LOCUS       sequence-321839-        6215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mEos3.2_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 6215)
  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 6215)
  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: pRD129
FEATURES             Location/Qualifiers
     source          1..6215
                     /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..2006
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     primer_bind     complement(2249..2266)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2419..2505
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2597..2624
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2643..2734
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3192..3647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3279..3298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3489..3510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4203..4862)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4777..4796
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4863..4965)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5357..5374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5491..6036)
                     /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(5571..5590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6112..6131
                     /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 gagtgcgatt aagccagaca tgaagatcaa actccgtatg gaaggcaacg
     1381 taaacgggca ccactttgtg atcgacggag atggtacagg caagcctttt gagggaaaac
     1441 agagtatgga tcttgaagtc aaagagggcg gacctctgcc ttttgccttt gatatcctga
     1501 ccactgcatt ccattacggc aacagggtat tcgccaaata tccagacaac atacaagact
     1561 attttaagca gtcgtttcct aaggggtatt cgtgggaacg aagcttgact ttcgaagacg
     1621 ggggcatttg caacgccaga aacgacataa caatggaagg ggacactttc tataataaag
     1681 ttcgatttta tggtaccaac tttcccgcca atggtccagt tatgcagaag aagacgctga
     1741 aatgggagcc ctccactgag aaaatgtatg tgcgtgatgg agtgctgacg ggtgatattg
     1801 agatggcttt gttgcttgaa ggaaatgccc attaccgatg tgacttcaga actacttaca
     1861 aagctaagga gaagggtgtc aagttaccag gcgcccactt tgtggaccac tgcattgaga
     1921 ttttaagcca tgacaaagat tacaacaagg ttaagctgta tgagcatgct gttgctcatt
     1981 ctggattgcc tgacaatgca agacgagctg ccgttaaatc tggcaccaaa gctaaacgtg
     2041 ctcagcgtcc ggcaaaatat agctacgttg acgaaaacgg cgaaactaaa acctggactg
     2101 gccaaggccg tactccagct gtaatcaaaa aagcaatgga tgagcaaggt aaatccctcg
     2161 acgatttcct gatcaagcaa taaagctcta gagtcgacct gcaggcatgc aagcttggct
     2221 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     2281 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     2341 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     2401 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2461 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     2521 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     2581 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     2641 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2701 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2761 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2821 gaaagtaaaa gatgctgaag atcagttggg tgcagcaaac tattaactgg cgaactactt
     2881 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2941 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3001 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3061 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3121 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3181 tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3241 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     3541 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3601 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa aggatctagg
     3661 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3721 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3781 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3841 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3901 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3961 catacctcgc tctgctaatc ctgttaccag tcaggcattt gagaagcaca cggtcacact
     4021 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     4081 ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta
     4141 ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa
     4201 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     4261 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     4321 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     4381 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
     4441 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     4501 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     4561 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     4621 tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg
     4681 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     4741 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     4801 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     4861 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     4921 cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc
     4981 caaaagttgg cccagggctt cccggtatca acagggacac caggatttat ttattctgcg
     5041 aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac
     5101 ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg tttctatcag
     5161 ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca
     5221 tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa
     5281 gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca
     5341 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     5401 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
     5461 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
     5521 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     5581 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     5641 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     5701 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     5761 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     5821 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     5881 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     5941 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     6001 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     6061 aaatatttct aggctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     6121 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     6181 ggcgtagagg atctgctcat gtttgacagc ttatc
//
