LOCUS       sequence-363784-        5714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the Eutrema salsugineum protein 
            DHN_TsDHN-1. This is one of the top performing targets in our 
            apoptosis assay (it protected from apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5714)
  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 5714)
  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_TsDHN-1 (pBS0805)
FEATURES             Location/Qualifiers
     source          1..5714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..830
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     875..894
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        875..893
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      911..920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1734..1751)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1740..1964
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2010..2438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2097..2116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2307..2328
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2447..2467)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2452..2781
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2632..2767
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2694..2713
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2829..2876
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2895..3269
                     /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    3399..3520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3436..3455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3490..3509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3569..3585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3569..3585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3582..3604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3593..3609
                     /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(3617..3623,3624..3641,3642..3647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3800..3817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3971..4559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4051..4070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4730..5521,5522..5590))
                     /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     5353..5372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5591..5695)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      841 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      901 aagctggcta gccaccatgg ccgaggagta caagaacgcc agcgaggagt tcaagaacgt
      961 gcccgagcac gagacaaccc ccaagatcag caccaccgag gagcccagcg ccgaggtgaa
     1021 ggacaggggc ttcttcgact tcctgggcaa gaagaaggag gaggtgaagc cccaggagac
     1081 aaccaccccc ctggagagcg agttcgagca caaggcccag atcagcgagc cccccgcctt
     1141 cgtggccaag catgaggagg agcaggagac aaaggagaac aagcccaccc tggtggagca
     1201 gctgcaccag aagcacgtgg aggaggagga gaacaagccc agcctgttcg acaagctgca
     1261 caggtcaagt tccagttcct catcatcatc agacgaggag ggcgaggacg gagagaagag
     1321 gaagaagaag aaggagaaga agaagaccgt ggagggcgag gacaagaccg aggaggagaa
     1381 caagggcgtg atggacaaga tcaaggagaa gttcccccac gccaagaaga ccgaggacga
     1441 ccacgccccc gtggttaccg gagtgcccga gacagagaag atcggcatga ccgagaaaat
     1501 aaaagagaaa ttgccaggac atggcaagaa gcccgaggac agccccgtgg ttgataccgc
     1561 tcccgtggtg gagacagcca cccccatcac cgccgagcac tctgctgagc accccgctga
     1621 gaacaagggc ttcctggaga aaattaaaga gaaattgcct ggtcatcacg ccaagggcac
     1681 cgaggagatg gagaagaagg agaaggagag cgacgcctga acccgctgat cagcctcgac
     1741 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1801 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1861 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1921 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     1981 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2041 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2101 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2161 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2221 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2281 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2341 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2401 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2461 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2521 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2581 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2641 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2701 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     2761 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     2821 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     2881 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     2941 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     3001 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     3061 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     3121 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     3181 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     3241 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     3301 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3361 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3421 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3481 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     3541 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3601 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3661 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3721 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3781 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3841 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3901 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3961 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4021 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4081 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4141 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4201 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4261 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4321 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4381 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4441 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4501 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4561 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4621 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4681 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4741 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4801 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4861 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4921 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4981 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5041 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5101 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5161 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5221 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5281 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5341 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5401 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5461 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5521 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5581 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5641 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5701 ttccccgaaa agtg
//
