LOCUS       sequence-366856-        5282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the sleeping chironomid protein 
            pvLEA22mer_mutant_23. 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_pvLEA22mer_mutant_23 (pBS0719)
FEATURES             Location/Qualifiers
     source          1..5282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..58)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        230..609
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        610..813
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     764..784
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     858..877
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        858..876
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      894..903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1285..1302)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1291..1515
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1561..1989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1648..1667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1858..1879
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1998..2018)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2003..2332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2183..2318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2245..2264
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2380..2427
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2446..2820
                     /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    2950..3071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2987..3006)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3041..3060
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3120..3136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3120..3136)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3133..3155)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3144..3160
                     /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(3168..3174,3175..3192,3193..3198))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3351..3368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3522..4110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3602..3621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4281..5072,5073..5141))
                     /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     4904..4923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5142..5246)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
       61 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
      121 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
      181 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
      241 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      301 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      361 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      421 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      481 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      541 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      601 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      661 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      721 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      781 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      841 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagccacca
      901 tggccaagat gggaaccaag gagaaggcag gaaacgcaaa gatgggcacc aaggagaaag
      961 ccgggaactg atgaattgcc aattgaaatt tgacggccta tgtcactgaa actatccaaa
     1021 caccccatgt cgatactgaa cgtatcgacg catacctcct tccttgaaaa cgcacaatca
     1081 tacaactggg cacataatgc gtacgcccat ctagtacacc catctctgta ggtccagttc
     1141 aagagctgga agagcaccct ccacttagac gtgccgtaag atagaggtta gtgagaatag
     1201 ccatccaata tgagcgttga tagctagacg aggataaggc agtgatagaa ggggcaccaa
     1261 gcttcattcg cacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1321 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1381 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1441 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     1501 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     1561 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     1621 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1681 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     1741 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     1801 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     1861 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     1921 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1981 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2041 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2101 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2161 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2221 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     2281 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     2341 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     2401 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     2461 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     2521 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     2581 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     2641 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     2701 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     2761 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     2821 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     2881 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     2941 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3001 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3061 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3121 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3181 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaacttac attaattgcg
     3241 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3301 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3361 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3421 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3481 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3541 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3601 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3661 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3721 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3781 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3841 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3901 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3961 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4021 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4081 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4141 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5221 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     5281 gg
//
