LOCUS       sequence-321850-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mNeonGreen_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 6245)
  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 6245)
  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: pRD404
FEATURES             Location/Qualifiers
     source          1..6245
                     /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..2036
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1584..1593
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2279..2296)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2449..2535
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2627..2654
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2673..2764
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3222..3677
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3309..3328)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3519..3540
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4233..4892)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4807..4826
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4893..4995)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5387..5404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5521..6066)
                     /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(5601..5620)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6142..6161
                     /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 ggcgaggagg ataacatggc ctctctccca gcgacacatg
     1381 agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt cagggcaccg
     1441 gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt gacctccagt
     1501 tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac ctgccctacc
     1561 ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggctac caagtccatc
     1621 gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac acctacgagg
     1681 gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct gacggtcctg
     1741 tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaagact taccccaacg
     1801 acaaaaccat catcagtacc tttaagtgga gttacaccac tggaaatggc aagcgctacc
     1861 ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac tatctgaaga
     1921 accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc gagctcaact
     1981 tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg tacaaggctg
     2041 ccgttaaatc tggcaccaaa gctaaacgtg ctcagcgtcc ggcaaaatat agctacgttg
     2101 acgaaaacgg cgaaactaaa acctggactg gccaaggccg tactccagct gtaatcaaaa
     2161 aagcaatgga tgagcaaggt aaatccctcg acgatttcct gatcaagcaa taaagctcta
     2221 gagtcgacct gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc
     2281 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     2341 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     2401 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     2461 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2521 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     2581 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     2641 acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa
     2701 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2761 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2821 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2881 tgcagcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3001 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3181 ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct gtagcggcgc
     3241 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3301 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3361 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3421 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3481 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3541 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3601 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3661 attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3721 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3901 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4021 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     4081 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     4141 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc
     4201 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     4261 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     4321 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     4381 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     4441 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     4501 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     4561 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     4621 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc
     4681 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     4741 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     4801 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     4861 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     4921 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     4981 cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca
     5041 acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg
     5101 gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg
     5161 aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt
     5221 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     5281 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     5341 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     5401 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     5461 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     5521 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     5581 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     5641 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     5701 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     5761 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5821 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     5881 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     5941 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     6001 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     6061 tctcaagaag atcatcttat taatcagata aaatatttct aggctcatga gcccgaagtg
     6121 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     6181 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc
     6241 ttatc
//
