LOCUS       sequence-101261-        4717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control duet plasmid for split mCherry assembly assay 
            (i.e., neither half of mCherry is fused to another peptide or 
            protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4717)
  AUTHORS   Sawyer N, Gassaway BM, Haimovich AD, Isaacs FJ, Rinehart J, Regan L
  TITLE     Designed Phosphoprotein Recognition in Escherichia coli.
  JOURNAL   ACS Chem Biol. 2014 Nov 21;9(11):2502-7. doi: 10.1021/cb500658w. 
            Epub 2014 Oct 6.
  PUBMED    25272187
REFERENCE   2  (bases 1 to 4717)
  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: "10.1021/cb500658w";
            journalName: "ACS Chem Biol"; date: "2014-11-21- 21"; volume: "9"; 
            issue: "11"; pages: "2502-7"
COMMENT     Sequence Label: pNAS1B split mCherry N-link/link-C
FEATURES             Location/Qualifiers
     source          1..4717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(7..25,26..31,32..48,49..54,55..80)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    32..50
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             90..101
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(259..277)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     527..546
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      654..740
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(894..911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1028..1573)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1108..1127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1721..2599)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2626..2910
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2833..2852
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3249..3266)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3419..3505
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3597..3624
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3654..3717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3718..3786,3787..4578)
                     /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(3936..3955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agatcttccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
       61 atcagcagga cgcactgacc gaattcatta aagaggagaa aggtaccatg gcatccgtga
      121 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
      181 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      241 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
      301 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
      361 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
      421 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
      481 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
      541 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggtggctctg
      601 gctctgggtc gactggttaa tgaaagcttc atggtacgcg tgctgccagg catcaaataa
      661 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
      721 ctctcctgag taggacaaat ccgccgccct cctaggatca gcgctagcgg agtgtatact
      781 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      841 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      901 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      961 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     1021 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     1081 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     1141 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1201 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1261 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1321 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1381 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1441 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1501 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1561 aaaacgatct caagaagatc atcttattaa ctgcagagat caattcgcgc gcgaaggcga
     1621 agcggcatgc tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     1681 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     1741 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     1801 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     1861 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     1921 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     1981 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     2041 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     2101 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     2161 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     2221 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     2281 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     2341 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     2401 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     2461 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     2521 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     2581 cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc
     2641 catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac
     2701 cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa
     2761 cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg
     2821 tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct acctgacgct
     2881 ttttatcgca actctctact gtttctccat acccgttttt tgggctaaca ggaggaatga
     2941 cgtcatgggc gcggccgcag gtggaagcgg tggcgccctg aagggcgaga tcaagcagag
     3001 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     3061 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     3121 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     3181 cggcatggac gagctgtaca agtgataaga gctcttggct gttttggcgg atgagagaag
     3241 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
     3301 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
     3361 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
     3421 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3481 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     3541 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
     3601 ggccatcctg acggatggcc tttttgcgtt tctacaaact cctcgagttc aaatatgtat
     3661 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3721 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3781 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3841 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3901 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3961 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4021 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4081 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4141 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4201 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4261 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4321 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4381 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4441 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4501 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4561 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4621 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4681 aaaatccctt aacgtgagtt ttcgttccac tgagcga
//
