LOCUS       sequence-366759-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the nematode (Aphelenchus avenae) 
            protein LEA1_APHAV. 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 5378)
  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 5378)
  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_LEA1_APHAV (pBS0335)
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..76)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        248..627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        628..831
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     782..802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     876..895
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        876..894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1359..1382
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1399..1416)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1405..1629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1675..2103
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1762..1781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1972..1993
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2112..2132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2117..2446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2297..2432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2359..2378
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2494..2541
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2560..2934
                     /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    3064..3185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3101..3120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3155..3174
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3234..3250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3234..3250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3247..3269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3258..3274
                     /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(3282..3288,3289..3306,3307..3312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3465..3482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3636..4224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3716..3735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4395..5186,5187..5255))
                     /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     5018..5037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5256..5360)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
       61 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
      121 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
      181 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      241 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      301 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      361 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      421 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      481 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      541 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      601 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      661 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      721 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      781 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      841 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      901 caagctggct agccaccatg tcatcacaac agaatcagaa cagacagggc gagcagcagg
      961 agcagggcta catggaggcc gctaaggaga aggtggtgaa cgcctgggag tccaccaagg
     1021 agaccctgag cagcactgcc caggccgccg ccgagaagac cgctgaattc agagacagcg
     1081 ccggcgaaac catcagggac ctcaccggcc aggcccagga gaaggggcag gagttcaagg
     1141 agagagccgg cgagaaggct gaggagacca agcagagggc cggcgagaaa atggacgaga
     1201 ccaagcagag agccggtgag atgcgcgaga acgccgggca gaagatggag gagtataagc
     1261 agcagggcaa gggcaaggcc gaggagctgc gggacaccgc tgccgagaag ctgcaccagg
     1321 ccggcgagaa ggtgaagggc agagactgag gatctgggga ctacaaggac gacgacgaca
     1381 agtgaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1441 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1501 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     1561 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     1621 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     1681 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     1741 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1801 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1861 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1921 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1981 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2041 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2101 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2161 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2221 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2281 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2341 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     2401 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     2461 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     2521 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
     2581 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
     2641 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
     2701 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
     2761 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
     2821 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
     2881 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
     2941 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3001 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3061 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3121 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3181 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3241 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3301 ataaagtgta aagcctgggg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     3361 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3421 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3481 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3541 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3601 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3661 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3721 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3781 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3841 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3901 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3961 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4021 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4081 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4141 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4201 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4261 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4321 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4381 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4441 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4501 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4561 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4621 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4681 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4741 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4801 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4861 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4921 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4981 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5041 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5101 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5161 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5221 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5281 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5341 aaacaaatag gggttccgcg cacatttccc cgaaaagt
//
