggagtaattc
     1381 ttgaacctcc caactatggc attgtcacag aatatgcttc tctgggatca ctctatgatt
     1441 acattaacag taacagaagt gaggagatgg atatggatca cattatgacc tgggccactg
     1501 atgtagccaa aggaatgcat tatttacata tggaggctcc tgtcaaggtg attcacagag
     1561 acctcaagtc aagaaacgtt gttatagctg ctgatggagt attgaagatc tgtgactttg
     1621 gtgcctctcg gttccataac catacaacac acatgtcctt ggttggaact ttcccatgga
     1681 tggctccaga agttatccag agtctccctg tgtcagaaac ttgtgacaca tattcctatg
     1741 gtgtggttct ctgggagatg ctaacaaggg aggtcccctt taaaggtttg gaaggattac
     1801 aagtagcttg gcttgtagtg gaaaaaaacg agagattaac cattccaagc agttgcccca
     1861 gaagttttgc tgaactgtta catcagtgtt gggaagctga tgccaagaaa cggccatcat
     1921 tcaagcaaat catttcaatc ctggagtcca tgtcaaatga cacgagcctt cctgacaagt
     1981 gtaactcatt cctacacaac aaggcggagt ggaggtgcga aattgaggca actcttgaga
     2041 ggctaaagaa actagagcgt gatctcagct ttaaggagca ggagcttaaa gaacgagaaa
     2101 gacgtttaaa gatgtgggag caaaagctga cagagcagtc caacaccccg ctgctgcctt
     2161 cctttgagat tggtgcatgg acggaagacg atgtgtattg ttgggttcag cagctcgtca
     2221 gaaaaggtga ctcttcagca gagatgagtg tatatgcaag cttgtttaaa gaaaacaaca
     2281 ttacagggaa gcggctgctg ctgctggagg aagaagacct gaaagacatg ggcattgtct
     2341 ccaaggggca tatcattcac ttcaagtcag ccattgagaa attaacccat gattacataa
     2401 atttgtttca cttcccacca ctaattaagg actcaggagg tgaacctgaa gaaaatgagg
     2461 aaaaaatagt gaacctggaa ctggtttttg gttttcactt gaaaccagga actggcccac
     2521 aggattgtaa gtggaaaatg tatatggaga tggatgggga tgaaattgca ataacctaca
     2581 taaaagatgt gacattcaac actaacctac ctgatgcgga gattttaaag atgacaaagc
     2641 caccatttgt aatggagaag tggattgtag gaatagcaaa aagtcagact gtggagtgca
     2701 ctgtcacata tgagagtgat gttagaactc caaaaagcac taaacatgtc catttgattc
     2761 agtggagtag aacaaaacct caggatgaag tgaaagcagt ccaacttgcc attcagacat
     2821 tattcaccaa ttcagatggc aaccctggaa gcaggtccga ctcaagtgct gattgccagt
     2881 ggttagatac tctgaggatg cggcagattg catccaacac ttctttacag cgttcccaga
     2941 gcaatcctat tctggggtca ccgttcttct cacactttga tggccaggat tcctacgctg
     3001 ctgctgtgag acggccccag gtgcccatta agtatcaaca gattacacct gtgaaccagt
     3061 ccagaagctc gtctcctact cagtatggac tgaccaaaaa cttctcttcc ctacatctca
     3121 actctaggga cagtggcttt tccagtggca atactgacac ctcttcagag aggggtcgat
     3181 actcagacag aagcaggaac aaatatggac gtggtagtat atcactcaat tcttctccta
     3241 gaggaagata cagtggaaag agtcagcatt ccactccttc aagaggaaga taccctggaa
     3301 agttctacag ggtttctcag tcagcactca atcctcacca gtcgcctgac ttcaagagaa
     3361 gccccaggga cctccaccaa cccaacacca taccagggat gcctttgcac cctgagactg
     3421 actcaagagc cagtgaagag gacagcaaag tcagcgaagg gggctggaca aaagtggaat
     3481 accggaaaaa gccccacagg ccatctcccg ccaaaaccaa taaagagaga gccagagggg
     3541 accaccgtgg atggagaaac ttttgagcgg ccgctcgagt ctagagggcc cgtttaaacc
     3601 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3661 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3721 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3781 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3841 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3901 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3961 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4021 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4081 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4141 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4201 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4261 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4321 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4381 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4441 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4501 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4561 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4621 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4681 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
     4741 agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt
     4801 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
     4861 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
     4921 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
     4981 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
     5041 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
     5101 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga
     5161 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5221 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5281 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5341 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5401 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5461 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5521 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5581 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5641 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5701 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5761 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5821 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5881 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5941 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6001 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6061 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6121 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6181 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6241 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6301 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6361 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6421 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6481 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6541 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6601 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6661 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6721 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6781 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6841 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6901 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6961 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7021 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7081 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7141 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7201 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7261 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7321 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7381 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7441 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7501 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7561 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
