LOCUS       sequence-316016-        6242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of eYFP_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: pRD396
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             join(1329..1331,1332..2042)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGINFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEGGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1371..1392)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1632..1651)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1979..2000
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2285..2302)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2455..2541
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2633..2660
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2679..2770
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3224..3679
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3311..3330)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(4234..4893)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4808..4827
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4894..4996)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5388..5405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5522..6067)
                     /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(5602..5621)
                     /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 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1381 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1441 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1501 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     1561 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1621 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1681 ccctggtgaa ccgcatcgag ctgaagggca tcaacttcaa ggaggacggc aacatcctgg
     1741 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1801 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgagggcggc agcgtgcagc
     1861 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1921 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1981 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2041 aggctgccgt taaatctggc accaaagcta aacgtgctca gcgtccggca aaatatagct
     2101 acgttgacga aaacggcgaa actaaaacct ggactggcca aggccgtact ccagctgtaa
     2161 tcaaaaaagc aatggatgag caaggtaaat ccctcgacga tttcctgatc aagcaataaa
     2221 gctctagagt cgacctgcag gcatgcaagc ttggctgttt tggcggatga gagaagattt
     2281 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
     2341 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
     2401 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
     2461 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     2521 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     2581 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     2641 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
     2701 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2761 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2821 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2881 gttggggcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2941 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3001 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3061 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3181 aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
     3241 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3301 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3361 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3421 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3481 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3541 tgaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     3601 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     3661 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3721 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3781 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3841 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3901 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3961 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4021 agtcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     4081 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat
     4141 ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg tagcaccagg
     4201 cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca
     4261 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaga cggcatgatg
     4321 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     4381 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     4441 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
     4501 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     4561 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     4621 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc
     4681 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     4741 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     4801 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
     4861 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
     4921 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     4981 tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc
     5041 aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag gtatttattc
     5101 ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt
     5161 gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg
     5221 tggtgcgtaa cggcaaaagc accgccggac atcagcgcta gcggagtgta tactggctta
     5281 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
     5341 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
     5401 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
     5461 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
     5521 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
     5581 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
     5641 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
     5701 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
     5761 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5821 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     5881 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     5941 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     6001 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     6061 atctcaagaa gatcatctta ttaatcagat aaaatatttg ctcatgagcc cgaagtggcg
     6121 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     6181 gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta
     6241 tc
//
