LOCUS       sequence-363763-        5261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the sleeping chironomid protein 
            PvLEA4_repeats_k5_15. This is one of the bottom performing targets 
            in our apoptosis assay (it induced 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_k5_15 (pBS0767)
FEATURES             Location/Qualifiers
     source          1..5261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..74)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        626..829
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     874..893
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        874..892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1280..1297)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1286..1510
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1556..1984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1643..1662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1853..1874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1993..2013)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1998..2327
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2178..2313
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2240..2259
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2375..2422
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2441..2815
                     /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    2945..3066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2982..3001)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3036..3055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3115..3131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3115..3131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3128..3150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3139..3155
                     /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(3163..3169,3170..3187,3188..3193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3346..3363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3517..4105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3597..3616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4276..5067,5068..5136))
                     /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     4899..4918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5137..5241)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
       61 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      121 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      181 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      241 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      841 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      901 agctggctag ccaccatgtt ccagaccgcc gccgacaagc tgggcgccgc caaggacacc
      961 gtggttgaca ccttggtgga cgccaaggac accgtgaagg agaaggccgt gggcgccaag
     1021 gacgccaccg ccgactacct tggagaggct aaggagcaga tgagggactg atgaattgcc
     1081 aattgaaatt tgacggccta tgtcactgaa actatccaaa caccccatgt cgatactgaa
     1141 cgtatcgacg catacctcct tccttgaaaa cgcacaatca tacaactggg cacataatgc
     1201 gtacgcccat ctagtacacc catctctgta ggtccagttc aagagctgga agagcaccct
     1261 ccacttaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1321 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1381 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1441 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1501 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     1561 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1621 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1681 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1741 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1801 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1861 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1921 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1981 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2041 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2101 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2161 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2221 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2281 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     2341 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     2401 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
     2461 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
     2521 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
     2581 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
     2641 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
     2701 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
     2761 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
     2821 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     2881 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     2941 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3001 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3061 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3121 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3181 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     3241 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3301 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3361 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3421 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3481 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3541 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3601 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3661 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3721 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3781 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3841 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3901 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3961 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4021 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4081 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4141 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4201 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4261 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4321 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4381 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4441 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4501 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4561 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4621 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4681 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4741 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4801 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4861 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4921 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4981 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5041 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5101 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5161 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5221 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg c
//
