LOCUS       sequence-299546-        7522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Strep-HA-ZAKalpha_deltaCTD 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 7522)
  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 7522)
  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_deltaCTD
FEATURES             Location/Qualifiers
     source          1..7522
                     /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(3530..3547)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3536..3760
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3806..4234
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3893..3912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4103..4124
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4243..4263)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4248..4577
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4428..4563
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4490..4509
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4625..4672
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4691..5065
                     /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    5195..5316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5232..5251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5286..5305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5365..5381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5365..5381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5378..5400)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5389..5405
                     /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(5413..5419,5420..5437,5438..5443))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5458..5479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5596..5613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5767..6355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5847..5866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6526..7317,7318..7386))
                     /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     7149..7168
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7387..7491)
                     /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 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 actaagcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     3541 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3601 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3661 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3721 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3781 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3841 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3901 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3961 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4021 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4081 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4141 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4201 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4261 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4321 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4381 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4441 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4501 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4561 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     4621 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     4681 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
     4741 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
     4801 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
     4861 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
     4921 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
     4981 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
     5041 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     5101 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5161 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5221 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5281 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5341 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5401 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5461 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5521 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7021 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7081 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7141 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7201 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7261 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7321 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7381 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7501 ccgaaaagtg ccacctgacg tc
//
