LOCUS       sequence-366744-        7148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the C. elegans protein G5EFU3_CAEEL.
            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 7148)
  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 7148)
  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_G5EFU3_CAEEL (pBS0308)
FEATURES             Location/Qualifiers
     source          1..7148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        233..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        613..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     767..787
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     861..880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        861..879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3114..3137
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3154..3171)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3160..3384
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3430..3858
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3517..3536)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3727..3748
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3867..3887)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3872..4201
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4052..4187
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4114..4133
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4249..4296
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4315..4689
                     /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    4819..4940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4856..4875)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4910..4929
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4989..5005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4989..5005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5002..5024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5013..5029
                     /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(5037..5043,5044..5061,5062..5067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5220..5237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5391..5979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5471..5490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6150..6941,6942..7010))
                     /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     6773..6792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7011..7115)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
       61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagcca
      901 ccatgtcatt catggataag gtcaaagata cagctaattc cgcctacgag tcagtcgaaa
      961 acacagccgt cggtgctaaa gatggtgtcg tgggcgcttt caatactgtc ggtgagaaaa
     1021 ttggtggcgc ctacgatagt acgaagcaat atgctagcga taaggttagt gcggccgggg
     1081 attatgctag cgaagcaggt aagaagaagg tggatgcgga agactctatg atccaagggg
     1141 cttatgatgc taaagacagt gctaaagagt ctgcgtcaaa tgcgggcaat aaaatttcta
     1201 atgccttcac ttcccttaaa gatagtataa ctggtgacaa tgctcacaaa actggcgaag
     1261 ctcttaacga taactatgaa agcgttaaag aaagtgcatc tgaggcagcc gatcgcctca
     1321 aggaacccgc ccaaaacacg gcagataatg caaaggatct tgcaaaccag acaggggaag
     1381 agatgtcaga tgcatttgag agcgtcaaac aattgacttc cgatacagct gaaaatgcca
     1441 aagagaatgc tcatgatgca gcggataccg caaaatcata caaggacaaa gcaggagaaa
     1501 aggcgtccgg tattttggac tcctttaaag cgcatgcatc cgatgcaaag gattccgttg
     1561 agaacaaagc ctcagatgcc tataatagtg caaaagataa agcaggcgat gcttgggatt
     1621 ctactaaaga tgcagcggga gatgcaaaag ataaaacaaa gagtttcacg gagaaagtcg
     1681 gcgataaaat atccggagcc tacgattcag taaaagagaa ggcctcagat gtcgcagata
     1741 gcttcaaagc ccacagtact gatagcaaag ataatgtaga aaacaaagct gccgatgcat
     1801 ataacacagc caaagataaa gcctcagatg catgggacaa aacaaaagat aaggctggtg
     1861 aggcgaaaga caagatggga gatgcttggg ataccaccaa agacaaagct ggcgatgcct
     1921 gggatacgac aaaggataag gccggtgatg gtaaggggaa agccggggat gcctgggata
     1981 caacaaagga taaagcctct gatgcttggg atacgacgaa agataaagca ggtgaagcga
     2041 aggacaaaat gggcgaagca tgggatcata caaaagacaa agcaggggaa gctaaggaca
     2101 aggcatccga cgcagctgat gatgcgcaag gaaagtctaa gtccatgacc gaaaagattg
     2161 gcgattccat tagcggagcc tgggaatcca ccaaagacac ggcagtctcc gcaaaagata
     2221 aaactgccga agcagctgga aaggttggag actcaatcag cggggcttat gataccgtga
     2281 aggaaaaggc aagtgacatt gcggacagtt ttaaagcgca ctccaccgac tccaaggata
     2341 atgtcgaaaa caaagccgct gatgcctata acagcgccaa ggataaagca agcgatgctt
     2401 gggataaaac aaaagataag gcaggggatg cgaaagataa agccgccgat gcgtgggata
     2461 cgactaaaga taaagcagga gatgcgtggg attccactaa agaccatgct gccgatgcta
     2521 aagataaagc tagcgatgca gctggggatg cgaaagataa aagtaaatca ttgaccgaaa
     2581 aggcaggcga tgccataagt ggcgcttatg actccgtaaa agagaaggct tctgatatag
     2641 cagactcctt taaagcgcat tccactaatt ccaaagataa tgttgagaat aaagctagtg
     2701 atgcatataa ttctgcgaaa gacaaggctt ctgacgcttg ggataaaact aaggataaag
     2761 ccggcgaagc aaaagacaaa gcgggagatg cttgggataa cacaaaagat aaagcgggaa
     2821 atgcctggga tagcacgaaa gacaaggcat cagatgcttg ggatacaacg aaagataaag
     2881 cttccgatgc taaggaatca gctggggatg ctgcagattc tgcaaaagac aagtcaaaat
     2941 ccatcacaga aactatcggc gataaaatct ccggcgcata tgacagcgtt aaagagaaag
     3001 cttccgatgt tgctgactcc tttaaagctc attcttcaga tgcacaagat actgtggaag
     3061 tttctcacca taagtatttc aatgttttct catcttttga ataaggatct ggggactaca
     3121 aggacgacga cgacaagtga acccgctgat cagcctcgac tgtgccttct agttgccagc
     3181 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3241 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3301 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3361 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3421 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3481 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3541 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3601 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3661 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3721 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3781 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3841 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3901 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3961 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4021 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4081 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4141 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4201 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
     4261 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     4321 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
     4381 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
     4441 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
     4501 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
     4561 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
     4621 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
     4681 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4741 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4801 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4861 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4921 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4981 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5041 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaacttaca
     5101 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5161 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5221 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5281 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5341 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5401 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5461 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5521 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5581 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5641 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5701 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5761 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5821 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5881 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5941 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6001 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6061 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6121 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6181 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6241 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6301 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6361 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6421 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6481 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6541 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6601 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6661 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6721 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6781 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6841 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6901 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6961 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7021 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7081 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7141 gacgtcga
//
