LOCUS       sequence-363782-        5594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the durm wheat protein DHN_TaDHN5. 
            This is one of the bottom performing targets in our apoptosis assay 
            (it induced apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5594)
  AUTHORS   Veling MT, Nguyen DT, Thadani NN, Oster ME, Rollins NJ, Brock KP, 
            Bethel NP, Lim S, Baker D, Way JC, Marks DS, Chang RL, Silver PA
  TITLE     Natural and Designed Proteins Inspired by Extremotolerant Organisms 
            Can Form Condensates and Attenuate Apoptosis in Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Mar 18;11(3):1292-1302. doi: 
            10.1021/acssynbio.1c00572. Epub 2022 Feb 18.
  PUBMED    35176859
REFERENCE   2  (bases 1 to 5594)
  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"; doi: 
            "10.1021/acssynbio.1c00572"; journalName: "ACS Synth Biol"; date: 
            "2022-03-18- 18"; volume: "11"; issue: "3"; pages: "1292-1302"
COMMENT     Sequence Label: pST_DHN_TaDHN5 (pBS0801)
FEATURES             Location/Qualifiers
     source          1..5594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(36..55)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        227..606
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        607..810
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     761..781
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     855..874
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        855..873
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      891..900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1594..1611)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1600..1824
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1870..2298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1957..1976)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2167..2188
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2307..2327)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2312..2641
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2492..2627
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2554..2573
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2689..2736
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2755..3129
                     /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    3259..3380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3296..3315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3350..3369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3429..3445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3429..3445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3442..3464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3453..3469
                     /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(3477..3483,3484..3501,3502..3507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3660..3677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3831..4419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3911..3930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4590..5381,5382..5450))
                     /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     5213..5232
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5451..5555)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
       61 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
      121 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
      181 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
      241 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      301 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      361 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      421 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      481 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      541 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      601 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      661 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      721 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      781 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      841 tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta gccaccatgg
      901 aatttcaagg acaacatgat aatcctgcta atagagtaga tgaatatgga aatccatttc
      961 cgttggcagg cggagtcggc ggtgcccacg cggccccagg cacaggtgga caatttcaag
     1021 cacatcgggg agaacataaa acaggtggta tactccatcg atccgggagt agtagttctt
     1081 caagttcatc tgaagatgat ggaatgggcg gccgaagaaa gaaaggtatg aaagaaaaga
     1141 ttaaagaaaa gcttcctggc gggcatgggg atcaacaaca cacgggcggc acttacagac
     1201 aacaaggaac tggtatggtt ggtactggcg gtacttatgg gcagaaaggg catactggga
     1261 tgacagggac aggtggaacg tatggtcaac agggtcatac tggcatgaca ggaacaggcg
     1321 gaacgtatgg acaacaaggc cacacaggta tgacaggcac gggtggaaca tatgggcaac
     1381 aaggtcacac aggtatggcc ggtacagggg ctcatggaac tactgcaaca ggtggtacgt
     1441 atggccaacc gggccatacg ggaatgacag gaacaggggc acacgggaca ggcggtacat
     1501 atgggcaaca tggcacggat actggagaaa agaaaggaat aatggataaa attaaagaga
     1561 aactcccggg tcaacattga acccgctgat cagcctcgac tgtgccttct agttgccagc
     1621 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1681 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     1741 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     1801 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     1861 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1921 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1981 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2041 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2101 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2161 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2221 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2281 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2341 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2401 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2461 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2521 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2581 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2641 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
     2701 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     2761 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
     2821 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
     2881 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
     2941 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
     3001 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
     3061 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
     3121 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3181 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     3241 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3301 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3361 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     3421 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3481 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaacttaca
     3541 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3601 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3661 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3721 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3781 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3841 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3901 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3961 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4021 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4081 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4141 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4201 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4261 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4321 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4381 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4441 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4501 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4561 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4621 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4681 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4741 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4801 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4861 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4921 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4981 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5041 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5101 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5161 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5221 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5281 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5341 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5401 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5461 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5521 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5581 gacgtcgacg gatc
//
