LOCUS       sequence-366855-        5282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the sleeping chironomid protein 
            pvLEA22mer_mutant_22. 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 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_22 (pBS0718)
FEATURES             Location/Qualifiers
     source          1..5282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..49)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        221..600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        601..804
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     755..775
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     849..868
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        849..867
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      885..894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1276..1293)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1282..1506
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1552..1980
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1639..1658)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1849..1870
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1989..2009)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1994..2323
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2174..2309
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2236..2255
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2371..2418
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2437..2811
                     /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    2941..3062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2978..2997)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3032..3051
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3111..3127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3111..3127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3124..3146)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3135..3151
                     /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(3159..3165,3166..3183,3184..3189))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3342..3359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3513..4101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3593..3612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4272..5063,5064..5132))
                     /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     4895..4914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5133..5237)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
       61 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
      121 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      181 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
      241 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      301 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      361 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      421 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      481 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      541 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      601 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      661 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      721 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      781 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      841 atcgaaatta atacgactca ctatagggag acccaagctg gctagccacc atggacaagg
      901 acggaaccaa ggagaaggca ggagaggaca aggacggcac caaggagaaa gccggggagt
      961 gatgaattgc caattgaaat ttgacggcct atgtcactga aactatccaa acaccccatg
     1021 tcgatactga acgtatcgac gcatacctcc ttccttgaaa acgcacaatc atacaactgg
     1081 gcacataatg cgtacgccca tctagtacac ccatctctgt aggtccagtt caagagctgg
     1141 aagagcaccc tccacttaga cgtgccgtaa gatagaggtt agtgagaata gccatccaat
     1201 atgagcgttg atagctagac gaggataagg cagtgataga aggggcacca agcttcattc
     1261 gcacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1321 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1381 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1441 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     1501 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     1561 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1621 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1681 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1741 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     1801 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     1861 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     1921 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     1981 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2041 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2101 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2161 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2221 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2281 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     2341 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     2401 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
     2461 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     2521 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     2581 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     2641 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     2701 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     2761 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
     2821 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     2881 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     2941 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3001 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3061 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     3121 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3181 aagtgtaaag cctggggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     3241 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3301 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3361 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3421 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3481 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3541 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3601 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3661 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3721 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3781 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3841 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3901 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3961 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4021 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4081 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4141 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4201 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4261 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4321 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4381 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4441 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4501 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4561 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4621 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4681 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4741 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4801 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4861 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4921 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4981 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5041 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5101 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5161 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5221 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     5281 ga
//
