LOCUS       sequence-316018-        6233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mCherry_H-NSdbd in Escherichia coli (and, potentially,
            other bacteria).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6233)
  AUTHORS   Rashid FM, Mahlandt E, van der Vaart M, Boer DEC, Varela Alvarez M, 
            Henneman B, Brocken DJW, Voskamp P, Blok AJ, Shimizu TS, Meijer AH, 
            Luijsterburg MS, Goedhart J, Cremazy FGE, Dame RT
  TITLE     HI-NESS: a family of genetically encoded DNA labels based on a 
            bacterial nucleoid-associated protein.
  JOURNAL   Nucleic Acids Res. 2021 Nov 4. pii: 6414584. doi: 
            10.1093/nar/gkab993.
  PUBMED    34734265
REFERENCE   2  (bases 1 to 6233)
  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: "Nucleic 
            Acids Res. 2021 Nov 4. pii: 6414584. doi: 10.1093/nar/gkab993."
COMMENT     Sequence Label: pRD395
FEATURES             Location/Qualifiers
     source          1..6233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1329..2033
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1471..1489)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1739..1758
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2276..2293)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2446..2532
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2624..2651
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2670..2761
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3215..3670
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3302..3321)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(4225..4884)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4799..4818
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4885..4987)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5379..5396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5513..6058)
                     /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(5593..5612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6130..6149
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcgaatt cgagctcagg
     1321 agtgtgaagt gagcaagggc gaggaggata acatggctat catcaaggag ttcatgcgct
     1381 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     1441 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     1501 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     1561 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     1621 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     1681 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     1741 ccgtaatgca gaagaagact atgggctggg aggcctcctc cgagcggatg taccccgagg
     1801 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     1861 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1921 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1981 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aaggctgccg
     2041 ttaaatctgg caccaaagct aaacgtgctc agcgtccggc aaaatatagc tacgttgacg
     2101 aaaacggcga aactaaaacc tggactggcc aaggccgtac tccagctgta atcaaaaaag
     2161 caatggatga gcaaggtaaa tccctcgacg atttcctgat caagcaataa agctctagag
     2221 tcgacctgca ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     2281 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     2341 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     2401 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     2461 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     2521 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     2581 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
     2641 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat
     2701 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2761 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2821 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttggggca
     2881 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2941 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3001 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3061 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3121 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3181 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc
     3241 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3301 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3361 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3421 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3481 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac ttgaacaaca
     3541 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3601 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3661 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3721 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3781 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3841 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3901 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3961 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtcaggca
     4021 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     4081 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg
     4141 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag gcgtttaagg
     4201 gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg
     4261 taattcatta agcattctgc cgacatggaa gccatcacag acggcatgat gaacctgaat
     4321 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     4381 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     4441 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     4501 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     4561 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     4621 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc
     4681 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     4741 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     4801 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     4861 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     4921 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     4981 ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac
     5041 accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag
     5101 tgcgtcgggt gatgctgcca acttactgat ttagtgtatg atggtgtttt tgaggtgctc
     5161 cagtggcttc tgtttctatc agctgtccct cctgttcagc tactgacggg gtggtgcgta
     5221 acggcaaaag caccgccgga catcagcgct agcggagtgt atactggctt actatgttgg
     5281 cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg
     5341 cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga ctcgctacgc
     5401 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     5461 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     5521 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     5581 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
     5641 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     5701 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     5761 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     5821 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     5881 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     5941 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
     6001 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     6061 agatcatctt attaatcaga taaaatattt gctcatgagc ccgaagtggc gagcccgatc
     6121 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     6181 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atc
//
