LOCUS       sequence-342763-        6786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBS plasmid containing aadA gene with 16S rRNA promoter and psbA 3' 
            region, Nicotiana tabacum clpP1 gene with native 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 6786)
  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 6786)
  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-V4
FEATURES             Location/Qualifiers
     source          1..6786
                     /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(4601..4621)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4601..4619)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4640..4656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4640..4656)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4653..4675)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4664..4680
                     /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(4688..4694,4695..4712,4713..4718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4733..4754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4871..4888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5042..5630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5122..5141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5801..6592,6593..6661))
                     /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     6424..6443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6662..6766)
                     /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 taatgcctat tggtgttcca aaagtccctt
     3061 tccgaagtcc tggagaggaa gatgcatctt gggttgacgt atataaccga ctttatcgag
     3121 aaagattact ttttttaggc caagaggttg atagcgagat ttcgaatcaa cttattggtc
     3181 ttatggtata tctcagtatc gaggatgaga ccaaagatct gtatttgttt ataaactctc
     3241 ctgggggctg ggtaatacct ggggtggcta tttatgatac tatgcaattt gtgcgaccag
     3301 atgtccatac aatatgcatg ggattagccg cttcaatggg atcttttatc ctggtcggag
     3361 gagaaattac caaacgtcta gcattccctc acgctagggt aatgatccat caacctgcta
     3421 gttcttttta tgaggcacaa acaggcgaat ttgtcctgga agcggaagaa ctgctgaaac
     3481 tgcgtgaaac cctcacaagg gtttatgtac aaagaacggg gaaaccctta tgggttgtat
     3541 ccgaagatat ggaaagagat gtttttatgt cagcaacaga agcccaagct tatggaattg
     3601 ttgatcttgt agcggttgaa tgaaaataac atggatttcg cgcaaatctg tgatttgaga
     3661 ttttatcttc gaattgaata ttctattaaa tagaagtaat tcatcatcat tcggttagga
     3721 tcaatctaaa ccaacccatc atattatgta tacgattcaa catgccaact attaaacaac
     3781 ttattagaaa tacaagacag ccaatcagaa atgtcacgaa atcccccgct cttcggggct
     3841 gtcctcagcg tcgaggaaca tgtactaggg tgtatgtgcg actcgtttag atcatgagct
     3901 ggcacaaaag caagaaaacg acttttacag tatcaatgat cagtaccggt gaatgggata
     3961 ggatggaaga aggaactcca ttcattattt aaaaaaaact ctctcatatc cctctattgt
     4021 gtatgaagtt tatggtttcc gttggtgtaa atccaatcac agatgataag aaattctcca
     4081 ttggtaacaa atggttatcc attaagcgga ggaaatctta tttaaaaagc ataaaacata
     4141 aagattaggc aggctcccac tctggcaaga acggactaag agggtcagct acccagcaaa
     4201 ctttcataat taaataccgt tactgtatag gtagatctga ttgtgaaaga cctattactg
     4261 gataattcat gggtagagcc aaagagtgtg aactatacaa gttcccaata acatcaatta
     4321 gttgattaaa tattaaatta aagtataaag taaaggctcc ggtgtataga gaagacctca
     4381 ccgtttaaga agtaaccata gaaacgaagg aacccactat ttattttttg atttctttta
     4441 tttactagat ttttgatacc taggaaaata caatttttta tttctttctt atttcgcagt
     4501 gaaataccta aaaaaaaaaa aagaaaaaat cgtggtcggg aaggttagag tagccaaagc
     4561 cattggactc gagggggggc ccggtaccca gcttttgttc cctttagtga gggttaattg
     4621 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4681 ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga
     4741 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4801 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4861 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4921 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4981 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5041 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5101 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5161 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5221 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5281 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5341 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5401 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5461 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5521 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5581 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5641 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5701 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5761 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5821 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5881 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5941 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6001 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6061 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6121 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6181 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6241 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6301 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6361 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6421 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6481 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6541 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6601 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6661 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6721 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6781 aagtgc
//
