LOCUS       sequence-483424-        5735 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  Archael (Aenigmarchaeum subterraneum SCGC_AAA011-O16) 16S rRNA 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5735)
  TITLE     Archaeal 16S rRNA expression
REFERENCE   2  (bases 1 to 5735)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AenigO16
FEATURES             Location/Qualifiers
     source          1..5735
                     /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      1684..1731
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1769..1787)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1794..1811)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1794..1810)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1803..1825)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2052..2161
                     /note="incP origin of transfer"
     rep_origin      2351..2779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2438..2457)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2648..2669
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3123..3917
                     /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(3177..3196)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3787..3806
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3935..4003,4004..4795)
                     /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(4153..4172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4966..5554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5455..5474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5708..5725
                     /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 gggcgaatta gatcactgct atcaggttcc gactaagcca tgcgaggcgt
      421 gagctatctg ggctcgccga actgctgagt aacacgtagt taacctgacc tcaagtaggg
      481 gataatctcg ggaaactgag gctaataccc tatagttgtg aaatgctgga atgcgttcac
      541 agcgaaatct acggagcttg aggatgggac tgcgacgtat caggttgatg cgagtgtaaa
      601 gtactcacat gcctataacg cgtaggggtt atgagagtaa aagcctccag atgccttctg
      661 agacatgaag gcaggtccta cgggacgcag caggcgcgaa acctttccaa ttcgttagcg
      721 cgtgagaagg ggagcttgag tggttatgca atgcataacc ttttctcttg tctaaaaaac
      781 aaggggaata agtggtgggc aagacgggtg ccagccgccg cggtaatacc cgcgccacga
      841 gtggtgatct cgtttattgg gtctaaagag agcgtagccg gtctgacaag tctctggtga
      901 aatctggcag ctaactgtca ggcgtgctgg agatactatc aggcttggga gcgggaaagg
      961 atggaggtat tccacgggta ggagttaaat cctttaatcc gtggaggacc accagtggcg
     1021 aaggcgtcca tccggaacgc gtccgacggt gaggctcgaa agcgtgggga gcaaatggga
     1081 ttagataccc cagtagtcca cgctgtaaac gatgtccctt tggtgttgca ttctctacgg
     1141 gagactgcag tgccggagcg taggtgttaa agggaccacc tggggagtac ggtcgcaaga
     1201 ctgaaactta agggaattgg cgggggagca cacaaggggt ggaggctgcg gttcaattgg
     1261 attcaacgcc ggacacctta ctcgcgcaga cagcagaatg aaggtcagcc tgacgggttt
     1321 accagacaag ctgagaggta ttgcatggcc atcgtcagct cgtaccatga ggcgtgccgt
     1381 taagtcggtt aacgagcgag acccacacca ccagttgcaa tcacaatccc aggagaaatc
     1441 caaaagattg tgagcactct ggtaggaccg ctgctgctaa agcagaggaa ggagtgggcg
     1501 acggtaggtc cgtatggtcc gattgcgcga ggctacacgc ggcctacaat ggttaggaca
     1561 atgtgatgcc actccgaaag gaggagctaa tcacctaaac ctagccctag ttcagattga
     1621 gggctgcaac tcgccctcat gaagctggaa tccctagtaa gcgaacgtca taatcgttcg
     1681 ctgctagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcatgctc
     1741 gagcggccgc cagtgtgatg gaattcgccc tatagtgagt cgtattacaa ttcactggcc
     1801 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     1861 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1921 caacagttgc gcagcctgaa tggcgaatgg acgcgctgga aggcagtaca ccttgatagg
     1981 tgggctgccc ttcctggttg gcttggtttc atcagccatc cgcttgccct catctgttac
     2041 gccggcggta gccggccagc ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga
     2101 ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac ctatcctgcc
     2161 cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc aaaatcctgt
     2221 atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc cgaagcaggg
     2281 ttatgcagcg gaaaagcgct gcttccctgc tgttttgtgg aatatctacc gactggaaac
     2341 aggcaaatgc aggaaccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2401 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2461 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2521 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2581 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2641 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2701 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2761 caaaaattta acgcgaattt taacaaaatt cagggcgcaa gggctgctaa aggaagcgga
     2821 acacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
     2881 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
     2941 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
     3001 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     3061 caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc
     3121 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3181 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3241 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3301 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3361 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3421 aggatctcct gtcatcccac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3481 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3541 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3601 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     3661 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3721 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3781 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3841 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3901 ttgacgagtt cttctgaatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4441 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4621 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5101 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5581 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5641 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5701 cgagcgcagc gagtcagtga gcgaggaagc ggaag
//
