LOCUS       sequence-333624-        2721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone for assembly of multi-level gene expression controllers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 360 to 394)
  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 2721)
  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: pMLC-BB1
FEATURES             Location/Qualifiers
     source          1..2721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(360..394)
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     promoter        complement(360..394)
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     rep_origin      complement(527..1115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1226..1245)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1237..2052
                     /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             1237..2052
                     /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(1310..1329)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2211..2238)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2254..2325)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctgtcaaaa cacccccatc ttcgaacacc ataacgcgtt cccatgtaaa cccttcggga
       61 aacgactgct tgaaataatc aggcaagtca gcaggatatt tgataaagac tttgctaccg
      121 tacataaaat gtgttgccaa gatatcgaac gcaaacggta agggcccacc ttctacaact
      181 ttgatacggt tggtctgctt gccctcgtag ggtttccctt cgccttcatg cgtacatttg
      241 aattgatgac cgttcacaga accctccaaa tataacttgc tacgcatgtt ctctttgata
      301 agttcttcac ctttgctcac catctagtat ttcccctctt tctctagtac tcgagtcagg
      361 ctagcattgt acctaggact gagctagccg tcaaagttac acgggtctca tgcccaaagg
      421 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
      481 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      541 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      601 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      661 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      721 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      781 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      841 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      901 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      961 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1021 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1081 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1141 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg aacaataaaa
     1201 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     1261 tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1321 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     1381 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1441 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1501 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag
     1561 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1621 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1681 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1741 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     1801 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     1861 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     1921 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     1981 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2041 gagtttttct aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2101 acaaataggg gttccgcggt gatgagaccg taattgaata aataaaaaca gccgttgcca
     2161 gaaagaggca cggctgtttt tattttctag tgcgaccggg agcagttaaa cgcagaaagg
     2221 cccacccgaa ggtgagccag tgtgactcta gtagagagcg ttcaccgaca aacaacagat
     2281 aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggttattat
     2341 catttataca gttcatccat gcccccacca aggttgctat aacgggctac tgcgtgctcg
     2401 tattgttcga cataagtttc attgtcggcc tccttaatgc gctctaaacg gcggtcaaca
     2461 tagtgtacgc ctggcatctt tacaggcttc ttgcttttat atgtcgtctt gaagtttacg
     2521 tgtaagtgtc caccacccac aagtttcagc gccttgtcac aacgaccttc caatccaccg
     2581 tccgcaggat acatcgtctc agtcgagggc tcccatccca gggtcttttt ttgcatcacc
     2641 ggaccatttg cggggaagtt tactccgcgc acttttacgt tatagattaa ctcaccatct
     2701 tgtaagctgg tgtcctgagt c
//
