LOCUS       sequence-491111-        5705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Aenigmarchaeum subterraneum SCGC_AAA011-O16) 16S rRNA 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5705)
  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 5705)
  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: AenigO16
FEATURES             Location/Qualifiers
     source          1..5705
                     /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"
     terminator      complement(380..427)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1738..1757)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1739..1757)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1764..1781)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1764..1780)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1773..1795)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2022..2131
                     /note="incP origin of transfer"
     rep_origin      2321..2749
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2408..2427)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2618..2639
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3093..3887
                     /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(3147..3166)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3757..3776
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3905..3973,3974..4765)
                     /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(4123..4142)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4936..5524
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5425..5444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5678..5695
                     /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 gcatctagac catcacactg
      361 gcggccgctc gagcatgcac aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
      421 atgctagcag cgaacgatta tgacgttcgc ttactaggga ttccagcttc atgagggcga
      481 gttgcagccc tcaatctgaa ctagggctag gtttaggtga ttagctcctc ctttcggagt
      541 ggcatcacat tgtcctaacc attgtaggcc gcgtgtagcc tcgcgcaatc ggaccatacg
      601 gacctaccgt cgcccactcc ttcctctgct ttagcagcag cggtcctacc agagtgctca
      661 caatcttttg gatttctcct gggattgtga ttgcaactgg tggtgtgggt ctcgctcgtt
      721 aaccgactta acggcacgcc tcatggtacg agctgacgat ggccatgcaa tacctctcag
      781 cttgtctggt aaacccgtca ggctgacctt cattctgctg tctgcgcgag taaggtgtcc
      841 ggcgttgaat ccaattgaac cgcagcctcc accccttgtg tgctcccccg ccaattccct
      901 taagtttcag tcttgcgacc gtactcccca ggtggtccct ttaacaccta cgctccggca
      961 ctgcagtctc ccgtagagaa tgcaacacca aagggacatc gtttacagcg tggactactg
     1021 gggtatctaa tcccatttgc tccccacgct ttcgagcctc accgtcggac gcgttccgga
     1081 tggacgcctt cgccactggt ggtcctccac ggattaaagg atttaactcc tacccgtgga
     1141 atacctccat cctttcccgc tcccaagcct gatagtatct ccagcacgcc tgacagttag
     1201 ctgccagatt tcaccagaga cttgtcagac cggctacgct ctctttagac ccaataaacg
     1261 agatcaccac tcgtggcgcg ggtattaccg cggcggctgg cacccgtctt gcccaccact
     1321 tattcccctt gttttttaga caagagaaaa ggttatgcat tgcataacca ctcaagctcc
     1381 ccttctcacg cgctaacgaa ttggaaaggt ttcgcgcctg ctgcgtcccg taggacctgc
     1441 cttcatgtct cagaaggcat ctggaggctt ttactctcat aacccctacg cgttataggc
     1501 atgtgagtac tttacactcg catcaacctg atacgtcgca gtcccatcct caagctccgt
     1561 agatttcgct gtgaacgcat tccagcattt cacaactata gggtattagc ctcagtttcc
     1621 cgagattatc ccctacttga ggtcaggtta actacgtgtt actcagcagt tcggcgagcc
     1681 cagatagctc acgcctcgca tggcttagtc ggaacctgat agcagtgatc taattcgccc
     1741 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     1801 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     1861 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1921 acgcgctgga aggcagtaca ccttgatagg tgggctgccc ttcctggttg gcttggtttc
     1981 atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc ctcgcagagc
     2041 aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct
     2101 cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa
     2161 gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga
     2221 aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaagcgct gcttccctgc
     2281 tgttttgtgg aatatctacc gactggaaac aggcaaatgc aggaaccctg tagcggcgca
     2341 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc tagcgcccta
     2401 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2461 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2521 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2581 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2641 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2701 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaatt
     2761 cagggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt
     2821 gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa
     2881 agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat
     2941 ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct
     3001 gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat
     3061 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3121 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3181 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3241 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3301 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3361 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatcccac cttgctcctg
     3421 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3481 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3541 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3601 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3661 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3721 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3781 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3841 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt gaaaaaggaa
     3901 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3961 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4021 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4081 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4141 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4201 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4261 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4321 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4381 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4441 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4501 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4561 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4621 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4681 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4741 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4801 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4861 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4921 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4981 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5041 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5101 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5161 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5221 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5281 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5341 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5401 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5461 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5521 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5581 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5641 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5701 ggaag
//
