LOCUS       sequence-333681-        4349 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 4349)
  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 4349)
  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: pSTAR
FEATURES             Location/Qualifiers
     source          1..4349
                     /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      1405..1491
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1583..1610
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    2582..2601
                     /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(2607..2613,2614..2629,2630..2635))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     rep_origin      complement(2754..3342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2834..2853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3453..3472)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3464..4279
                     /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             3464..4279
                     /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(3537..3556)
                     /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 actgcttata aaacgaaagg ctcagtcgaa agactgggcc
     1441 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     1501 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     1561 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt ggcccccgac
     1621 ccttagactc tgtactcagg gaccaaaacg aaaaaaggcc cccctttcgg gaggcctctt
     1681 ttctggaatt tggtaccgag cgctagttgt atcgtacgtc ggtctaggaa tcgtttagcc
     1741 gcctcactta tacaactcat ccatgcctaa agtaatcccc gccgcggtta cgaactccaa
     1801 aaggaccatg tgatcacgtt tctcatttgg gtccttcgat aatttggatt gtgtactcag
     1861 gtagtgattg tcaggaagta aaaccgggcc gtcgccaatg ggtgtgtttt gttgataatg
     1921 gtcagccaat tgcacgctgc catcctcaat gttgtgacga atcttgaaat ttaccttaat
     1981 cccgttcttc tgcttgtcag ccatgatata cacgttgtgc gaattatagt tatactccag
     2041 tttatgtccc aaaatgttgc catcttcttt aaagtcaatg cctttcaatt caatgcgatt
     2101 cactaacgta tccccctcaa acttgacctc ggcgcgtgtc ttatagttcc cgtcatcctt
     2161 gaagaagatt gtgcgctctt gcacatagcc ttctggcatt gcagatttaa agaagtcgtg
     2221 ctgcttcatg tgatctgggt aacgcgagaa acattggacc ccgtatgtaa gtgttgtaac
     2281 cagagttggc caggggactg gaagttttcc tgtagtacaa atgaacttaa gcgtcaattt
     2341 gccgtaagta gcgtcgcctt cgccctcgcc gctaaccgaa aacttatgac cattaacgtc
     2401 cccatccaac tcaaccaaga tgggtacaac gcctgtaaac agctcctctc ctttactcac
     2461 catcagatcc ttcctcctag atccaaaaaa aaagcgggga atatatacat gaactgtata
     2521 cattccccgc tgctccaaca tttatacaac taattaaaac aattcactgt aaaaacttac
     2581 taattgtgag cgctcacaat tccacacatt atacgagccg atgattaatt gtcaacatgc
     2641 ccaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2701 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2941 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3181 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3361 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac
     3421 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     3481 ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     3541 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     3601 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3661 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3721 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc
     3781 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3841 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3901 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3961 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     4021 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     4081 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     4141 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     4201 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     4261 gctcgatgag tttttctaag aattaattca tgagcggata catatttgaa tgtatttaga
     4321 aaaataaaca aataggggtt ccgcggtga
//
