LOCUS       sequence-342769-        6939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBS plasmid containing aadA gene with 16S rRNA promoter and psbA 3' 
            region, Silene conica clpP1 gene with the tobacco regulatory 
            elements and flanking regions for tobacco plastome transformation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  AUTHORS   Abdel-Ghany SE, LaManna LM, Harroun HT, Maliga P, Sloan DB
  TITLE     Rapid sequence evolution is associated with genetic 
            incompatibilities in the plastid Clp complex.
  JOURNAL   Plant Mol Biol. 2022 Feb;108(3):277-287. doi: 
            10.1007/s11103-022-01241-4. Epub 2022 Jan 17.
  PUBMED    35039977
REFERENCE   2  (bases 1 to 6939)
  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"; doi: 
            "10.1007/s11103-022-01241-4"; journalName: "Plant Mol Biol"; date: 
            "2022-02"; volume: "108"; issue: "3"; pages: "277-287"
COMMENT     Sequence Label: pBS-V6
FEATURES             Location/Qualifiers
     source          1..6939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     680..696
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     680..696
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(1886..2674)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(4754..4774)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4754..4772)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4793..4809)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4793..4809)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4806..4828)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4817..4833
                     /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)."
     promoter        complement(join(4841..4847,4848..4865,4866..4871))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4886..4907
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5024..5041)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5195..5783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5275..5294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5954..6745,6746..6814))
                     /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     6577..6596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6815..6919)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattggagct
      661 ccaccgcggt ggcggccgct ctagaactag tggatcccag cagcgatact actggaaagg
      721 acggtttcaa tatttcccat acgcaatcct ttgtatagac gttgtggcgg acggacgcta
      781 agatggaata ggcccgctaa tatgcccaat gttcctgctg caatatgatg agaggctatt
      841 cctcctggaa caaaaggatc aaaaccttcc acgccccacg ctggatttac aggttgtact
      901 tttcccgtta gtccataagg gtcggacacc catattccgg gaccatacaa gcctgttaca
      961 tgaaatgcac caaaaccaaa gcaagccacc cctgagagaa ataaatgaat tccaaagatc
     1021 tttggcaaat ccaaagaagg ttttcctgta cgttcatcac aaaatatttc tagatcccaa
     1081 tacacccaat gccagatagc tgccaaaaag cataagccag aaaacacaat atgtgctcca
     1141 gctacacctt cgtaactcca aatacccgga ttcgttacag tcccccctgt gatactccaa
     1201 ccgccccatg aattggttat tcctaaacga gtcatgaagg gtataacgaa cataccctgt
     1261 ctccacattg gatcaagaac aggatcagaa ggatcaaaaa ccgctaattc atacagagcc
     1321 atcgaaccgg cccaaccagc aaccagagct gtatgcatta tatgaacaga aagcaaccga
     1381 ccgggatcat tcaatacaac ggtatgaaca cgataccaag gcaaacccat ggaaataccc
     1441 cttatatcaa agataaatag acactacgta actttattgc attggaaaag actataatat
     1501 gactatccta tggaccactc ttgttcagtc gattatttcc gaacaagggt gttctattct
     1561 gttggtaata atggaacaat tctgttcgta ggaacaaaga gaagcagatt tattctatac
     1621 tcgataagta ccaatacgca atgggggagt tgatcccccg ggtaaacaaa tacaaaatca
     1681 aaatagaata ctcaatcatg aataaatgca agaaaataac ctctccttct ttttctataa
     1741 tgtaaacaaa aaagtctatg taagtaaaat actagtaaat aaataaaaag aaaaaaagaa
     1801 aggagcaata gcaccctctt gatagaacaa gaaaatgatt attgctcctt tcttttcaaa
     1861 acctcctata gactaggcca ggatcttatt tgccgactac cttggtgatc tcgcctttca
     1921 cgtagtgaac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttgtccaa
     1981 gataagcctg cctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg
     2041 cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc
     2101 gggacaacgt aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata
     2161 gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt
     2221 cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag
     2281 ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc
     2341 attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca
     2401 caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt
     2461 ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa
     2521 ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat
     2581 gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt
