LOCUS       sequence-366765-        5447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the C. elegans protein H9G2U1_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 5447)
  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 5447)
  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_H9G2U1_CAEEL (pBS0346)
FEATURES             Location/Qualifiers
     source          1..5447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(58..77)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        249..628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        629..832
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     783..803
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     877..896
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        877..895
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1429..1452
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1469..1486)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1475..1699
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1745..2173
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1832..1851)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2042..2063
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2182..2202)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2187..2516
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2367..2502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2429..2448
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2564..2611
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2630..3004
                     /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    3134..3255
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3171..3190)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3225..3244
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3304..3320)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3304..3320)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3317..3339)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3328..3344
                     /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(3352..3358,3359..3376,3377..3382))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3535..3552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3706..4294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3786..3805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4465..5256,5257..5325))
                     /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     5088..5107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5326..5430)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
       61 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
      121 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
      181 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
      241 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      301 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      361 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      421 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      481 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      541 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      601 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      661 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      721 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      781 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
      841 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      901 ccaagctggc tagccaccat gtatgatgca gctgcagaag gagcaagtaa tatggcacag
      961 tccatgcacg aggctggaaa atccaccggc gacgcccttt cagagggtgc cgaagctggc
     1021 tacgatgcgg taaaggacgg aggcaggtgg gtgggagaaa acgtcgccca tggagcaggt
     1081 aaagtggccg acggagccaa ggctgcagga ggcgccgtgg ttgatggagt agccgcagca
     1141 ggaggagctg ttgttggtgg agctaaggct gtgggatcag ggattgctga tggggctaag
     1201 tttgtggggg aaaacgtggc atatggtgcc ggagcagtgg caggaggagc aaaggcagca
     1261 ggaggggcag tggtggacgg tgccgcagcc gctggaggag ccgtggtgga cggagccaaa
     1321 gctgtgggag gcttcgtgag tgatacattt tcttctgcca gagacgagac aggagaggcc
     1381 ttgaagacag ctggcagcaa agtgaaggcc tctgagtgag gatctgggga ctacaaggac
     1441 gacgacgaca agtgaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     1501 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1561 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1621 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     1681 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     1741 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1801 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1861 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1921 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1981 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2041 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2101 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2161 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2221 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2281 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2341 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2401 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     2461 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2521 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     2581 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
     2641 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
     2701 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
     2761 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
     2821 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
     2881 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
     2941 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
     3001 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3061 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3121 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3181 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3241 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     3301 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3361 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac ttacattaat
     3421 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3481 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3541 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3601 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3661 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3721 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3781 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3841 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3901 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3961 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4021 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4081 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4141 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4201 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4261 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4321 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4381 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4441 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4501 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4561 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4621 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4681 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4741 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4801 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4861 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4921 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4981 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5041 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5101 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5161 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5221 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5281 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5341 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5401 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaag
//
