LOCUS       sequence-483450-        5802 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 5802)
  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 5802)
  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..5802
                     /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      1751..1798
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1836..1854)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1861..1878)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1861..1877)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1870..1892)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2119..2228
                     /note="incP origin of transfer"
     rep_origin      2418..2846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2505..2524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2715..2736
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3190..3984
                     /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(3244..3263)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3854..3873
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4002..4070,4071..4862)
                     /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(4220..4239)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5033..5621
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5522..5541
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5775..5792
                     /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 gggcgaattg gggacttcgg tcaccggcgg aatgctcagt aacacgtgga
      421 caatctgccc tcgggtgggg aataaacccg cgaaagtggg gctaataccc cataggtaca
      481 gaaggctgga atgcatctgt gcccaaagct ccggcgcccg aggatgagtc tgcggcctat
      541 caggtagtag tcagggtcac gacctgacta gccagcgacg ggtacgggcc ttgagagagg
      601 tcgcccggag atggactctg agacacgagt ccaggcccta cggggcgcag caggcgcgaa
      661 aacttcgcaa tgcaggaaac tgtgacgagg gaactccaag tgtccatgca ttgcatggac
      721 tgtttgccag cctaaaaagc tgatagagta aggggtggga aagaccggtg ccagccgccg
      781 cggtaacacc ggcacctcaa gtggtagcca ctattattgg gcctaaagcg ttcgtagccg
      841 gcttggttaa ttcctgggta aatcgtaccg ctcaacggtg cggattccgg ggacactgcc
      901 aagcttgaga ccgggggaga tcagaggtac ttgcggggga gcggtgaaat gttgtaatcc
      961 cgcagggacc accaatggcg aaggcatctg atcaaaacgg ctctgacggt gaggaacgaa
     1021 accctggggc gcaaacggga ttagataccc cggtagtcca gggcgtaaac gctgctgact
     1081 tggtgttggg ggtcctgcga gggcgtccag tgccggcacg aaggtattga gtcagccgcc
     1141 tggggagtac ggtcgcaaga ctgaaactta aaggaattgg cgggggagca ctgcaacggg
     1201 tggagcgtgc ggtttaattg gattcaacgc cggaaaactc accgggggcg acgattacat
     1261 gaaggccagg ttgatgacct tgcctgattt ttcgagaggt ggtgcatggc cgccgtcagt
     1321 tcgtaccgta aggcgttctg ttaagtcaga taacgaacga gaccctcatt gccatttgct
     1381 aggctcttct caggaagggt tgcactatgg cgataccgcc actgctgaag tggaggaagg
     1441 agggggcaac ggtaggtcag tatgccccga atctcccggg ctacacgcgc gctacaaagg
     1501 gcaggacaat gggcttcgac gccgaaaggc ggaggcaatc ccgaaacctg ctcgtacttc
     1561 ggattgaggg ctgcaattcg ccctcatgaa gctggaatcc gtagtaatcg cgggtcaaca
     1621 tcccgcggtg aatatgcccc tgctccttgc acacaccgcc cgtcaaacca ccttagttgg
     1681 gtctggacga gggctcgcct tttggcgcgc tcgaatctgg gttcagcgag gagggttaag
     1741 tcgtaacaag ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg
     1801 catgctcgag cggccgccag tgtgatggaa ttcgccctat agtgagtcgt attacaattc
     1861 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     1921 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     1981 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgctggaagg cagtacacct
     2041 tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat
     2101 ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca
     2161 ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta
     2221 tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa
     2281 atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga
     2341 agcagggtta tgcagcggaa aagcgctgct tccctgctgt tttgtggaat atctaccgac
     2401 tggaaacagg caaatgcagg aaccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2461 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2521 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2581 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2641 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2701 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2761 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2821 gatttaacaa aaatttaacg cgaattttaa caaaattcag ggcgcaaggg ctgctaaagg
     2881 aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc
     2941 tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
     3001 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
     3061 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     3121 ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatgagga
     3181 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3241 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3301 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3361 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3421 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3481 ccggggcagg atctcctgtc atcccacctt gctcctgccg agaaagtatc catcatggct
     3541 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3601 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3661 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3721 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3781 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3841 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3901 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3961 cgccttcttg acgagttctt ctgaattgaa aaaggaagag tatgagtatt caacatttcc
     4021 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4081 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4141 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4201 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4261 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4321 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4381 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4441 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4501 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4561 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4621 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4681 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4741 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4801 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4861 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4921 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4981 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5041 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5101 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5161 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5221 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5281 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5341 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5401 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5461 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5521 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5581 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5641 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5701 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5761 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//
