LOCUS       sequence-483438-        5907 bp    DNA     circular SYN 25-AUG-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 5907)
  TITLE     Archaeal 16S rRNA expression
REFERENCE   2  (bases 1 to 5907)
  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: HyJdFR18
FEATURES             Location/Qualifiers
     source          1..5907
                     /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      1856..1903
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1941..1959)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1966..1983)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1966..1982)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1975..1997)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2224..2333
                     /note="incP origin of transfer"
     rep_origin      2523..2951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2610..2629)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2820..2841
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3295..4089
                     /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(3349..3368)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3959..3978
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4107..4175,4176..4967)
                     /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(4325..4344)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5138..5726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5627..5646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5880..5897
                     /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 ttccggttga tcctgccgga ggccactgct attggagttc
      421 gactaagcca tgcgagtcga ctggttcttc ggaaccaggg catacggctc agtaacacgt
      481 gggtaatcta ccctcgggac ggggataacc ccgggaaact ggggataata cccgataggt
      541 ggagcggtct ggaaagatgc tctgcccaaa ggctccggcg cccgaggatg agcctgcgcc
      601 cgattaggtt gttggcggtg taacggacca ccaagcctaa gatcggtacg ggcactgaga
      661 ggtggagccc ggagatgggg actgagacac ggccccaggc cctacggggc gcagcaggcg
      721 cgaaaactcc gcaatacgcg caagcgtgac gggggaactc caagtgcgcc tacgtttagt
      781 gggtgctttt gcagagtgta aacagctctg cgaataaggg ctgggtaaga ccggtgccag
      841 ccgccgcggt aacaccggca gctcaagtgg tggccgcttt tattgggcct aaagcgtccg
      901 tagccgttcc agtaagtcct tgggtaaatc ctgctcctta agagtaggaa ttctgaggat
      961 actgctggaa taggggtcga gagaggtcgg aggtactccg ggggtagcgg tgaaatgctg
     1021 taatcctcgg gggaccacca gtggcgaagg cgtccgacta gagcgagccc gacggtgagg
     1081 gacgaaagcc aggggagcga acgggattag ataccccggt agtcctggct gtaaacgctg
     1141 cgggctaggt gttgcacgct ctacgggggc gtgcagtgcc gaagagaagt tgataagccc
     1201 gccgcctggg aagtattacc gcaaggtaga aacttaaagg aattggcggg ggagcaccac
     1261 aaggggtgaa gtctgcggtt taattggatt caacgccgga aatctcactg ggggcgacgg
     1321 caggatgaag gccaagctta cgactttgct ggacgagccg agaggaggtg catggccgtc
     1381 gtcagctcgt accgtgaggc gtcctgttaa gtcaggcaac gagcgagacc cgtaccctcg
     1441 gttgtcagta gctctctgcg gagggctaga cacaacgggg ggaccgctgg cgctaagcca
     1501 gaggaaggag cgggcgacgg taggtcagta tgccccgaat cccccaggct acacgcggac
     1561 tgcaatggct aggacagagg gctccgactc cgaaaggaga aggaaatccc aaaacctagc
     1621 cccagttcgg attgtgggct ggaacccgcc cacatgaagc cggaatccct agtaatcgcg
     1681 tgtcactatc gcgcggtgaa tacgtccctg ctccttgcac acaccgcccg tcacaccacc
     1741 cgagtgaggt ccaggcaagg caatgtcctt tgggcgttgt cgagtctggg tttcgcaagg
     1801 ggggtgaagt cgtaacaagg taaccgtagc ggaagctgcg gttggatcac ctcctctagc
     1861 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcatgc tcgagcggcc
     1921 gccagtgtga tggaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt
     1981 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2041 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2101 gcgcagcctg aatggcgaat ggacgcgctg gaaggcagta caccttgata ggtgggctgc
     2161 ccttcctggt tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg
     2221 tagccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga
     2281 cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac
     2341 gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg
     2401 cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag
     2461 cggaaaagcg ctgcttccct gctgttttgt ggaatatcta ccgactggaa acaggcaaat
     2521 gcaggaaccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2581 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2641 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2701 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2761 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2821 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2881 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2941 taacgcgaat tttaacaaaa ttcagggcgc aagggctgct aaaggaagcg gaacacgtag
     3001 aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg
     3061 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga
     3121 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc
     3181 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     3241 tgatggcgca ggggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3301 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3361 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3421 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3481 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3541 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3601 ctgtcatccc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3661 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3721 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3781 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3841 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3901 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3961 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4021 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4081 ttcttctgaa ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4141 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4201 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4261 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4321 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4381 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4441 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4501 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4561 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4621 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4681 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4741 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4801 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4861 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4921 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4981 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5041 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5101 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5161 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5221 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5281 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5341 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5401 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5461 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5521 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5581 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5641 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5701 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5761 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5821 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5881 gcgagtcagt gagcgaggaa gcggaag
//
