LOCUS       sequence-321849-        6233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mScarlet-I_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: pRD403
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..2024
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1462..1480)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1730..1749
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2267..2284)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2437..2523
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2615..2642
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2661..2752
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3210..3665
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3297..3316)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3507..3528
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4221..4880)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDIFQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4795..4814
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4881..4983)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5375..5392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5509..6054)
                     /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(5589..5608)
                     /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 agtgtgaaat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     1381 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1441 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     1501 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcatc aagcaccccg
     1561 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     1621 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     1681 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     1741 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     1801 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
     1861 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     1921 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     1981 ccgagggccg ccactccacc ggcggcatgg acgagctgta caaggctgcc gttaaatctg
     2041 gcaccaaagc taaacgtgct cagcgtccgg caaaatatag ctacgttgac gaaaacggcg
     2101 aaactaaaac ctggactggc caaggccgta ctccagctgt aatcaaaaaa gcaatggatg
     2161 agcaaggtaa atccctcgac gatttcctga tcaagcaata aagctctaga gtcgacctgc
     2221 aggcatgcaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
     2281 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
     2341 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
     2401 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
     2461 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     2521 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
     2581 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
     2641 tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct
     2701 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2761 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2821 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cagcaaacta
     2881 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3001 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3061 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3121 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3181 gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc
     3241 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3301 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3361 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3421 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3481 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3541 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3601 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     3661 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3721 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3781 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3841 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3901 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3961 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtc aggcatttga
     4021 gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata
     4081 agcggctatt taacgaccct gccctgaacc gacgaccggg tcgaatttgc tttcgaattt
     4141 ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt ttaagggcac
     4201 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     4261 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     4321 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     4381 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     4441 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     4501 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     4561 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     4621 ctatcccata tcaccagctc accgtctttc attgccatac ggaattccgg atgagcattc
     4681 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     4741 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     4801 gactgaaatg cctcaaaatg ttctttacga tgccattgga atatatcaac ggtggtatat
     4861 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     4921 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     4981 tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca
     5041 ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg
     5101 tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt
     5161 ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg
     5221 caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta tgttggcact
     5281 gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
     5341 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
     5401 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     5461 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     5521 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     5581 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     5641 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     5701 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     5761 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     5821 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     5881 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     5941 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     6001 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     6061 catcttatta atcagataaa atatttctag gctcatgagc ccgaagtggc gagcccgatc
     6121 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     6181 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atc
//
