LOCUS       sequence-483449-        5470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (13cm_4210_3138-4177(-) _Thor) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5470)
  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 5470)
  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: Thor-LSSM13-A
FEATURES             Location/Qualifiers
     source          1..5470
                     /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      1419..1466
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1504..1522)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1529..1546)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1529..1545)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1538..1560)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            1787..1896
                     /note="incP origin of transfer"
     rep_origin      2086..2514
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2173..2192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2383..2404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2858..3652
                     /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(2912..2931)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3522..3541
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3670..3738,3739..4530)
                     /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(3888..3907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4701..5289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5190..5209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5443..5460
                     /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 tccgtaaggt ggaccgctag aaaggagagg gcaaggctgg
      421 tgccagccgc cgcggtaaaa ccagctcttc gagtggtgtc catgattatt gagcttaaag
      481 cgttcgtagc cggttctgta ggtccctgct taaatctggc cgctcaacgg tcagtctggt
      541 agggataccg ctttacttgg ggacgggaga cggcgacggt attccgtagg taagggtgaa
      601 atctaatgat ctacggagga ccaccagtgg cgaaggcggt cgtcgagaac gtgcccgacg
      661 gtgagggacg aaacccaggg gagcaaatcg gattagatac ccgagtagtc ctgggcgtaa
      721 acgatgtccg ttaagtgttg cattggccac gagccactgc ggtgctgtag cgaaagcgat
      781 aaacggaccg cttgggaagt acggtcgcaa gactgaatta gagagccatt ccgactctcc
      841 tcaggagact taaaggaatt ggcgggggag caccacaagg ggtggagctt gcggtttaat
      901 tggactcaac gccggaaatc ttacccgccc agacagccga atgagagcca gactgatgat
      961 cttgctcgac gagctgagag ctggtgcatg gccatcgtaa attcgtgccg tgaggtatca
     1021 gcttaagtgc tgcaacgaat gagatccctg ccacctagtt gccagccggg ggcttcggcc
     1081 ctgacggggc acactagggg gaccgccagc gaaaagctgg aggaaggtgg gggccacggt
     1141 aggtcagtat gccgcgatat gggcggggcc acacgcgtgc tacaatggct gggacaatgc
     1201 gtcacaacac cgaaaggtga agtcaatcgc gaaacccagt cgtagtcggg atcgcgggtt
     1261 ggaactcacc cgcgtgaacc tggatttcac attcacttcg aatgtggaca ggagatccct
     1321 agtatccgcg tctcactatg gcgcggagaa tacgtccctg cttcttgcac acaccgcccg
     1381 tcgctccatg cgagcggcct tggggtgagg tgtcgtccct agcataaccc cttggggcct
     1441 ctaaacgggt cttgaggggt tttttgtgca tgctcgagcg gccgccagtg tgatggaatt
     1501 cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg
     1561 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     1621 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     1681 aatggacgcg ctggaaggca gtacaccttg ataggtgggc tgcccttcct ggttggcttg
     1741 gtttcatcag ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc
     1801 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac
     1861 ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa
     1921 ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat
     1981 accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaaa gcgctgcttc
     2041 cctgctgttt tgtggaatat ctaccgactg gaaacaggca aatgcaggaa ccctgtagcg
     2101 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2161 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2221 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2281 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2341 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2401 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2461 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2521 aaattcaggg cgcaagggct gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa
     2581 acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
     2641 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
     2701 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
     2761 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
     2821 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     2881 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     2941 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3001 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3061 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3121 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat cccaccttgc
     3181 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3241 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3301 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3361 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3421 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3481 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3541 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3601 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaattgaaaa
     3661 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3721 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3781 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3841 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3901 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3961 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4021 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4081 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4141 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4201 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4261 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4321 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4381 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4441 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4501 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4561 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4621 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4681 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4741 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4801 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4861 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4921 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4981 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5041 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5101 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5161 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5221 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5281 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5341 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5401 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5461 gaagcggaag
//
