LOCUS       sequence-366740-        5960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the Deinococcus radiodurans protein 
            DR_B0118. 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 5960)
  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 5960)
  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_DR_B0118 (pBS0303)
FEATURES             Location/Qualifiers
     source          1..5960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..85)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        257..636
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        637..840
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     791..811
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     885..904
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        885..903
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1950..1973
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1990..2007)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1996..2220
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2266..2694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2353..2372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2563..2584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2703..2723)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2708..3037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2888..3023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2950..2969
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3085..3132
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3151..3525
                     /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    3655..3776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3692..3711)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3746..3765
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3825..3841)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3825..3841)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3838..3860)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3849..3865
                     /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(3873..3879,3880..3897,3898..3903))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4056..4073)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4227..4815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4307..4326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4986..5777,5778..5846))
                     /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     5609..5628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5847..5951)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
       61 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
      121 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
      181 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      241 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      301 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      361 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      421 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      481 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      541 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      601 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      661 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      721 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      781 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      841 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      901 agggagaccc aagctggcta gccaccatga gcactccggt ggagccactt ctctgcctga
      961 gtagcgtatg tgacactcgt aaagctatga aggaggagat gcagtcaacc cgcaggcgct
     1021 tcctgggtat ggccggcgct atgggcgcag gggcagtcct ggctggatgc gccaatgtgg
     1081 gcgccagcga gccgacaaaa accaacctgg atgccacaat ctttaacttc gccttgaacc
     1141 tggaatacct ggaggcagct ttctatctgg ccgccgttgg ccggcttaat gagcttaccg
     1201 ccgctggggg tgacgcatcc aaggtcactc taccttccgg cgtgaccggt atgggaggca
     1261 cggctgtgcc tgggctgacc ggcgatctgc gcgctatgat ggaggagatt gccgatgacg
     1321 agctcgccca cgtgaaagtc atccggtccg tgctgggtag tgccgccgtg gcccagccta
     1381 ggctggattt atccgccagc tttctggccg ccgggtcctt ggccagtaac ggtgcaatca
     1441 ccaacttcaa cccctacgct aaccccctgt ttttcttgca cggggctttc gtctttgagg
     1501 acgtcggcgt gactgcctac aaaggggctg cacggcttct ggtgggcgat aagcctggag
     1561 ggaatctgga gaacgcagcc ggaatcctgg ccgtggaagc gtaccacgcc gggagcatcc
     1621 ggacccagct ctttatgcga cgcaccgagc aggccgccgc cggcctaaca gtggagcaag
     1681 tggtgcaggc tatttccaat cttagggaca gcgtcgatgg cgccgacgac agggaccagg
     1741 gcatcactgc aaatggcaac gctggggttt tggctcgaga cgctaacatc attccaaccg
     1801 actctaacgg aattgccttc tccagaactc ccagacaggt cgctaacatc gtcttcctgg
     1861 acacaaccgg caaggccgca aggggaggct tcttcccaga cgggctgacc ggggactact
     1921 cttctatttt atctttataa ggatctgggg actacaagga cgacgacgac aagtgaaccc
     1981 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2041 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2101 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2161 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2221 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     2281 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2341 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2401 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2461 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2521 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2581 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2641 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     2701 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2761 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2821 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2881 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2941 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3001 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     3061 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     3121 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     3181 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     3241 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     3301 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     3361 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     3421 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     3481 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
     3541 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3601 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3661 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3721 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3781 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3841 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3901 aaagcctggg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     3961 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4021 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4081 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4141 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4201 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4261 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4321 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4381 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4441 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4501 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4561 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4621 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4681 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4741 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4801 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4861 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4921 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4981 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5041 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5101 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5161 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5221 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5281 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5341 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5401 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5461 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5521 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5581 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5641 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5701 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5761 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5821 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5881 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5941 ggggttccgc gcacatttcc
//
