LOCUS       sequence-491181-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Marsarchaeota G2-cons.2) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5891)
  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 5891)
  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: MarsG2c2
FEATURES             Location/Qualifiers
     source          1..5891
                     /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(1924..1943)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1925..1943)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1950..1967)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1950..1966)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1959..1981)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2208..2317
                     /note="incP origin of transfer"
     rep_origin      2507..2935
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2594..2613)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2804..2825
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3279..4073
                     /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(3333..3352)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3943..3962
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4091..4159,4160..4951)
                     /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(4309..4328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5122..5710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5611..5630
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5864..5881
                     /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 atgctagagg aggtgatccg gccgcaggtt cccctacggc caccttgtta cgacttctcc
      481 cccctcaccg gtctcagatt cgacggaacc ggcgaaccga tcccgcctca tctgaaacca
      541 gctcgggtgg agcgacgggc ggtgtgtgca aggagcgggg acgtgttcac cgcgcgatga
      601 tgacacgcga ttactgggga ttccaggttc acgagggcgg gttgcagccc tcgatcccaa
      661 ctacggtgga gtttcaagga ttacctcccc ctttcgggtt cggaacccat tgtctccacc
      721 attgcagccc gcgtgcggcc cgggggattc ggggcatgct gacctgccgt tgccccctcc
      781 ctcctccacc ttagcggtgg cggtccccct agtgtgcccg cttgcgcggt ggcaactagg
      841 ggtgagggtc tcgctcgttc ctggacttaa ccagacacct cacggcacga gctggcgacg
      901 gccatgcgcc acctctcagc gcgtcgggca aagtcatcag cttggccctc atactgctgt
      961 cgcccccggt aaagttcccg gcgctgactc caattgagcc gcaggcttca ccccttgtgg
     1021 tgctcccccg ccaattcctt taagtttcag tcttgcgacc gtactcccca agcggcggat
     1081 ttaacggctt ccctgaggca ccgatacggc ccgaagccgt accggcaccc aatccgcatc
     1141 gtttacagct gggactaccc gggtatctaa tccggttcgc tcccccagct ttcgcccctc
     1201 accgtcggac gcgtcctagc caggcgcctt cgccactggt cgtcctcccg ggattatagg
     1261 atttcgcccc taccccggga gtacccctgg cctcttccgc tccctagccc ggcagtatcc
     1321 catgcggccc aacggttgag ccgttggatt tagcacagga cttaccggac cggctacgag
     1381 cgctttaggc tcaataatcg tcccgaccac tcgcggagct ggtattaccg cggcggctga
     1441 caccagtctt gccccccgct tattccccca ggtttttact ctggggaaaa gccagcctta
     1501 gcggctggca ctcgggatag cccagtcacg gtttcccgca ttgctaaggt ttctcgcctg
     1561 gtgcgccccg tagggcctgg gcccttgtct cagtgcccat ctccgggctc cagctctcac
     1621 tgcccgcatc cgttatcggc ttggcgagcc gttacctcgc caactacctg atggaacaca
     1681 gcctcatcct tgggcgacgc ggaagcatgc tcccacgccc tttcgatgag cacccattcc
     1741 agggttgctc gtctatggga tattagcccc agtttcccgg ggttataccc ctcccaaggg
     1801 tagattagcc gtgtgctact gagccgtacg ccacgccccc acgtggctgg aggcgttcga
     1861 ctcccatggc ttagcattat cccgatagca gtcgggtccg gcaggatcaa ccgggttaat
     1921 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     1981 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2041 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2101 gaatggacgc gctggaaggc agtacacctt gataggtggg ctgcccttcc tggttggctt
     2161 ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg
     2221 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
     2281 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
     2341 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
     2401 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa agcgctgctt
     2461 ccctgctgtt ttgtggaata tctaccgact ggaaacaggc aaatgcagga accctgtagc
     2521 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgctagc
     2581 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2641 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2701 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2761 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2821 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2881 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2941 aaaattcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga
     3001 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
     3061 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
     3121 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
     3181 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     3241 caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3301 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3361 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3421 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3481 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3541 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tcccaccttg
     3601 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3661 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3721 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3781 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3841 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3901 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3961 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4021 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaattgaaa
     4081 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4141 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4201 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4261 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4321 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4381 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4441 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4501 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4561 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4621 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4681 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4741 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4801 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4861 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4921 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4981 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5041 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5101 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5161 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5221 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5281 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5341 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5401 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5461 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5521 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5581 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5641 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5701 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5761 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5821 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5881 ggaagcggaa g
//
