LOCUS       sequence-491177-        5877 bp    DNA     circular SYN 30-OCT-2025
DEFINITION  Archael (Hydrothermarchaeota archaeon JdFR-18) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5877)
  TITLE     Archaeal 16S rRNA expression
REFERENCE   2  (bases 1 to 5877)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: HyJdFR18
FEATURES             Location/Qualifiers
     source          1..5877
                     /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(1910..1929)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1911..1929)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1936..1953)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1936..1952)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1945..1967)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2194..2303
                     /note="incP origin of transfer"
     rep_origin      2493..2921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2580..2599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2790..2811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3265..4059
                     /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(3319..3338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3929..3948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4077..4145,4146..4937)
                     /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(4295..4314)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5108..5696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5597..5616
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5850..5867
                     /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 aggtgatcca accgcagctt ccgctacggt taccttgtta cgacttcacc
      481 ccccttgcga aacccagact cgacaacgcc caaaggacat tgccttgcct ggacctcact
      541 cgggtggtgt gacgggcggt gtgtgcaagg agcagggacg tattcaccgc gcgatagtga
      601 cacgcgatta ctagggattc cggcttcatg tgggcgggtt ccagcccaca atccgaactg
      661 gggctaggtt ttgggatttc cttctccttt cggagtcgga gccctctgtc ctagccattg
      721 cagtccgcgt gtagcctggg ggattcgggg catactgacc taccgtcgcc cgctccttcc
      781 tctggcttag cgccagcggt ccccccgttg tgtctagccc tccgcagaga gctactgaca
      841 accgagggta cgggtctcgc tcgttgcctg acttaacagg acgcctcacg gtacgagctg
      901 acgacggcca tgcacctcct ctcggctcgt ccagcaaagt cgtaagcttg gccttcatcc
      961 tgccgtcgcc cccagtgaga tttccggcgt tgaatccaat taaaccgcag acttcacccc
     1021 ttgtggtgct cccccgccaa ttcctttaag tttctacctt gcggtaatac ttcccaggcg
     1081 gcgggcttat caacttctct tcggcactgc acgcccccgt agagcgtgca acacctagcc
     1141 cgcagcgttt acagccagga ctaccggggt atctaatccc gttcgctccc ctggctttcg
     1201 tccctcaccg tcgggctcgc tctagtcgga cgccttcgcc actggtggtc ccccgaggat
     1261 tacagcattt caccgctacc cccggagtac ctccgacctc tctcgacccc tattccagca
     1321 gtatcctcag aattcctact cttaaggagc aggatttacc caaggactta ctggaacggc
     1381 tacggacgct ttaggcccaa taaaagcggc caccacttga gctgccggtg ttaccgcggc
     1441 ggctggcacc ggtcttaccc agcccttatt cgcagagctg tttacactct gcaaaagcac
     1501 ccactaaacg taggcgcact tggagttccc ccgtcacgct tgcgcgtatt gcggagtttt
     1561 cgcgcctgct gcgccccgta gggcctgggg ccgtgtctca gtccccatct ccgggctcca
     1621 cctctcagtg cccgtaccga tcttaggctt ggtggtccgt tacaccgcca acaacctaat
     1681 cgggcgcagg ctcatcctcg ggcgccggag cctttgggca gagcatcttt ccagaccgct
     1741 ccacctatcg ggtattatcc ccagtttccc ggggttatcc ccgtcccgag ggtagattac
     1801 ccacgtgtta ctgagccgta tgccctggtt ccgaagaacc agtcgactcg catggcttag
     1861 tcgaactcca atagcagtgg cctccggcag gatcaaccgg aataattcgc cctatagtga
     1921 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     1981 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2041 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgctg
     2101 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     2161 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     2221 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     2281 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     2341 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     2401 ctataatgac cccgaagcag ggttatgcag cggaaaagcg ctgcttccct gctgttttgt
     2461 ggaatatcta ccgactggaa acaggcaaat gcaggaaccc tgtagcggcg cattaagcgc
     2521 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gctagcgccc tagcgcccgc
     2581 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2641 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2701 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2761 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2821 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2881 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa ttcagggcgc
     2941 aagggctgct aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc
     3001 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
     3061 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
     3121 agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta
     3181 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctgatcaa
     3241 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3301 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3361 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3421 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3481 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3541 ctattgggcg aagtgccggg gcaggatctc ctgtcatccc accttgctcc tgccgagaaa
     3601 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3661 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3721 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3781 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3841 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3901 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3961 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4021 cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttgaaaaagg aagagtatga
     4081 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4141 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4201 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4261 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4321 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4381 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4441 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4501 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4561 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4621 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4681 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4741 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4801 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4861 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4921 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4981 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5041 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5101 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5161 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5221 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5281 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5341 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5401 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5461 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5521 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5581 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5641 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5701 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5761 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5821 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
