LOCUS       sequence-333684-        4369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  STAR-based multi-level gene expression controller (MLC).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4369)
  AUTHORS   Greco FV, Pandi A, Erb TJ, Grierson CS, Gorochowski TE
  TITLE     Harnessing the central dogma for stringent multi-level control of 
            gene expression.
  JOURNAL   Nat Commun. 2021 Mar 19;12(1):1738. doi: 10.1038/s41467-021-21995-7.
  PUBMED    33741937
REFERENCE   2  (bases 1 to 4369)
  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: "2021-03-19"; volume: "12(1):1738. doi"; pages: " 
            10.1038/s41467-021-21995-7"
COMMENT     Sequence Label: pSTAR2
FEATURES             Location/Qualifiers
     source          1..4369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(57..1139)
                     /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="MKPVTLYDVAEYAGVSYMTVSRVVNQASHVSAKTREKVEAAMVEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIYSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1101..1120
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1278..1283,1284..1299,1300..1306)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1312..1331
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     terminator      1415..1501
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1593..1620
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    2602..2621
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        complement(join(2627..2633,2634..2649,2650..2655))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     rep_origin      complement(2774..3362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2854..2873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3473..3492)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3484..4299
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3484..4299
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3557..3576)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 agtgaaaaga aaaaaggccg cagagcggcc tttttagtta gatcggaccc cacatctcac
       61 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
      121 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagact
      181 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
      241 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
      301 gagctatctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
      361 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
      421 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
      481 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
      541 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
      601 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcctc atgggagtaa
      661 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
      721 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
      781 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
      841 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
      901 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
      961 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc caccatcgcc
     1021 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     1081 acggtcatat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcata
     1141 ttcacctcca tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttt
     1201 caccattcga tggcgcgccg cggctgggaa tctgagacat gagtcaagat atttgctcgg
     1261 taacgtatgc tggagtgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
     1321 cgctcacaat taggttgaac tgtatacatt ccccgctgct ccaacattta tacaactaat
     1381 taaaacaatt cactgtaaaa acttttccta gacgcttata aaacgaaagg ctcagtcgaa
     1441 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     1501 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     1561 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
     1621 ggcccccgac ccttagactc tgtactcagg gaccaaaacg aaaaaaggcc cccctttcgg
     1681 gaggcctctt ttctggaatt tggtaccgag cgctagttgt atcgtacgtc ggtctaggaa
     1741 tcgtttagcc gcctcactta tacaactcat ccatgcctaa agtaatcccc gccgcggtta
     1801 cgaactccaa aaggaccatg tgatcacgtt tctcatttgg gtccttcgat aatttggatt
     1861 gtgtactcag gtagtgattg tcaggaagta aaaccgggcc gtcgccaatg ggtgtgtttt
     1921 gttgataatg gtcagccaat tgcacgctgc catcctcaat gttgtgacga atcttgaaat
     1981 ttaccttaat cccgttcttc tgcttgtcag ccatgatata cacgttgtgc gaattatagt
     2041 tatactccag tttatgtccc aaaatgttgc catcttcttt aaagtcaatg cctttcaatt
     2101 caatgcgatt cactaacgta tccccctcaa acttgacctc ggcgcgtgtc ttatagttcc
     2161 cgtcatcctt gaagaagatt gtgcgctctt gcacatagcc ttctggcatt gcagatttaa
     2221 agaagtcgtg ctgcttcatg tgatctgggt aacgcgagaa acattggacc ccgtatgtaa
     2281 gtgttgtaac cagagttggc caggggactg gaagttttcc tgtagtacaa atgaacttaa
     2341 gcgtcaattt gccgtaagta gcgtcgcctt cgccctcgcc gctaaccgaa aacttatgac
     2401 cattaacgtc cccatccaac tcaaccaaga tgggtacaac gcctgtaaac agctcctctc
     2461 ctttactcac catcagatcc ttcctcctag atccaaaaaa aaagcgggga atatatacat
     2521 gaactgtata cattccccgc tgctccaaca tttatacaac taattaaaac aattcactgt
     2581 aaaaactttt cctagactac taattgtgag cgctcacaat tccacacatt atacgagccg
     2641 atgattaatt gtcaacatgc ccaaaggcgg taatacggtt atccacagaa tcaggggata
     2701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2941 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3181 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3421 aagggatttt ggtcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     3481 gttatgagcc atattcaacg ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg
     3541 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     3601 atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     3661 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     3721 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     3781 gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     3841 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     3901 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     3961 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     4021 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     4081 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     4141 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     4201 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     4261 aaattgcagt ttcatttgat gctcgatgag tttttctaag aattaattca tgagcggata
     4321 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcggtga
//
