LOCUS       sequence-491202-        5772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Uncultured marine group III euryarchaeote clone 108exp4) 
            16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5772)
  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 5772)
  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: UMGIII
FEATURES             Location/Qualifiers
     source          1..5772
                     /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(1805..1824)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1806..1824)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1831..1848)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1831..1847)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1840..1862)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2089..2198
                     /note="incP origin of transfer"
     rep_origin      2388..2816
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2475..2494)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2685..2706
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3160..3954
                     /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(3214..3233)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3824..3843
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3972..4040,4041..4832)
                     /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(4190..4209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5003..5591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5492..5511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5745..5762
                     /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 atgctagctt gttacgactt aaccctcctc gctgaaccca gattcgagcg cgccaaaagg
      481 cgagccctcg tccagaccca actaaggtgg tttgacgggc ggtgtgtgca aggagcaggg
      541 gcatattcac cgcgggatgt tgacccgcga ttactacgga ttccagcttc atgagggcga
      601 attgcagccc tcaatccgaa gtacgagcag gtttcgggat tgcctccgcc tttcggcgtc
      661 gaagcccatt gtcctgccct ttgtagcgcg cgtgtagccc gggagattcg gggcatactg
      721 acctaccgtt gccccctcct tcctccactt cagcagtggc ggtatcgcca tagtgcaacc
      781 cttcctgaga agagcctagc aaatggcaat gagggtctcg ttcgttatct gacttaacag
      841 aacgccttac ggtacgaact gacggcggcc atgcaccacc tctcgaaaaa tcaggcaagg
      901 tcatcaacct ggccttcatg taatcgtcgc ccccggtgag ttttccggcg ttgaatccaa
      961 ttaaaccgca cgctccaccc gttgcagtgc tcccccgcca attcctttaa gtttcagtct
     1021 tgcgaccgta ctccccaggc ggctgactca ataccttcgt gccggcactg gacgccctcg
     1081 caggaccccc aacaccaagt cagcagcgtt tacgccctgg actaccgggg tatctaatcc
     1141 cgtttgcgcc ccagggtttc gttcctcacc gtcagagccg ttttgatcag atgccttcgc
     1201 cattggtggt ccctgcggga ttacaacatt tcaccgctcc cccgcaagta cctctgatct
     1261 cccccggtct caagcttggc agtgtccccg gaatccgcac cgttgagcgg tacgatttac
     1321 ccaggaatta accaagccgg ctacgaacgc tttaggccca ataatagtgg ctaccacttg
     1381 aggtgccggt gttaccgcgg cggctggcac cggtctttcc caccccttac tctatcagct
     1441 ttttaggctg gcaaacagtc catgcaatgc atggacactt ggagttccct cgtcacagtt
     1501 tcctgcattg cgaagttttc gcgcctgctg cgccccgtag ggcctggact cgtgtctcag
     1561 agtccatctc cgggcgacct ctctcaaggc ccgtacccgt cgctggctag tcaggtcgtg
     1621 accctgacta ctacctgata ggccgcagac tcatcctcgg gcgccggagc tttgggcaca
     1681 gatgcattcc agccttctgt acctatgggg tattagcccc actttcgcgg gtttattccc
     1741 cacccgaggg cagattgtcc acgtgttact gagcattccg ccggtgaccg aagtccccaa
     1801 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
     1861 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     1921 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1981 cgaatggacg cgctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct
     2041 tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc
     2101 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
     2161 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
     2221 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
     2281 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa aagcgctgct
     2341 tccctgctgt tttgtggaat atctaccgac tggaaacagg caaatgcagg aaccctgtag
     2401 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgctag
     2461 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2521 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2581 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2641 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2701 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2761 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2821 caaaattcag ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag
     2881 aaacggtgct gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca
     2941 agcgcaaaga gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg
     3001 gttttatgga cagcaagcga accggaattg ccagctgggg cgccctctgg taaggttggg
     3061 aagccctgca aagtaaactg gatggctttc ttgccgccaa ggatctgatg gcgcagggga
     3121 tcaagatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3181 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3241 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3301 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3361 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3421 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atcccacctt
     3481 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3541 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3601 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3661 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3721 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3781 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3841 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3901 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaattgaa
     3961 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4021 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4081 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4141 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4201 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4261 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4321 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4381 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4441 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4501 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4561 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4621 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4681 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4741 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4801 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4861 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4921 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4981 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5041 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5101 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5161 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5221 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5281 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5341 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5401 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5461 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5521 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5581 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5641 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5701 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5761 aggaagcgga ag
//
