LOCUS       sequence-429311-        5315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptrc99A-PAmCherry-L78-PopTag for bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5315)
  AUTHORS   Hoang Y, Azaldegui CA, Dow RE, Ghalmi M, Biteen JS, Vecchiarelli AG
  TITLE     An experimental framework to assess biomolecular condensates in 
            bacteria.
  JOURNAL   Nat Commun. 2024 Apr 15;15(1):3222. doi: 10.1038/s41467-024-47330-4.
  PUBMED    38622124
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"; journalName: "Nat 
            Commun."; date: "2024-04-15"; volume: "15(1):3222. doi"; pages: " 
            10.1038/s41467-024-47330-4"
COMMENT     Sequence Label: pYH88
FEATURES             Location/Qualifiers
     source          1..5315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     primer_bind     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             267..977
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein"
                     /label=PAmCherry
                     /note="mammalian codon-optimized"
                     /translation="MAVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMK
                     FEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGAL
                     KGEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYER
                     AEGRHSTGGMDELYK"
     primer_bind     complement(415..433)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     683..702
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1499..1516)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1669..1755
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1847..1874
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1893..1984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1985..2053,2054..2845)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2203..2222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3016..3604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3505..3524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3758..3775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3790..3930
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3856..3875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4016..4038
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4116..4193
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4194..5276
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4213..4232)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgg cagtgagcaa gggcgaggag gataacatgg
      301 ccatcattaa ggagttcatg cgcttcaagg tgcacatgga ggggtccgtg aacggccacg
      361 tgttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
      421 tgaaggtgac caagggtggc cccctgccct tcacctggga catcctgtcc cctcaattca
      481 tgtacggctc caatgcctac gtgaagcacc ccgccgacat ccccgactac tttaagctgt
      541 ccttccccga gggcttcaag tgggagcgcg tgatgaaatt cgaggacggc ggcgtggtga
      601 ccgtgaccca ggactcctcc ctgcaggacg gtgagttcat ctacaaggtg aagctgcgcg
      661 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggccc
      721 tctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgaggtcaag ccgagagtga
      781 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
      841 ccgtgcagct gcccggcgcc tacaacgtca accgcaagtt ggacatcacc tcacacaacg
      901 aggactacac catcgtggaa cagtacgaac gtgccgaggg ccgccactcc accggcggca
      961 tggacgagct gtacaaggat gatgcgccgg cagagcctgc agcagaagca gccccacctc
     1021 ctcctcccga gccagaaccg gagcccgtca gcttcgacga cgaagtcttg gaattaacgg
     1081 atcccattgc tcctgaacct gagctgcccc ccttggaaac ggtgggtgat attgacgttt
     1141 acagtcctcc tgaacccgaa tctgaacccg cctatacacc gccgccagcg gctcctgtct
     1201 ttgatatggt cgcggaacaa cttgttggtg tcagtgcggc ttccgctgcc gcagcgagcg
     1261 cgttcggtag cttgagttct gctttattga tgcctaagga tggtcgcacg ttggaggacg
     1321 tggtacgtga gctgttgcgc ccgctgttaa aggagtggct tgaccagaat ctgccccgta
     1381 ttgtggagac taaagtggag gaagaggtac aacgtatctc gcgtggtcgc ggagcttaat
     1441 gagtcgacct gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc
     1501 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     1561 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     1621 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     1681 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1741 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     1801 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     1861 acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa
     1921 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1981 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2041 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2101 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2161 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2221 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2281 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2341 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2401 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2461 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2521 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2581 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2641 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2701 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2761 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2821 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2881 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2941 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3001 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3061 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3121 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3781 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3841 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     3901 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     3961 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4021 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat
     4081 tcgcgcgcga aggcgaagcg gcatgcattt acgttgacac catcgaatgg tgcaaaacct
     4141 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     4201 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     4261 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     4321 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     4381 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     4441 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     4501 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     4561 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     4621 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     4681 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     4741 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     4801 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     4861 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     4921 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     4981 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     5041 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     5101 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     5161 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5221 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5281 aacgcaatta atgtaagtta gcgcgaattg atctg
//
