LOCUS       sequence-429301-        4853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptrc99A-PAmCherry for bacterial expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4853)
  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 4853)
  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: pYH78
FEATURES             Location/Qualifiers
     source          1..4853
                     /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..980
                     /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(1037..1054)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1207..1293
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1385..1412
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1431..1522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1523..1591,1592..2383)
                     /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(1741..1760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2554..3142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3043..3062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3296..3313
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3328..3468
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3394..3413)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3554..3576
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3654..3731
                     /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             3732..4814
                     /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(3751..3770)
                     /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 gtacaagtga gtcgacctgc aggcatgcaa gcttggctgt tttggcggat
     1021 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     1081 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     1141 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     1201 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1261 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     1321 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     1381 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     1441 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1501 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1561 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1621 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1681 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1741 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     1801 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1861 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1921 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1981 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2041 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2101 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2161 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2221 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2281 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2341 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2401 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2461 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2701 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3181 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3241 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3301 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     3361 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3421 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     3481 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3541 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3601 gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcatttac gttgacacca
     3661 tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     3721 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     3781 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     3841 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     3901 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     3961 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     4021 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     4081 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     4141 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     4201 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     4261 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     4321 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     4381 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     4441 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     4501 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     4561 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     4621 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     4681 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     4741 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4801 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc gcgaattgat ctg
//
