LOCUS       sequence-405336-        8297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  amyE::(Ppen-lacI-Delta-11-dronPA cm). PALM integration vector 
            includes dronPA fusion tagged C-terminal of LacI for FROS imaging in
            B. subtilis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8297)
  AUTHORS   Altinoglu I,  Carballido-Lopez R
  TITLE     New PALM-compatible integration vectors for use in the Gram-positive
            model bacterium Bacillus subtilis
  JOURNAL   bioRxiv 2023
REFERENCE   2  (bases 1 to 8297)
  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"; journalName: "bioRxiv 
            2023"
COMMENT     Sequence Label: pIA035
FEATURES             Location/Qualifiers
     source          1..8297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1066..1085)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2103..2777
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     primer_bind     complement(3924..3943)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(6468..7056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6548..6567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7227..8018,8019..8087))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7850..7869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8088..8192)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8260..8278
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aacaaaattc tccagtcttc acatcggttt gaaaggagga agcggaagaa tgaagtaaga
       61 gggatttttg actccgaagt aagtcttcaa aaaatcaaat aaggagtgtc aagaatgttt
      121 gcaaaacgat tcaaaacctc tttactgccg ttattcgctg gatttttatt gctgtttcat
      181 ttggttctgg caggaccggc ggctgcgagt gctgaaacgg cgaacaaatc gaatgagctt
      241 acagcaccgt cgatcaaaag cggaaccatt cttcatgcat ggaattggtc gttcaatacg
      301 ttaaaacaca atatgaagga tattcatgat gcaggatata cagccattca gacatctccg
      361 attaaccaag taaaggaagg gaatcaagga gataaaagca tgtcgaactg gtactggctg
      421 tatcagccga catcgtatca aattggcaac cgttacttag gtactgaaca agaatttaaa
      481 gaaatgtgtg cagccgctga agaatatggc ataaaggtca ttgttgacgc ggtcatcaat
      541 cataccacca gtgattatgc cgcgatttcc aatgaggtta agagtattcc aaactggaca
      601 catggaaaca cacaaattaa aaactggtct gatcgatggg atgtcacgca gaattaaata
      661 gtacataatg gatttcctta cgcgaaatac gggcagacat ggcctgcccg gttattatta
      721 tttttgacac cagaccaact ggtaatggta gcgaccggcg ctcaggatcg ctagcgaatt
      781 cgagctcggt acggggacgg gttatcgaat tcccggtgga aacgaggtca tcatttcctt
      841 ccgaaaaaac ggttgcattt aaatcttaca tatgtaatac tttcaaagac tacatttgta
      901 agatttgatg tttgagtcgg ctgaaagatc gtacgtacca attattgttt cgtgattgtt
      961 caagccataa cactgtaggg atagtggaaa gagtgcttca tctggttacg atcaatcaaa
     1021 tattcaaacg gagggagacg attttgatga aaccagtaac gttatacgat gtcgcagagt
     1081 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
     1141 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
     1201 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
     1261 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
     1321 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa acggcggtgc
     1381 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     1441 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     1501 accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
     1561 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     1621 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
     1681 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
     1741 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
     1801 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
     1861 gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg
     1921 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
     1981 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc
     2041 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcggccg
     2101 ccatgagtgt gattaaacca gacatgaaga tcaagctgcg tatggaaggc gctgtaaatg
     2161 gacacccgtt cgcgattgaa ggagttggcc ttgggaagcc tttcgaggga aaacagagta
     2221 tggaccttaa agtcaaagaa ggcggacctc tgcctttcgc ctatgacatc ttgacaactg
     2281 tgttctgtta cggcaacagg gtattcgcca aatacccaga aaatatagta gactatttca
     2341 agcagtcgtt tcctgagggc tactcttggg aacgaagcat gaattacgaa gacgggggca
     2401 tttgtaacgc gacaaacgac ataaccctgg atggtgactg ttatatctat gaaattcgat
     2461 ttgatggtgt gaactttcct gccaatggtc cagttatgca gaagaggact gtgaaatggg
     2521 agccatccac tgagaaattg tatgtgcgtg atggagtgct gaagggtgat gttaacatgg
     2581 ctctgtcgct tgaaggaggt ggccattacc gatgtgactt caaaactact tataaagcta
     2641 agaaggttgt ccagttgcca gactatcact ttgtggacca ccacattgag attaaaagcc
     2701 acgacaaaga ttacagtaat gttaatctgc atgagcatgc cgaagcgcat tctgagctgc
     2761 cgaggcaggc caagtaaacc tgcaggcatg caagcttggc tgtaattctc atgtttgaca
     2821 gcttatcatc ggcaatagtt acccttatta tcaagataag aaagaaaagg atttttcgct
     2881 acgctcaaat cctttaaaaa aacacaaaag accacatttt ttaatgtggt cttttattct
     2941 tcaactaaag cacccattag ttcaacaaac gaaaattgga taaagtggga tatttttaaa
     3001 atatatattt atgttacagt aatattgact tttaaaaaag gattgattct aatgaagaaa
     3061 gcagacaagt aagcctccta aattcacttt agataaaaat ttaggaggca tatcaaatga
     3121 actttaataa aattgattta gacaattgga agagaaaaga gatatttaat cattatttga
     3181 accaacaaac gacttttagt ataaccacag aaattgatat tagtgtttta taccgaaaca
     3241 taaaacaaga aggatataaa ttttaccctg catttatttt cttagtgaca agggtgataa
     3301 actcaaatac agcttttaga actggttaca atagcgacgg agagttaggt tattgggata
     3361 agttagagcc actttataca atttttgatg gtgtatctaa aacattctct ggtatttgga
