LOCUS       sequence-363992-        6011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The empty vector used for the iLID-SspB experiments in our paper.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  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 6011)
  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: empty-mCh-SspB (pBS1140)
FEATURES             Location/Qualifiers
     source          1..6011
                     /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             1671..2000
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     primer_bind     complement(2020..2037)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2026..2250
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2296..2724
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2383..2402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2593..2614
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2733..2753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2738..3067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2918..3053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2980..2999
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3115..3162
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3181..3555
                     /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    3685..3806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3722..3741)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3776..3795
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3855..3871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3855..3871)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3868..3890)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3879..3895
                     /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(3903..3909,3910..3927,3928..3933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4086..4103)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4257..4845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4337..4356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5016..5807,5808..5876))
                     /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     5639..5658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5877..5981)
                     /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 ccaccatgtc cggaggcggc ggatccggcg gaatggtgtc taaaggcgag gaggataaca
      961 tggctataat caaggaattt atgaggttca aggtgcatat ggagggatct gtgaacggtc
     1021 acgagttcga aatcgagggc gaaggcgagg ggcgacccta tgaagggaca cagacggcca
     1081 aactgaaggt gaccaagggt ggccccctgc ctttcgcctg ggacatcctg tcaccccagt
     1141 ttatgtacgg ctctaaggct tacgttaagc accctgccga tattccggac tacttgaaac
     1201 tgagcttccc tgaagggttc aagtgggaac gggtaatgaa ttttgaggat ggcggtgtag
     1261 tcacagttac ccaggacagc tctcttcagg acggagaatt tatctataag gttaaacttc
     1321 ggggcactaa cttcccatca gacggccccg tgatgcagaa gaaaactatg ggctgggaag
     1381 ccagctcaga acgcatgtat cccgaggacg gagcactgaa gggagaaatc aagcagcgac
     1441 tcaagctgaa ggatggggga cattatgatg ctgaggtcaa aaccacctat aaggccaaga
     1501 aacccgttca gcttcctggg gcatacaacg tgaatatcaa actggatatt acatcccata
     1561 acgaagacta taccatcgtg gaacagtacg agcgggccga gggcagacat agcacagggg
     1621 gcatggatga attgtacaag gcggccggat ccggtggtac ctctggaggt agctccccga
     1681 aacgccctaa gctgctgcgt gaatattacg attggctggt tgataacagc tttaccccat
     1741 atctggtggt ggatgccaca tacctgggcg tgaacgtgcc cgtggagtat gtgaaagacg
     1801 gtcagatcgt gctgaatctg tctgcaagtg cgaccggcaa cctgcaactg acaaatgatt
     1861 ttatccagtt caacgcccgc tttaagggcg tgtctcgtga actgtatatc ccgatgggtg
     1921 ccgctctggc catttacgct cgcgagaacg gcgatggtgt gatgttcgaa ccagaagaaa
     1981 tctatgacga gctgaatatt ggttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     2041 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2101 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2161 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2221 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     2281 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2341 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2401 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2461 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2521 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2581 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2641 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2701 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     2761 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2821 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2881 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2941 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3001 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3061 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     3121 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     3181 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     3241 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     3301 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     3361 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     3421 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     3481 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     3541 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3601 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3661 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3781 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     3841 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3901 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3961 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4021 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4081 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4141 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4201 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4261 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4321 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4381 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4441 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4501 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4561 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4621 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4681 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4741 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4801 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4861 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4921 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4981 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5041 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5101 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5161 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5221 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5281 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5341 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5401 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5461 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5521 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5581 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5641 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5701 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5761 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5821 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5881 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5941 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6001 ccacctgacg t
//
