LOCUS       sequence-363767-        5726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the synthetic protein 
            TJ190_negative. 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 5726)
  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 5726)
  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_TJ190_negative (pBS0773)
FEATURES             Location/Qualifiers
     source          1..5726
                     /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"
     regulatory      910..919
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1745..1762)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1751..1975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2021..2449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2108..2127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2318..2339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2458..2478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2463..2792
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2643..2778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2705..2724
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2840..2887
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2906..3280
                     /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    3410..3531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3447..3466)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3501..3520
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3580..3596)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3580..3596)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3593..3615)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3604..3620
                     /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(3628..3634,3635..3652,3653..3658))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3811..3828)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3982..4570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4062..4081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4741..5532,5533..5601))
                     /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     5364..5383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5602..5706)
                     /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 ccaccatgga tctggaggaa gtccttgacg aggctcttag ggccctggag
      961 gagttggcca gagaacgggg agacgaagag aaggtcaggg aagtggcaga agagaaaaaa
     1021 agggcggagg aggaaaaaaa agcaaataga gaagaggcga aagaactcga ggagctcata
     1081 agagagaatg agctggtgag agtgctcgtg gaggccctga gggcgcttgt cgagctcgct
     1141 gaagagcgag gcgaccttga aaaagttgaa gaagtcgcag aggaggtggc gaaggagatt
     1201 gaggagctgc gaagaaataa agaggaggct aaagagttgg ccgaactgat tagagagaat
     1261 gagcttgtta gggttttggt tgaagccctc cgcgcactcg ttgaactggc cgaggagaga
     1321 ggtgatcttg aaaaggtgga ggaggtggca gaagaagtga aagaagagat tgaagaactc
     1381 agaaggaata aggaggaagc caaagagttg gccgagctga ttagggaaaa cgagctcgta
     1441 cgcgtgcttg tggaagccct gagagccctg gtggaactcg ccgaggagag gggcgatctc
     1501 gaaaaagtgg aggaagtcgc agaggaagtt aaaagagaga ttgaggaact ccgaaggaac
     1561 aaagaggaag ctaagaagct tgctgagctc attagggaaa atgaggaggt tcgcaagaag
     1621 gtagaggagc tgcgagagtt ggtcgaaaag gcggaagaaa gaggggacct tgagaaagtt
     1681 gaggaggtcg ctaaggaagt gaaagacgaa atcgaaaggg agagggggtg aacccgctga
     1741 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1801 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1861 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1921 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     1981 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2041 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2101 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2161 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2221 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2281 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2341 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2401 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2461 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2521 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2581 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2641 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2701 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     2761 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     2821 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     2881 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     2941 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     3001 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     3061 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     3121 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     3181 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     3241 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
     3301 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3361 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3421 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3481 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3541 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3601 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3661 ctggggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     3721 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3781 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3841 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3901 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4141 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4381 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4501 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4561 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4621 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4681 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4741 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4801 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4861 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4921 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4981 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5041 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5101 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5161 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5221 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5281 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5341 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5401 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5461 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5521 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5581 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5641 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5701 tccgcgcaca tttccccgaa aagtgc
//
