LOCUS       sequence-192115-        4612 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 4612)
  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 4612)
  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 (d')
FEATURES             Location/Qualifiers
     source          1..4612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             189..899
                     /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      444..453
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1239..1257)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1325..1429
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1430..1498,1499..2290)
                     /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(1648..1667)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2461..3049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2950..2969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3203..3220
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3235..3375
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3477..3668)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4291..4377
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4469..4496
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcctgaatc atggtgggga agtgacgtga aattcgtcca gattacttga tacggttata
       61 ctccgagtgc cacccaggcc atacaacata caacatacag agcaaggaga ctcaacatgc
      121 tgatgattac gccaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccgg
      181 tagaaaaaat ggtgagcaag ggcgaggagg ataacatggc ctctctccca gcgacacatg
      241 agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt cagggcaccg
      301 gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt gacctccagt
      361 tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac ctgccctacc
      421 ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggatac caagtccatc
      481 gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac acctacgagg
      541 gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct gacggtcctg
      601 tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaagact taccccaacg
      661 acaaaaccat catcagtacc tttaagtgga gttacaccac tggaaatggc aagcgctacc
      721 ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac tatctgaaga
      781 accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc gagctcaact
      841 tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg tacaagtaag
      901 aattccaact gagcgccggt cgctaccatt accaacttgt ctggtgtcaa aaataatagg
      961 cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     1021 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     1081 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     1141 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     1201 tgcgtaagga gaaaataccg catcaggcgg ccttaagggc ctcgtgatac gcctattttt
     1261 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1321 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1381 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1441 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1501 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1561 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1621 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     1681 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1741 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1801 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1861 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1921 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     1981 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2041 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2101 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2161 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2221 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2281 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2341 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2401 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2461 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2521 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2581 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2641 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2701 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2761 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2821 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2881 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2941 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3001 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3061 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3121 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3181 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3241 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     3301 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3361 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3421 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3481 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3541 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     3601 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     3661 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     3721 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     3781 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     3841 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     3901 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     3961 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     4021 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     4081 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     4141 gacccaacgc tgcccgagaa tgcgcattgt tgcctggcgg cagtagcgcg gtggtcccac
     4201 ctgaccccat gccgaactca gaagcgaaac gccgtagcgc cgatggtagt gtggggtctc
     4261 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     4321 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     4381 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     4441 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     4501 ctagccacag ctaacaccac gtcgtcccta tctgctgccc taggtctatg agtggttgct
     4561 ggataacttt acgggcatgc ataaggctcg tataatatat tcatataata at
//
