LOCUS       sequence-366649-        5315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the rapeseed protein BnHD. 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 5315)
  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 5315)
  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_BnHD (pBS0280)
FEATURES             Location/Qualifiers
     source          1..5315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        236..615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     770..790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1284..1307
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1324..1341)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1330..1554
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1600..2028
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1687..1706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1897..1918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2037..2057)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2042..2371
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2222..2357
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2284..2303
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2419..2466
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2485..2859
                     /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    2989..3110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3026..3045)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3080..3099
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3159..3175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3159..3175)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3172..3194)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3183..3199
                     /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(3207..3213,3214..3231,3232..3237))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3390..3407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3561..4149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3641..3660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4320..5111,5112..5180))
                     /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     4943..4962
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5181..5285)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      901 ccaccatgac cgcgcaaaag gcacaacaga aagctcaaga aacagcacaa gcagctaaag
      961 ataaaacgtc tcaagcagct caaacaacac aacaaaaggc tcaagaaacg gctcaagctg
     1021 caaaagataa aacttctcaa gcagcacaaa ctacacaaca gaaagctcat gaaacaacac
     1081 aatctagcaa agaaaagact tcccaagcgg ctcaaaccgc acaagaaaag gcacgggaaa
     1141 ccaaggataa aaccggctct taccttagcg aaactgggga agctgttaaa caaaaggcac
     1201 aagatgccgc ccaatacacc aaggaaaccg cacaaaatgc agcccaatat accaaagaaa
     1261 ctgcggaagc ctaaggatct ggggactaca aggacgacga cgacaagtga acccgctgat
     1321 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1381 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1441 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     1501 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     1561 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     1621 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1681 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1741 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1801 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1861 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1921 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     1981 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     2041 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2101 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2161 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2221 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2281 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     2341 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     2401 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     2461 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     2521 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     2581 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     2641 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     2701 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     2761 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     2821 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     2881 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     2941 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     3001 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3061 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     3121 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     3181 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3241 tggggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     3301 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3361 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3421 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3481 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3541 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3601 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3661 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3721 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3781 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3841 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3901 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3961 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4021 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4081 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4141 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4201 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4261 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4321 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4381 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4441 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4501 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4561 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4621 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4681 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4741 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4801 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4861 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4921 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4981 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5041 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5101 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5161 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5221 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5281 ccgcgcacat ttccccgaaa agtgccacct gacgt
//
