LOCUS       sequence-363783-        5480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the common wheat protein DHN_TaLEA4.
            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 5480)
  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 5480)
  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_DHN_TaLEA4 (pBS0802)
FEATURES             Location/Qualifiers
     source          1..5480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..71)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        243..622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        623..826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     777..797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     871..890
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        871..889
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1496..1513)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1502..1726
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1772..2200
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1859..1878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2069..2090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2209..2229)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2214..2543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2394..2529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2456..2475
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2591..2638
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2657..3031
                     /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    3161..3282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3198..3217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3252..3271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3331..3347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3331..3347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3344..3366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3355..3371
                     /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(3379..3385,3386..3403,3404..3409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3562..3579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3733..4321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3813..3832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4492..5283,5284..5352))
                     /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     5115..5134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5353..5457)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
       61 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      121 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
      181 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      241 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      301 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      361 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      421 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      481 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      541 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      601 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      661 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      721 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      781 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
      841 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
      901 tggctagcca ccatgtcagg ctggtttcaa gaaaagtccg gggatgctac caaaactgct
      961 tccgaaaccg gacaaacaat tcaagatcgg gcagttgaag ctaaagatca aacaggtgct
     1021 tttctgggag agaaatccga agctgttact aaagcagcaa gtgaaaccac tgaagctgct
     1081 aagaaaatgg gtggggaagc catgggaaaa gcatccgaaa ctgtcgaagc aggtaaagat
     1141 catgctgtcg aagggaaaga tcaaacgggg tctttctttg gtgataaaac tgatgctgtc
     1201 aagaagacgg caactgaaac tgcggatgct acaaaagaaa agtcaactga agctgcccaa
     1261 catgttcaag atactgctgc acaatatacg aaagatacac cagtcgcacc taaagaaaat
     1321 gtgtttcaac aagctggcgg aaatatggta ggagctgcaa caagtcgggt ccagggtagt
     1381 tgtggtggcg gtgcgactag ttttcctctc ctcagacgct ccagagtttg gccatcccga
     1441 ggagaacggc ttagtccttc ttcaagtcgg gaacccccat gaacccgctg atcagcctcg
     1501 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     1561 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     1621 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     1681 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     1741 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     1801 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1861 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1921 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1981 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2041 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2101 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2161 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     2221 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2281 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2341 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2401 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2461 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     2521 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     2581 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     2641 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
     2701 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
     2761 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
     2821 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
     2881 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
     2941 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
     3001 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
     3061 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     3121 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     3181 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3241 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     3301 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3361 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3421 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3481 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3541 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3661 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4141 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4441 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4501 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4561 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4621 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4681 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4741 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4801 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4861 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4921 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4981 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5041 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5101 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5161 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5221 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5281 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5341 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5401 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5461 atttccccga aaagtgccac
//
