LOCUS       sequence-483439-        5914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Methanoglobus hypatiae LCB24) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5914)
  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 5914)
  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: LCB24
FEATURES             Location/Qualifiers
     source          1..5914
                     /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      1863..1910
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1948..1966)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1973..1990)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1973..1989)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1982..2004)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2231..2340
                     /note="incP origin of transfer"
     rep_origin      2530..2958
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2617..2636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2827..2848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3302..4096
                     /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(3356..3375)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3966..3985
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4114..4182,4183..4974)
                     /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(4332..4351)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5145..5733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5634..5653
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5887..5904
                     /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 ttctggttga tcctgccaga ggccactgct atccggctgg
      421 gactaagcca tgcgagtcca ggggcttgcc gcaaggcaag caccggcgga cggctcagta
      481 acacgtggac aacctgccct cgggaggggg ataaccccgg gaaactgggg ctaaaccccc
      541 ataggggatg gatactggaa tgttccatct ccgaaagtcc gcaaggagcc cgaggatggg
      601 tctgcggcgg attaggtagt tggtggggta acggcccacc aagcctaaga tccgtacggg
      661 ccatgggagt gggagcccgg agatggaccc tgagacaagg gtccaggccc tacggggcgc
      721 agcaggcgcg aaacctccgc aatgcgggaa accgcgacgg ggtcagccgg agtgccgatg
      781 cttgcatcgg ctgtcgggga gcctaaaaag ctccccaagc aagggccggg caaggccggt
      841 ggcagccgcc gcggtaatac cggcggcccg agtggtggcc actattattg ggcttaaagc
      901 gtccgtagcc gggttggtta gtctcccggg aaatctggcc gcttaacggt cagaatgccg
      961 ggagatactg ccagcctagg gaccgggaga ggccgagggt attcccggag taggggtgaa
     1021 atcctataat cccgggagga ccacctgtgg cgaaggcgct cggctggaac gggtccgacg
     1081 gtgagggacg aaggccaggg gagcaaaccg gattagatac ccgggtagtc ctggctgtaa
     1141 acgatgcgga ctttgtgtcg gacaagctat gagcttgtcc ggtgccggag ggaaaccgtt
     1201 aagtccgccg cctggggagt acggccgcaa ggctgaaact taaaggaatt ggcgggggag
     1261 caccacaacg ggtggagcct gcggtttaat tagattcaac gctgggaagc ttaccggggg
     1321 agacagcggg atgaaggtcg ggctgaagac tctaccagac tagctgagag gtggtgcatg
     1381 gccgtcgtca gttcgtactg tgaagcatcc tgttaagtca ggcaacgagc gagacccgcg
     1441 ccctcagttg ccagcggttc ccttcgggga agccgggcac actgagggga ctgccggcgc
     1501 taagccggag gaaggtgcgg gcaacggcag gtccgtatgc cccgaatccc ccgggctaca
     1561 cgcgggctac aatggccggg acaatggggt ccgaccccga aaggggtagg aaatctcata
     1621 aacccggtct cacctgggat cgagggctgc aactcgccct cgtgaacctg gaatccgtag
     1681 taatcgcgtg tcaaaatcac gcggtgaata cgtccctgct ccttgcacac accgcccgtc
     1741 aagccaccca agtgggctag gggcgagggg gtgtcctttg ggcatcctcg agcccagggt
     1801 ccgcgagggg ggctaagtcg taacaaggta gccgtagggg aatctgcggc tggatcacct
     1861 ccctagcata accccttggg gcctctaaac gggtcttgag gggttttttg tgcatgctcg
     1921 agcggccgcc agtgtgatgg aattcgccct atagtgagtc gtattacaat tcactggccg
     1981 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2041 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2101 aacagttgcg cagcctgaat ggcgaatgga cgcgctggaa ggcagtacac cttgataggt
     2161 gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg
     2221 ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     2281 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     2341 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     2401 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     2461 tatgcagcgg aaaagcgctg cttccctgct gttttgtgga atatctaccg actggaaaca
     2521 ggcaaatgca ggaaccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2581 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2641 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2701 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2761 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2821 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2881 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2941 aaaaatttaa cgcgaatttt aacaaaattc agggcgcaag ggctgctaaa ggaagcggaa
     3001 cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc
     3061 tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac
     3121 atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg
     3181 ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc
     3241 aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg
     3301 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3361 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3421 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3481 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3541 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3601 ggatctcctg tcatcccacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3661 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3721 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3781 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3841 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3901 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3961 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4021 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4081 tgacgagttc ttctgaattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4141 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4201 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4261 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4321 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4381 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4441 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4501 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4561 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4621 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4681 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4741 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4801 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4861 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4921 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4981 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5041 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5101 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5161 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5221 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5281 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5341 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5401 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5461 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5521 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5581 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5641 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5701 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5761 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5821 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5881 gagcgcagcg agtcagtgag cgaggaagcg gaag
//
