LOCUS       sequence-366741-        5282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the resurrection plant 
            (Craterostigma plantagineum) protein Q3YMR1_CRAPL. 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 5282)
  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 5282)
  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_Q3YMR1_CRAPL (pBS0304)
FEATURES             Location/Qualifiers
     source          1..5282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..54)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        226..605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        606..809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     760..780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     854..873
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        854..872
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1241..1264
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1281..1298)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1287..1511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1557..1985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1644..1663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1854..1875
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1994..2014)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1999..2328
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2179..2314
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2376..2423
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2442..2816
                     /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    2946..3067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2983..3002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3037..3056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3116..3132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3116..3132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3129..3151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3140..3156
                     /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(3164..3170,3171..3188,3189..3194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3347..3364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3518..4106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3598..3617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4277..5068,5069..5137))
                     /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     4900..4919
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5138..5242)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
       61 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
      121 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
      181 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
      241 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      301 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      361 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      421 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      481 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      541 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      601 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      661 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      721 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      781 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
      841 ggcttatcga aattaatacg actcactata gggagaccca agctggctag ccaccatgca
      901 gaccgccaag cagaaagtgt ctgacgccgc cgcctctgcc cagcagaacg tggataagat
      961 cagagctaaa gcccaggaga aggctgacaa gacagatgcc agaacctctg aggagaagga
     1021 gatggcccat gagagaaagg aaatttctga ggccgaggct aagatgaaga agcaccagtc
     1081 taaagcccag aacgtggaca aaaaagccca cagccaccag cagccccctt tgggagctgc
     1141 cgctccccct acagatggag gagtgacacc tacacctagc cctgctgccg gcggacagta
     1201 ccctggcgga gccggctcac atgtgatgta aggatctggg gactacaagg acgacgacga
     1261 caagtgaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1321 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1381 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1441 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1501 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1561 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1621 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1681 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1741 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1801 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1861 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1921 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1981 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2041 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2101 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2161 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2221 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2281 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2341 gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt
     2401 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
     2461 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     2521 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     2581 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     2641 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
     2701 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
     2761 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
     2821 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     2881 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     2941 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3001 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3061 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     3121 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3181 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta acttacatta attgcgttgc
     3241 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3301 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3361 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3421 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3481 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3541 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3601 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3661 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3721 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3781 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3841 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3901 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3961 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4021 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4081 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4141 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4201 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4261 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4321 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4381 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4441 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4501 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4561 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4621 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4681 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4741 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4801 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4861 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4921 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4981 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5041 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5101 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5161 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5221 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     5281 cg
//
