LOCUS       sequence-299550-        7474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Strep-HA-ZAKalpha_deltaS 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 7474)
  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 7474)
  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_deltaS
FEATURES             Location/Qualifiers
     source          1..7474
                     /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(3482..3499)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3488..3712
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3758..4186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3845..3864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4055..4076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4195..4215)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4200..4529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4380..4515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4442..4461
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4577..4624
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4643..5017
                     /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    5147..5268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5184..5203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5238..5257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5317..5333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5317..5333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5330..5352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5341..5357
                     /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(5365..5371,5372..5389,5390..5395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5410..5431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5548..5565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5719..6307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5799..5818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6478..7269,7270..7338))
                     /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     7101..7120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7339..7443)
                     /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 taccggaaaa
     3361 agtcccacag gccatctccc gccaaaacca ataaagagag agccagaggg gaccaccgtg
     3421 gatggagaaa cttttgagcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     3481 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3541 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3601 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3661 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3721 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3781 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3841 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3901 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3961 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4021 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4081 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4141 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4201 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4261 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4321 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4381 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4441 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4501 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4561 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     4621 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     4681 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     4741 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     4801 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     4861 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     4921 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     4981 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     5041 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5101 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5161 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5221 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5281 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5341 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5401 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5461 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5521 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5581 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5641 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5881 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6121 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7441 gcgcacattt ccccgaaaag tgccacctga cgtc
//
