LOCUS       sequence-363746-        5303 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For the mammalian expression of the Deinococcus radiodurans protein 
            DrHD_dom3_k3_7. 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 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 Apr 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_DrHD_dom3_k3_7 (pBS0755)
FEATURES             Location/Qualifiers
     source          1..5303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     866..885
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        866..884
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      902..911
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      906..915
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1314..1331)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1320..1544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1590..2018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1677..1696)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1887..1908
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2027..2047)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2032..2361
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2212..2347
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2274..2293
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2409..2456
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2475..2849
                     /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    2979..3100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3016..3035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3070..3089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3149..3165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3149..3165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3162..3184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3173..3189
                     /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(3197..3203,3204..3221,3222..3227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3380..3397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3551..4139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3631..3650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4310..5101,5102..5170))
                     /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     4933..4952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5171..5275)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      841 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      901 agccaccatg gcggctagcg atgctaaaga taaagctcaa gatgttaagg cagacgcgag
      961 cgacgcagcc gacgacgcga aagacaaagc ccaagattga tgaattgcca attgaaattt
     1021 gacggcctat gtcactgaaa ctatccaaac accccatgtc gatactgaac gtatcgacgc
     1081 atacctcctt ccttgaaaac gcacaatcat acaactgggc acataatgcg tacgcccatc
     1141 tagtacaccc atctctgtag gtccagttca agagctggaa gagcaccctc cacttagacg
     1201 tgccgtaaga tagaggttag tgagaatagc catccaatat gagcgttgat agctagacga
     1261 ggataaggca gtgatagaag gggcaccaag cttcattcgc acccgctgat cagcctcgac
     1321 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1381 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1441 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1501 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     1561 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     1621 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1681 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1741 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1801 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1861 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1921 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1981 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2041 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2101 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2161 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2221 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2281 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     2341 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     2401 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     2461 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     2521 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     2581 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     2641 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     2701 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     2761 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     2821 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     2881 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     2941 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3001 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3061 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     3121 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3181 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3241 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3301 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3361 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3421 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3481 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3541 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3601 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3661 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3721 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3781 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3841 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3901 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3961 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4021 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4081 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4141 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4201 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4261 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4321 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4381 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4441 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4501 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4561 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4621 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4681 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4741 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4801 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4861 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4921 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4981 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5041 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5101 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5161 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5221 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5281 ttccccgaaa agtgccacct gac
//
