LOCUS       sequence-483445-        5900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Methanomassiliicoccus luminyensis B10) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5900)
  AUTHORS   Van Beek JM, Robles G, Mewalal R, Blaby I, Hatzenpichler RO
  TITLE     A collection of archaeal 16S rRNA Clone-FISH cultures for probe 
            validation in fluorescence in situ hybridization experiments
  JOURNAL   Microbiol Resour Announc. 2025 October. 0:e01131-25.
REFERENCE   2  (bases 1 to 5900)
  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: "Microbiol 
            Resour Announc. 2025 October. 0:e01131-25."
COMMENT     Sequence Label: MlumB10
FEATURES             Location/Qualifiers
     source          1..5900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        354..372
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1849..1896
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1934..1952)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1959..1976)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1959..1975)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1968..1990)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2217..2326
                     /note="incP origin of transfer"
     rep_origin      2516..2944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2603..2622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2813..2834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3288..4082
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3342..3361)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3952..3971
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4100..4168,4169..4960)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4318..4337)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5131..5719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5620..5639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5873..5890
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg cttaagccga
      301 attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagaa ttgtaatacg
      361 actcactata gggcgaatta ttccggttga tcctgccggc ggccaccgct attggaatac
      421 gattaagaca tgcgagtcga gagacgtaag gtctcggcag accgctcagt aacacgtgga
      481 taacatgccc taaggtgggg gataatctcg ggaaactgag gataataccc cataggtcta
      541 gcatgctgga atgctactag gccgaaaact ccggtgcctt aggattggtc tgcggtctat
      601 caggcagtag tgggtgtaat ggacccacta gcccgtgacg gatatgggcc ttgagagagg
      661 gagcccagag atggactctg agacatgagt ccaggcccta cggggcgcag cagtcgcgaa
      721 aacttcgcaa tgggggaaac cccgacgagg gaattccaag tgccagcact ttgtgttggc
      781 tgttcttctg tctaaaaaac aggagaagta aggggcgggt aagacgggtg ccagccgccg
      841 cggtaatacc cgcgccccaa gtggtggtcg atattattga gcctaaaacg tccgtagccg
      901 gtcttctaaa tccttgggta aatcgggcag ctcaactgtc cgaattccgg ggagactggg
      961 agacttggga ccgggagagg tcagaggtac ttctggggta ggggtaaaat cctgtaatcc
     1021 tggaaggacc accggtggcg aaggcgtctg actagaacgg atccgacggt gagggacgaa
     1081 gccctggggc gcaaacggga ttagataccc cggtagtcca gggtgtaaac gctgcgggct
     1141 tggtgttggg ggtccttagt gggcgcccag tgccggagag aagttgttaa gcctgctgct
     1201 tggggagtat gtccgcaagg atgaaactta aaggaattgg cgggggagca ccgcaacggg
     1261 aggagcgtgc ggttcaattg gattcaacgc cgaaaaactc accaggagcg acggatatat
     1321 gaaggtcaag ccgatgactt tacctgattt tccgagaggt ggtgcatggc cgtcgtcagt
     1381 tcgtaccgta aggcgttctc ttaagtgaga taacgaacga gaccctcacc attaattgct
     1441 agtcccttct ccggaggggg cgcacattaa tgggaccgct ggcgctaagt cagaggaagg
     1501 agaggtcaac ggtaggtccg tatgccccga atcccctggg caacacgcgc gctacaaagg
     1561 acgggacaat gggcttcgac accgaaaggt ggagacaatc ctgaaacccg tccatagttc
     1621 ggattggggg ctgtaacccg cccccatgaa gctggattcc gtagtaatcg cttgtcaaca
     1681 tcaagcggtg aatatgcccc tgctccttgc acacaccgcc cgtcaaacca cccgagttgg
     1741 gtttcagtga ggatgtttct tattggggct ttcgaactga gattcagcaa ggagggttaa
     1801 gtcgtaacaa ggtatctgta ggggaacctg cagatggatc acctcctact agcataaccc
     1861 cttggggcct ctaaacgggt cttgaggggt tttttgtgca tgctcgagcg gccgccagtg
     1921 tgatggaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     1981 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     2041 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     2101 ctgaatggcg aatggacgcg ctggaaggca gtacaccttg ataggtgggc tgcccttcct
     2161 ggttggcttg gtttcatcag ccatccgctt gccctcatct gttacgccgg cggtagccgg
     2221 ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa
     2281 gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg
     2341 gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa
     2401 ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaaa
     2461 gcgctgcttc cctgctgttt tgtggaatat ctaccgactg gaaacaggca aatgcaggaa
     2521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2881 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2941 aattttaaca aaattcaggg cgcaagggct gctaaaggaa gcggaacacg tagaaagcca
     3001 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     3061 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     3121 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta
     3181 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc
     3241 gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3301 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3361 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3421 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3481 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3541 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3601 cccaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3661 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3721 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3781 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     3841 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3901 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3961 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4021 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4081 gaattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4141 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4201 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4261 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4321 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4381 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4441 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4501 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4561 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4621 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4681 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4741 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4801 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4861 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4921 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4981 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5041 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5101 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5161 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5221 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5281 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5341 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5401 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5461 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5521 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5581 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5641 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5701 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5761 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5821 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5881 agtgagcgag gaagcggaag
//
