LOCUS       sequence-363749-        5261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the sleeping chironomid protein 
            PvLEA4_repeats_k3_11. 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 5261)
  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 5261)
  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_PvLEA4_repeats_k3_11 (pBS0761)
FEATURES             Location/Qualifiers
     source          1..5261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     866..885
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        866..884
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1272..1289)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1278..1502
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1548..1976
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1635..1654)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1845..1866
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1985..2005)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1990..2319
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2170..2305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2232..2251
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2367..2414
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2433..2807
                     /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    2937..3058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2974..2993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3028..3047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3107..3123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3107..3123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3120..3142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3131..3147
                     /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(3155..3161,3162..3179,3180..3185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3338..3355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3509..4097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3589..3608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4268..5059,5060..5128))
                     /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     4891..4910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5129..5233)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      841 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      901 agccaccatg ttccagaccg ccgccgacaa gctgggcgcc gccaaggaca ccgtggttga
      961 caccttgagg ggcgccaagg acaccgtgaa ggagaaggtg gtgggcgcca aggacgccac
     1021 cgccgactac ctgggcgaga agaaggagca gatgagggac tgatgaattg ccaattgaaa
     1081 tttgacggcc tatgtcactg aaactatcca aacaccccat gtcgatactg aacgtatcga
     1141 cgcatacctc cttccttgaa aacgcacaat catacaactg ggcacataat gcgtacgccc
     1201 atctagtaca cccatctctg taggtccagt tcaagagctg gaagagcacc ctccacttac
     1261 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1321 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1381 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1441 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1501 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1561 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1621 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1681 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1741 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1801 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1861 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1921 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1981 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2041 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2101 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2161 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2221 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2281 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2341 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     2401 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     2461 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     2521 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     2581 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     2641 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     2701 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     2761 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     2821 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2881 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     2941 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3001 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3061 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3121 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3181 gtaaagcctg gggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     3241 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3301 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3361 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3421 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3481 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3541 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3601 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3661 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3721 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3781 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3841 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3901 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3961 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4021 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4081 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4141 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4201 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4261 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4321 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4381 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4441 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4501 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4561 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4621 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4681 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4741 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4801 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4861 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4921 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4981 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5041 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5101 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5161 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5221 taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
