LOCUS       sequence-321843-        6242 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 6242)
  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 6242)
  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..6242
                     /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      3219..3674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3306..3325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3516..3537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4230..4889)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4804..4823
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4890..4992)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5384..5401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5518..6063)
                     /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(5598..5617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6139..6158
                     /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 agttgggtgc
     2881 agcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2941 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3001 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3061 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3121 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3181 tcagaccaag tttactcata tatactttag attgatttac gcgccctgta gcggcgcatt
     3241 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3301 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3361 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3421 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3481 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3541 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     3601 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3661 aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3721 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3781 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3841 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3901 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3961 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtca
     4021 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
     4081 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct
     4141 ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt
     4201 taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac
     4261 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     4321 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     4381 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     4441 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     4501 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     4561 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     4621 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga
     4681 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     4741 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     4801 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     4861 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat
     4921 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     4981 acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca
     5041 gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg
     5101 caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt gtttttgagg
     5161 tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg acggggtggt
     5221 gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact ggcttactat
     5281 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     5341 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     5401 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     5461 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     5521 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     5581 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     5641 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     5701 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     5761 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5821 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     5881 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     5941 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     6001 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     6061 caagaagatc atcttattaa tcagataaaa tatttctagg ctcatgagcc cgaagtggcg
     6121 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     6181 gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta
     6241 tc
//