     2641 gagtcgatac ttcggcgatc accgcttccc tcataaatcc ctccctacaa ctgtatccaa
     2701 gcgcttcgta ttcgcccgga gttcgctccc agaaatatag ccatccctgc cccctcacgt
     2761 caatcccacg agcctcttat ccattctcat tgaacgacgg cgggggagca tgccattttc
     2821 tatgagcgaa tgagtccata cttatttatc attagaaaga cctattcgta ataatttgag
     2881 tttattcatt ctgtctttct ttatgaattt ttataatcta tggataaaat aaatacgata
     2941 aaaaccaata tgaatattat aaagacaata aaaaaaattg ttacgtttcc acctcaaagt
     3001 gaaatatagt atttagttct ttctttcatt taatgcctct aggaatacca aaaggtcatt
     3061 tttcttttat agaccctgac ggagaaacga actcaacttg gacggaaata cagaactttc
     3121 tttatgaaaa aaggatactt tttattttcg aggaaattaa acccaaagtc acgtgtgaaa
     3181 ttacaggtct tctactatgt ctcagtaacg agaatgagta caaagatttt actttattta
     3241 ttaattctgc aggcggcaag acaagagatg ctctaccgat atacgacacg atgcaagtag
     3301 tggcacctgc ggtacgtaca gtaggtatcg gggtagctgc ttcaatggga gcgcttatac
     3361 tgctgggagg agaaatgggc aaacgggcag catcccctaa ttgtaggata aagattcatc
     3421 tacctgctag tgctttttat acaactgatt ctgattcgaa ttctgattct gattcgaatt
     3481 ctgattcgaa ttctgattcg aattctgata ctgataagaa aaaaaaagaa aaagacccaa
     3541 agaagaaaaa ggagcaaagt cgttcagtgg aagacttttc cgacggggta ggggaaatgt
     3601 tgcgcttgcg cgaagaattc gtaaagattt atgcaaaaag aacacaaaaa cccatatcgg
     3661 ttatagaaaa agatctagct aggggtttat ttatgtcatc agaagaagcc aaagcgtatg
     3721 gaattattga tacgctacat tacaaatttg actttaaaaa gacaattaaa aagtaaaaat
     3781 aacatggatt tcgcgcaaat ctgtgatttg agattttatc ttcgaattga atattctatt
     3841 aaatagaagt aattcatcat cattcggtta ggatcaatct aaaccaaccc atcatattat
     3901 gtatacgatt caacatgcca actattaaac aacttattag aaatacaaga cagccaatca
     3961 gaaatgtcac gaaatccccc gctcttcggg gctgtcctca gcgtcgagga acatgtacta
     4021 gggtgtatgt gcgactcgtt tagatcatga gctggcacaa aagcaagaaa acgactttta
     4081 cagtatcaat gatcagtacc ggtgaatggg ataggatgga agaaggaact ccattcatta
     4141 tttaaaaaaa actctctcat atccctctat tgtgtatgaa gtttatggtt tccgttggtg
     4201 taaatccaat cacagatgat aagaaattct ccattggtaa caaatggtta tccattaagc
     4261 ggaggaaatc ttatttaaaa agcataaaac ataaagatta ggcaggctcc cactctggca
     4321 agaacggact aagagggtca gctacccagc aaactttcat aattaaatac cgttactgta
     4381 taggtagatc tgattgtgaa agacctatta ctggataatt catgggtaga gccaaagagt
     4441 gtgaactata caagttccca ataacatcaa ttagttgatt aaatattaaa ttaaagtata
     4501 aagtaaaggc tccggtgtat agagaagacc tcaccgttta agaagtaacc atagaaacga
     4561 aggaacccac tatttatttt ttgatttctt ttatttacta gatttttgat acctaggaaa
     4621 atacaatttt ttatttcttt cttatttcgc agtgaaatac ctaaaaaaaa aaaaagaaaa
     4681 aatcgtggtc gggaaggtta gagtagccaa agccattgga ctcgaggggg ggcccggtac
     4741 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     4801 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca
     4861 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4921 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4981 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5041 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5101 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5161 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5221 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5281 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5341 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5401 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5461 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5521 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5581 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5641 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5701 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5761 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5821 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5881 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5941 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6001 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6061 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6121 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6181 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6241 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6301 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6361 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6421 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6481 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6541 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6601 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6661 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6721 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6781 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6841 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6901 aacaaatagg ggttccgcgc acatttcccc gaaaagtgc
//
