LOCUS       sequence-462335-        4046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lvl 1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4046)
  AUTHORS   Duportet X, Wroblewska L, Guye P, Li Y, Eyquem J, Rieders J, 
            Rimchala T, Batt G, Weiss R
  TITLE     A platform for rapid prototyping of synthetic gene networks in 
            mammalian cells.
  JOURNAL   Nucleic Acids Res. 2014 Dec 1;42(21):13440-51. doi: 
            10.1093/nar/gku1082. Epub 2014 Nov 5.
  PUBMED    25378321
REFERENCE   2  (bases 1 to 4046)
  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"; doi: 
            "10.1093/nar/gku1082"; journalName: "Nucleic Acids Res"; date: 
            "2014-12-1- 1"; volume: "42"; issue: "21"; pages: "13440-51"
COMMENT     Sequence Label: lvl2-[landingPad-backbone]
FEATURES             Location/Qualifiers
     source          1..4046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(19..607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(99..118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    723..760
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     protein_bind    771..795
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1407..1506)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1506..1530)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1620..1639)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1685..1740
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1739..1758)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    2149..2167
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2174..2192
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2211..2222
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2229..2906
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             complement(3097..3906)
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3820..3839
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3898..3917
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
       61 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      421 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tcctttctcg
      721 agggcttgtc gacgacggcg gtctccgtcg tcaggatcat gaattcgacc ccaagtttgt
      781 acaaaaaagc agacggcctg ccaccatgac agagtacaag ccaacagttc ggttggcaac
      841 gcgcgatgac gttccacgag ccgttaggac tcttgcagcc gcctttgcgg attaccctgc
      901 cacccggcac accgtagacc cggatcgcca catcgagcga gtgaccgaac tccaggagct
      961 tttcttgacg cgggtcgggc ttgacatagg aaaagtctgg gttgcagacg acggtgcagc
     1021 tgtggcagta tggacaacgc cggaaagcgt ggaagcaggt gctgtttttg cagaaatagg
     1081 acctcgcatg gctgagctta gtggatctcg actggcagca caacaacaaa tggaaggctt
     1141 gcttgcgcct catcgcccca aagaaccggc gtggtttctg gctacggtcg gcgtctcacc
     1201 agatcaccaa ggcaagggcc ttggttctgc cgtagtgttg cccggggtcg aggccgcaga
     1261 gagagccgga gtccctgcat tccttgagac ttctgctccg cgcaatctcc ctttctatga
     1321 aagacttgga tttaccgtta cggcggatgt tgaggtgccg gagggaccga gaacctggtg
     1381 catgacacgc aaaccagggg cctgataccc agctttcttg tacaaagttg gcattataag
     1441 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     1501 tatttaccca gctttcttgt acaaagtggt acgcgtgaat tcactcctca ggtgcaggct
     1561 gcctatcaga aggtagtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat
     1621 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     1681 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     1741 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     1801 agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt
     1861 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     1921 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     1981 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     2041 gctgtccctc ttctcttatg gagatcaccc gaagagcggt ttaccgagct cttattggtt
     2101 ttcaaacttc attgactgtg ccttaattaa tagggataac agggtaattc cctatcagtg
     2161 atagagattg acatccctat cagtgataga gatactgagc actactagag aaagaggaga
     2221 aatactagat ggcttcctcc gaggacgtta tcaaagagtt catgcgtttc aaagttcgta
     2281 tggaaggttc cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt
     2341 acgaaggtac ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt
     2401 gggacatcct gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg
     2461 acatcccgga ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga
     2521 acttcgagga cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt
     2581 tcatctacaa agttaaactg cgtggtacca acttcccgtc cgacggtccg gttatgcaga
     2641 aaaaaaccat gggttgggaa gcttccaccg aacgtatgta cccggaggac ggtgctctga
     2701 aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta
     2761 aaaccaccta catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca
     2821 aactggacat cacctcccac aacgaggact acaccatcgt tgaacagtac gaacgtgctg
     2881 aaggtcgtca ctccaccggt gcttaataat taccctgtta tccctaaccg gaatcactag
     2941 tattacagag gtaagttata acagtcgcct aaccgctctt caagagggat ggaacgacgc
     3001 gtaactcacg ttaagggatt ttggtcatga gcttgcgccg tcccgtcaag tcagcgtaat
     3061 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     3121 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     3181 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     3241 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     3301 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     3361 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     3421 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     3481 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     3541 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     3601 ttttccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     3661 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     3721 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     3781 atacaagcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     3841 atataaatca gcatccatgt tggaatttaa tcgcggcctc gacgtttccc gttgaatatg
     3901 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     3961 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacgggcc atgctagctc
     4021 cagcaaaagg ccaggaaccg taaaaa
//
