LOCUS       sequence-316026-        6236 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 6236)
  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 6236)
  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..6236
                     /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      3218..3673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3305..3324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(4228..4887)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4802..4821
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4888..4990)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5382..5399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5516..6061)
                     /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(5596..5615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6133..6152
                     /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 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2941 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3001 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3061 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3121 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3181 cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta
     3241 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3301 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3361 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3421 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3481 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aacttgaaca
     3541 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     3601 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3661 acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3721 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3781 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3841 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3901 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3961 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtcag
     4021 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     4081 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt
     4141 tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcac caggcgttta
     4201 agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg
     4261 ttgtaattca ttaagcattc tgccgacatg gaagccatca cagacggcat gatgaacctg
     4321 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     4381 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     4441 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     4501 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     4561 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     4621 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
     4681 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     4741 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     4801 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     4861 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     4921 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     4981 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
     5041 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
     5101 aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg
     5161 ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc
     5221 gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt
     5281 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
     5341 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
     5401 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
     5461 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
     5521 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     5581 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     5641 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     5701 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     5761 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5821 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     5881 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     5941 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     6001 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     6061 agaagatcat cttattaatc agataaaata tttgctcatg agcccgaagt ggcgagcccg
     6121 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     6181 gatgccggcc acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatc
//
