LOCUS       sequence-491201-        5798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Nitrososphaera gargensis RHGA36) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5798)
  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 5798)
  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: Nitgar
FEATURES             Location/Qualifiers
     source          1..5798
                     /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(1831..1850)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1832..1850)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1857..1874)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1857..1873)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1866..1888)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2115..2224
                     /note="incP origin of transfer"
     rep_origin      2414..2842
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2501..2520)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2711..2732
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3186..3980
                     /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(3240..3259)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3850..3869
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3998..4066,4067..4858)
                     /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(4216..4235)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5029..5617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5518..5537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5771..5788
                     /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 atgctagctc ccccgttgct gaccccaggt tcgatagcgc caattaggcg ccacctcgcc
      481 aagaaccagc ttcggtgaag cgacgggcgg tgtgtgcaag gagcagggac gtattcaccg
      541 cgcgatggtg acacgcggtt actagggatt ccatattcat gagggcgagt ttcagccctc
      601 aatcataact gaggtaaggt ttgagggatt gcctccccct ttcgggatcg gagcccgctg
      661 tccttaccat tgcagcccgc gtgtggcccg agggtttcgg ggcatactga cctgccgtag
      721 ccccctcctt cctccgcctc agcggcggcg gtccctctaa ttagcccctc cgttcctgag
      781 aacagaggta gcaactagag gcagggatct tgctcgttac ctgacttaac aggacacctc
      841 acggcacgag ctggcgacgg ccatgcacct cctctcagct tgtctggtaa agtcttcagc
      901 ttgaccttca ctctgctgtc gcccccggta agattcccgg cgttgactcc aattgaaccg
      961 caggcttcac cccttgtggt gctcccccgc caattccttt aagtttcagt cttgcgaccg
     1021 tactccccag gcggcagact taacggcttc cctgcggcac tgggttggcg cgaagccaac
     1081 ccatcaccga gtctgcatcg tttacagctg ggactacccg ggtatctaat ccggtttgct
     1141 cccccagctt tcatccctca ccgtcgagcg cgttctggca agccgccttc gccactggtg
     1201 gtcttcggtg gatcagagga ttttacccct acccactgag taccgcttgc ctctctcgcc
     1261 tcctagccct gtagtatcct ccgcagccca tctgttgagc aggtggattt aacggaggac
     1321 ttgcagaaca agctacggat gctttaggcc caataatcgt cccgaccact cggggtgctg
     1381 gtattaccgc ggcggctgac accagacttg cccacccctt attctgcacc atttttagag
     1441 gtgccaaaaa gcccttagcg ggcatcactc agagtaaccc tgtcaggttt cgcctattgc
     1501 agaggtttct cgcctgctgc gccccatagg gcctgggccc ttgtctcagt gcccatctcc
     1561 gggctccttc tctcaaagcc cgtaccggtt acaggtttgg tgggccatta cctcaccaac
     1621 acgctgatcg gccgcagtcc aatccagggg ccatatagat ttcagccatg gtccattcca
     1681 gaaaccttgg cctatcgcgg tttatcctca gtttcccgag gttatccacg tcccctgggc
     1741 atgttgacta cgtgttactg agccgtatgc cacgccttgc ggcgttgact cgcatggctt
     1801 agtcccactc tgatagcagt cgggaattcg ccctatagtg agtcgtatta caattcactg
     1861 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     1921 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     1981 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgct ggaaggcagt acaccttgat
     2041 aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt
     2101 tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg
     2161 cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct
     2221 gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc
     2281 tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca
     2341 gggttatgca gcggaaaagc gctgcttccc tgctgttttg tggaatatct accgactgga
     2401 aacaggcaaa tgcaggaacc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2461 cgcagcgtga ccgctacact tgctagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2521 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2581 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     2641 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2701 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     2761 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     2821 taacaaaaat ttaacgcgaa ttttaacaaa attcagggcg caagggctgc taaaggaagc
     2881 ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     2941 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     3001 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     3061 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     3121 cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt
     3181 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3241 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3301 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3361 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3421 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3481 ggcaggatct cctgtcatcc caccttgctc ctgccgagaa agtatccatc atggctgatg
     3541 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3601 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3661 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     3721 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3781 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3841 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3901 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3961 ttcttgacga gttcttctga attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4021 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4081 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4141 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4201 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4261 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4321 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4381 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4441 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4501 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4561 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4621 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4681 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4741 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4801 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4861 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4921 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4981 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5041 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5101 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5161 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5221 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5281 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5341 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5401 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5461 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5521 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5581 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5641 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5701 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5761 gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
