LOCUS       sequence-300399-        7393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Strep-HA-ZAKalpha_deltaSdeltaCTD in mammalian cells, can 
            be used to make inducible cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7393)
  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 7393)
  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_deltaSdeltaCTD
FEATURES             Location/Qualifiers
     source          1..7393
                     /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(3401..3418)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3407..3631
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3677..4105
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3764..3783)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3974..3995
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4114..4134)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4119..4448
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4299..4434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4361..4380
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4496..4543
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4562..4936
                     /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    5066..5187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5103..5122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5157..5176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5236..5252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5236..5252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5249..5271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5260..5276
                     /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(5284..5290,5291..5308,5309..5314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5329..5350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5467..5484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5638..6226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5718..5737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6397..7188,7189..7257))
                     /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     7020..7039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7258..7362)
                     /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 ctgtaaagaa 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 aagttctaca
     3181 gggtttctca gtcagcactc aatcctcacc agtcgcctga cttcaagaga agccccaggg
     3241 acctccacca acccaacacc ataccaggga tgcctttgca ccctgagact gactcaagag
     3301 ccagtgaaga ggacagcaaa gtcagcgaag ggggctggac aaaagtggaa tactaagcgg
     3361 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3421 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3481 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3541 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3601 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3661 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3721 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3781 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3841 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3901 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3961 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4021 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4081 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4141 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4201 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4261 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4321 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4381 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4441 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     4501 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     4561 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     4621 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     4681 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     4741 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     4801 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     4861 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     4921 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     4981 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5041 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5101 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5161 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5221 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5281 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5341 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5401 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5461 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5521 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5581 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5641 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5701 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5761 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5821 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5881 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5941 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6001 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6061 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6121 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6181 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6241 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6301 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6361 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6421 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6481 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6541 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6601 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6661 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6721 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6781 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6841 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6901 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6961 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7021 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7081 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7141 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7201 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7261 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7321 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7381 gccacctgac gtc
//
