LOCUS       sequence-483447-        5828 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 5828)
  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 5828)
  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..5828
                     /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      1777..1824
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1862..1880)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1887..1904)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1887..1903)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1896..1918)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2145..2254
                     /note="incP origin of transfer"
     rep_origin      2444..2872
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2531..2550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2741..2762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3216..4010
                     /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(3270..3289)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3880..3899
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4028..4096,4097..4888)
                     /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(4246..4265)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5059..5647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5548..5567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5801..5818
                     /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 gggcgaattc ccgactgcta tcagagtggg actaagccat gcgagtcaac
      421 gccgcaaggc gtggcatacg gctcagtaac acgtagtcaa catgcccagg ggacgtggat
      481 aacctcggga aactgaggat aaaccgcgat aggccaaggt ttctggaatg gaccatggct
      541 gaaatctata tggcccctgg attggactgc ggccgatcag cgtgttggtg aggtaatggc
      601 ccaccaaacc tgtaaccggt acgggctttg agagaaggag cccggagatg ggcactgaga
      661 caagggccca ggccctatgg ggcgcagcag gcgagaaacc tctgcaatag gcgaaacctg
      721 acagggttac tctgagtgat gcccgctaag ggctttttgg cacctctaaa aatggtgcag
      781 aataaggggt gggcaagtct ggtgtcagcc gccgcggtaa taccagcacc ccgagtggtc
      841 gggacgatta ttgggcctaa agcatccgta gcttgttctg caagtcctcc gttaaatcca
      901 cctgctcaac agatgggctg cggaggatac tacagggcta ggaggcgaga gaggcaagcg
      961 gtactcagtg ggtaggggta aaatcctctg atccaccgaa gaccaccagt ggcgaaggcg
     1021 gcttgccaga acgcgctcga cggtgaggga tgaaagctgg gggagcaaac cggattagat
     1081 acccgggtag tcccagctgt aaacgatgca gactcggtga tgggttggct tcgcgccaac
     1141 ccagtgccgc agggaagccg ttaagtctgc cgcctgggga gtacggtcgc aagactgaaa
     1201 cttaaaggaa ttggcggggg agcaccacaa ggggtgaagc ctgcggttca attggagtca
     1261 acgccgggaa tcttaccggg ggcgacagca gagtgaaggt caagctgaag actttaccag
     1321 acaagctgag aggaggtgca tggccgtcgc cagctcgtgc cgtgaggtgt cctgttaagt
     1381 caggtaacga gcaagatccc tgcctctagt tgctacctct gttctcagga acggaggggc
     1441 taattagagg gaccgccgcc gctgaggcgg aggaaggagg gggctacggc aggtcagtat
     1501 gccccgaaac cctcgggcca cacgcgggct gcaatggtaa ggacagcggg ctccgatccc
     1561 gaaaggggga ggcaatccct caaaccttac ctcagttatg attgagggct gaaactcgcc
     1621 ctcatgaata tggaatccct agtaaccgcg tgtcaccatc gcgcggtgaa tacgtccctg
     1681 ctccttgcac acaccgcccg tcgcttcacc gaagctggtt cttggcgagg tggcgcctaa
     1741 ttggcgctat cgaacctggg gtcagcaacg ggggagctag cataacccct tggggcctct
     1801 aaacgggtct tgaggggttt tttgtgcatg ctcgagcggc cgccagtgtg atggaattcg
     1861 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     1921 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     1981 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2041 tggacgcgct ggaaggcagt acaccttgat aggtgggctg cccttcctgg ttggcttggt
     2101 ttcatcagcc atccgcttgc cctcatctgt tacgccggcg gtagccggcc agcctcgcag
     2161 agcaggattc ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc
     2221 gctcgcgggt gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg
     2281 aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac
     2341 cgaaaaaatc gctataatga ccccgaagca gggttatgca gcggaaaagc gctgcttccc
     2401 tgctgttttg tggaatatct accgactgga aacaggcaaa tgcaggaacc ctgtagcggc
     2461 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2521 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2581 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2641 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2701 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2761 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2821 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2881 attcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt ccgcagaaac
     2941 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg
     3001 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt
     3061 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc
     3121 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     3181 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3241 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3301 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3361 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3421 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3481 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatcc caccttgctc
     3541 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3601 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3661 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3721 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3781 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3841 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3901 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3961 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga attgaaaaag
     4021 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4081 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4141 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4201 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4261 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4321 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4381 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4441 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4501 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4561 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4621 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4681 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4741 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4801 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4861 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4921 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4981 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5041 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5101 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5161 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5221 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5281 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5341 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5401 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5461 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5521 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5581 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5641 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5701 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     5761 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     5821 agcggaag
//
