LOCUS       sequence-366763-        5567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the C. elegans protein H9G2T9_CAEEL.
            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 5567)
  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 5567)
  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_H9G2T9_CAEEL (pBS0344)
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     862..881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        862..880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1534..1557
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1574..1591)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1580..1804
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1850..2278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1937..1956)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2147..2168
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2287..2307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2292..2621
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2472..2607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2534..2553
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2669..2716
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2735..3109
                     /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    3239..3360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3276..3295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3330..3349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3409..3425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3409..3425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3422..3444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3433..3449
                     /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(3457..3463,3464..3481,3482..3487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3640..3657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3811..4399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3891..3910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4570..5361,5362..5430))
                     /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     5193..5212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5431..5535)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      841 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcc
      901 accatgtatg atgccgcggc ggagggggcc agtaacatgg cccagagtat gcacgaggcc
      961 ggcaagagca ccggcgacgc actgtccgag ggcgccgagg ctggctacga cgcagtgaag
     1021 gacgggggca gatgggtggg cgagaacgtg gcccacggcg ccggcaaggt ggccgacggc
     1081 gcaaaggccg ctggtgacac cgtggtggag ggagggaaag ccgcagcaga aaaaactgaa
     1141 agcggtatag catttgtagg acagaagatc gcagaggggg ctggcgccgt ggccggcggc
     1201 gccaaagctg ccggaggagc cgtggtggat ggcgtggctg ccgctggggg tgcagtggtc
     1261 gtgggtgcaa aagcagtagg atcaggaata gcagatgggg caaaatttgt gggggaaaac
     1321 gtggcatatg gggctggggc tgtggcagga ggagcaaaag ctgcaggtgg tgcagtggtg
     1381 gatggagcag ctgcagcagg aggagcagtt gttgatggag caaaagcagt tggaggattt
     1441 gtttctgaca ctttttctag tgcaagagac gaaactggag aagcactgaa gactgcagga
     1501 tcaaaagtta aagcctcaga ataaggatct ggggactaca aggacgacga cgacaagtga
     1561 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1621 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     1681 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     1741 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     1801 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     1861 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1921 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1981 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2041 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2101 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2161 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2221 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     2281 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2341 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2401 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2461 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2521 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     2581 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     2641 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     2701 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
     2761 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     2821 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     2881 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     2941 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     3001 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     3061 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg
     3121 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3181 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     3241 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3301 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3361 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     3421 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3481 gtgtaaagcc tggggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     3541 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3601 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3661 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3721 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3781 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3841 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3901 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3961 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4021 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4081 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4141 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4201 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4261 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4321 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4381 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4441 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4501 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4561 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4621 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4681 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4741 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4801 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4861 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4921 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4981 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5041 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5101 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5161 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5221 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5281 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5341 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5401 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5461 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5521 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcg
//
