LOCUS       sequence-192118-        4613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mNeonGreen reporter that places expression of a fluorescent protein 
            reporter (mNeonGreen) under control of the env8HyCbl riboswitch..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4613)
  AUTHORS   Polaski JT, Webster SM, Johnson JE Jr, Batey RT
  TITLE     Cobalamin Riboswitches Exhibit a Broad Range of Ability to 
            Discriminate between Methylcobalamin and Adenosylcobalamin.
  JOURNAL   J Biol Chem. 2017 May 8. pii: jbc.M117.787176. doi: 
            10.1074/jbc.M117.787176.
  PUBMED    28483920
REFERENCE   2  (bases 1 to 4613)
  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: "J Biol 
            Chem. 2017 May 8. pii: jbc.M117.787176. doi: 
            10.1074/jbc.M117.787176."
COMMENT     Sequence Label: env8 selectivity (b')
FEATURES             Location/Qualifiers
     source          1..4613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             190..900
                     /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      445..454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1240..1258)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1326..1430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1431..1499,1500..2291)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1649..1668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2462..3050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2951..2970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3204..3221
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3236..3376
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3478..3669)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4292..4378
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4470..4497
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcctgaaag cgtggtgggg aagtgacgtg aaattcgtcc agattacttg atacggttat
       61 actccgagtg ccacccaggc catacaacat acaacataca gagcaaggag actcaacatg
      121 ctgatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg
      181 gtagaaaaaa tggtgagcaa gggcgaggag gataacatgg cctctctccc agcgacacat
      241 gagttacaca tctttggctc catcaacggt gtggactttg acatggtggg tcagggcacc
      301 ggcaatccaa atgatggtta tgaggagtta aacctgaagt ccaccaaggg tgacctccag
      361 ttctccccct ggattctggt ccctcatatc gggtatggct tccatcagta cctgccctac
      421 cctgacggga tgtcgccttt ccaggccgcc atggtagatg gctccggata ccaagtccat
      481 cgcacaatgc agtttgaaga tggtgcctcc cttactgtta actaccgcta cacctacgag
      541 ggaagccaca tcaaaggaga ggcccaggtg aaggggactg gtttccctgc tgacggtcct
      601 gtgatgacca actcgctgac cgctgcggac tggtgcaggt cgaagaagac ttaccccaac
      661 gacaaaacca tcatcagtac ctttaagtgg agttacacca ctggaaatgg caagcgctac
      721 cggagcactg cgcggaccac ctacaccttt gccaagccaa tggcggctaa ctatctgaag
      781 aaccagccga tgtacgtgtt ccgtaagacg gagctcaagc actccaagac cgagctcaac
      841 ttcaaggagt ggcaaaaggc ctttaccgat gtgatgggca tggacgagct gtacaagtaa
      901 gaattccaac tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag
      961 gcctactagt cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1021 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1081 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1141 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     1201 atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg cctcgtgata cgcctatttt
     1261 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     1321 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1381 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1441 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1501 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1561 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1621 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     1681 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1741 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1801 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1861 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1921 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     1981 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2041 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2101 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2161 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2221 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2281 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2341 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2401 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2461 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2521 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2581 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2641 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2701 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2761 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2821 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2881 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2941 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3001 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3061 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3121 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3181 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3241 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     3301 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3361 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3421 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3481 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     3541 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     3601 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     3661 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     3721 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     3781 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     3841 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     3901 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     3961 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     4021 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     4081 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca
     4141 ggacccaacg ctgcccgaga atgcgcattg ttgcctggcg gcagtagcgc ggtggtccca
     4201 cctgacccca tgccgaactc agaagcgaaa cgccgtagcg ccgatggtag tgtggggtct
     4261 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     4321 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     4381 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     4441 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     4501 gctagccaca gctaacacca cgtcgtccct atctgctgcc ctaggtctat gagtggttgc
     4561 tggataactt tacgggcatg cataaggctc gtataatata ttcatataat aat
//
