LOCUS       sequence-366841-        5303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the Deinococcus radiodurans protein 
            Q96Q06-2_DrHD_b03_dom3. 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 5303)
  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 5303)
  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_Q96Q06-2_DrHD_b03_dom3 (pBS0687)
FEATURES             Location/Qualifiers
     source          1..5303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..82)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        254..633
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        634..837
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     788..808
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     882..901
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        882..900
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      918..927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1330..1347)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1336..1560
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1606..2034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1693..1712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1903..1924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2043..2063)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2048..2377
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2228..2363
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2290..2309
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2425..2472
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2491..2865
                     /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    2995..3116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3032..3051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3086..3105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3165..3181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3165..3181)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3178..3200)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3189..3205
                     /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(3213..3219,3220..3237,3238..3243))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3396..3413)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3567..4155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3647..3666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4326..5117,5118..5186))
                     /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     4949..4968
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5187..5291)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
       61 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
      121 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
      181 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
      241 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      301 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      361 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      421 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      481 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      541 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      601 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      661 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      721 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      781 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      841 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
      901 gagacccaag ctggctagcc accatgggag tggacactac caagatcgtg ctgacaggca
      961 ccaaggacac agtgtgctct ggagtgacag gcgccgccaa tgtggccaag ggctgatgaa
     1021 ttgccaattg aaatttgacg gcctatgtca ctgaaactat ccaaacaccc catgtcgata
     1081 ctgaacgtat cgacgcatac ctccttcctt gaaaacgcac aatcatacaa ctgggcacat
     1141 aatgcgtacg cccatctagt acacccatct ctgtaggtcc agttcaagag ctggaagagc
     1201 accctccact tagacgtgcc gtaagataga ggttagtgag aatagccatc caatatgagc
     1261 gttgatagct agacgaggat aaggcagtga tagaaggggc accaagcttc attcgcaccc
     1321 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     1381 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     1441 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     1501 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     1561 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     1621 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1681 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1741 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1801 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1861 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1921 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1981 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     2041 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2101 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2161 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2221 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2281 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     2341 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     2401 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     2461 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     2521 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     2581 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     2641 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     2701 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     2761 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     2821 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
     2881 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     2941 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3001 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3061 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3121 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     3181 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3241 aaagcctggg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     3301 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3361 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3421 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3481 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3541 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3601 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3661 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3721 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3781 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3841 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3901 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3961 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4021 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4081 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4141 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4201 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4261 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4321 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4381 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4441 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4501 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4561 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4621 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4681 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4741 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4801 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4861 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4921 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4981 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5041 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5101 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5161 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5221 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5281 ggggttccgc gcacatttcc ccg
//
