LOCUS       sequence-366762-        5750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the C. elegans protein H9G2T8_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 5750)
  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 5750)
  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_H9G2T8_CAEEL (pBS0342)
FEATURES             Location/Qualifiers
     source          1..5750
                     /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             1716..1739
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1756..1773)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1762..1986
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2032..2460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2119..2138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2329..2350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2469..2489)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2474..2803
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2654..2789
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2716..2735
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2851..2898
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2917..3291
                     /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    3421..3542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3458..3477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3512..3531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3591..3607)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3591..3607)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3604..3626)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3615..3631
                     /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(3639..3645,3646..3663,3664..3669))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3822..3839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3993..4581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4073..4092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4752..5543,5544..5612))
                     /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     5375..5394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5613..5717)
                     /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 ccatgcgcat ggtaggcgag aacgtcgccc atggagccgg gaaggtggcg gacggggcca
      961 aggccgccgg caattacgcc gtagatagcg ccaacaccgc cggcgaggcc attgtggacg
     1021 gcgcaaagac ggtgggccag acagcttccg atgggatgcg catgatcggc gagaacgtgg
     1081 cccacggggc cggagtggtg gtggagggag ctaagtctgc tggacaatct attgctgatg
     1141 gagctgtagc agctaaggac gctgcagtgg agggagctac tactgcaggt aacaaaacag
     1201 tggaaggtat gaggtttgtg ggagaaaacg tggcacatgg agctgggaaa gtggcagatg
     1261 gtgcaaaagc agcaggggat acagttgtag aagggggtaa agctgccgct gaaaagaccg
     1321 aatctggcat tgcatttgta ggacagaaaa ttgcagaagg agccggagca gtggcaggtg
     1381 gagctaaagc tgctggaggg gctgtggtgg atggggtggc tgctgctgga ggggctgtcg
     1441 tgggtggagc taaagctgta ggaagcggta tcgctgatgg ggctaaattt gttggtgaaa
     1501 acgttgctta tggagctgga gctgttgcag ggggagccaa ggcagccgga ggagccgttg
     1561 ttgatggcgc tgcagctgcc ggtggtgcag tggtggatgg tgctaaggca gtgggaggct
     1621 ttgtatctga tacattttct agtgcaaggg acgaaacagg tgaggcctta aaaactgctg
     1681 gaagtaaggt gaaggccagt gagtgaggat ctggggacta caaggacgac gacgacaagt
     1741 gaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1801 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1861 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1921 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     1981 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2041 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2101 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2161 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2221 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2281 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2341 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2401 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2461 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2521 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2581 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2641 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2701 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2761 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     2821 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     2881 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
     2941 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     3001 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     3061 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     3121 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     3181 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     3241 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
     3301 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3361 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     3421 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3481 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3541 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     3601 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3661 aagtgtaaag cctggggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     3721 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3781 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3841 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3901 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3961 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4021 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4081 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4141 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4201 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4261 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4321 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4381 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4441 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4501 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4561 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4621 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4681 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4741 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4801 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4861 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4921 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4981 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5041 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5101 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5161 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5221 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5281 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5341 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5401 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5461 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5521 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5581 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5641 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5701 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
//