     3421 ctcctgtaaa gaatgacttc aaagagtttt atgatttata cctttctgat gtagagaaat
     3481 ataatggttc ggggaaattg tttcccaaaa cacctatacc tgaaaatgct ttttctcttt
     3541 ctattattcc atggacttca tttactgggt ttaacttaaa tatcaataat aatagtaatt
     3601 accttctacc cattattaca gcaggaaaat tcattaataa aggtaattca atatatttac
     3661 cgctatcttt acaggtacat cattctgttt gtgatggtta tcatgcagga ttgtttatga
     3721 actctattca ggaattgtca gataggccta atgactggct tttataatat gagataatgc
     3781 cgactgtact ttttacagtc ggttttctaa tgtcactaac ctgccccgtt agttgaagaa
     3841 ggtttttata ttacagctcc agatcctcta cgccggacgc atcgtggccg gcatcaccgg
     3901 cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc
     3961 tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc
     4021 cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa
     4081 cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg
     4141 acatggatga gcgatgatga tatccgttta ggctgggcgg tgatagcttc tcgttcaggc
     4201 agtacgcctc ttttcttttc cagacctgag ggaggcggaa atggtgtgag gttcccgggg
     4261 aaaagccaaa taggcgatcg cgggagtgct ttatttgaag atcaggctat cactgcggtc
     4321 aatagatttc acaatgtgat ggctggacag cctgaggaac tctcgaaccc gaatggaaac
     4381 aaccagatat ttatgaatca gcgcggctca catggcgttg tgctggcaaa tgcaggttca
     4441 tcctctgtct ctatcaatac ggcaacaaaa ttgcctgatg gcaggtatga caataaagct
     4501 ggagcgggtt catttcaagt gaacgatggt aaactgacag gcacgatcaa tgccaggtct
     4561 gtagctgtgc tttatcctga tgatattgca aaagcgcctc atgttttcct tgagaattac
     4621 aaaacaggtg taacacattc tttcaatgat caactgacga ttaccttgcg tgcagatgcg
     4681 aatacaacaa aagccgttta tcaaatcaat aatggaccag agacggcgtt taaggatgga
     4741 gatcaattca caatcggaaa aggagatcca tttggcaaaa catacaccat catgttaaaa
     4801 ggaacgaaca gtgatggtgt aacgaggacc gagaaataca gttttgttaa aagagatcca
     4861 gcgtcggcca aaaccatcgg ctatcaaaat ccgaatcatt ggagccaggt aaatgcttat
     4921 atctataaac atgatgggag ccgagtaatt gaattgaccg gatcttggcc tggaaaacca
     4981 atgactaaaa atgcagacgg aatttacacg ctgacgctgc ctgcggacac ggatacaacc
     5041 aacgcaaaag tgatttttaa taatggcagc gcccaagtgc ccggtcagaa tcagcctggc
     5101 tttgattacg tgctaaatgg tttatataat gactcgggct taagcggttc tcttccccat
     5161 tgagggcaag gctagacggg acttaccgaa agaaaccatc aatgatggtt tcttttttgt
     5221 tcataaatca gacaaaactt ttctcttgca aaagtttgtg aagtgttgca caatataaat
     5281 gtgaaatact tcacaaacaa aaagacatca aagagaaaca taccctgcaa ggatgctgat
     5341 attgtctgca tttgcgccgg agcaaaccaa aaacctggtg agacacgcct tgaattagta
     5401 gaaaagaact tgaagatttt caaaggcatc gttagtgaag tcatggcgag cggatttgac
     5461 ggcattttct tagtcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
     5521 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
     5581 catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga
     5641 ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg
     5701 attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
     5761 cgggccacct actgaagtgg atttctttaa gagctccttt aacttcctca ccagtagttg
     5821 tatcggtacc ataagtagaa gcagcaaccc aagtagcttt accagcatcc ggttcaacca
     5881 gcatagtaag aatcttactg gacatcggca gttcttcgaa cagtgcgcca actaccagct
     5941 ctttctgcag ttcattcagg gcaccggaga acctgcgtgc aatccatctt gttcaatcat
     6001 gcgaaacgat cctcatcctg tctcttgatc catggattac gcgttaaccc gggcccgcgg
     6061 atgcatatga tcagatctta aggcctaggt ctagagtctt tgttttgacg ccattagcgt
     6121 acgtaacaat cctcgttaaa ggacaaggac ctgagcggaa gtgtatcgta cagtagacgg
     6181 agtatactag tatagtctat agtccgtgga attattatat ttatctccga cgatattctc
     6241 atcagtgaaa tccagctgga gttctttagc aaattttttt attagctgaa cttagtatta
     6301 gtggccatac tcctccaatc caaagctatt tagaaagatt actatatcct caaacaggcg
     6361 gtaaccggcc tcttcatcgg gaatgcgcgc gaccttcagc atcgccggca tgtccccctg
     6421 gcggacggga agtatccagc tcgaggtcgg gccgcgttgc tggcgttttt ccataggctc
     6481 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6541 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6601 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6661 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6721 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6781 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6841 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6901 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6961 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7021 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7081 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7141 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7201 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7261 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7321 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7381 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7441 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7501 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     7561 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7621 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7681 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7741 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7801 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     7861 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7921 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7981 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8041 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8101 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8161 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8221 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8281 tttcgtcttc aagaatt
//
