LOCUS       sequence-443100-        8298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  amyE::(Ppen-lacI-Delta-11-bsdronPA cm). PALM integration vector 
            includes codon optimized 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 8298)
  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 8298)
  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: pIA036
FEATURES             Location/Qualifiers
     source          1..8298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1067..1086)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(3925..3944)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(6469..7057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6549..6568)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7228..8019,8020..8088))
                     /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     7851..7870
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8089..8193)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8261..8279
                     /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 acggggatcg ggttatcgaa ttcccggtgg aaacgaggtc atcatttcct
      841 tccgaaaaaa cggttgcatt taaatcttac atatgtaata ctttcaaaga ctacatttgt
      901 aagatttgat gtttgagtcg gctgaaagat cgtacgtacc aattattgtt tcgtgattgt
      961 tcaagccata acactgtagg gatagtggaa agagtgcttc atctggttac gatcaatcaa
     1021 atattcaaac ggagggagac gattttgatg aaaccagtaa cgttatacga tgtcgcagag
     1081 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     1141 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     1201 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
     1261 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
     1321 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa aacggcggtg
     1381 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
     1441 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
     1501 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
     1561 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
     1621 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
     1681 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
     1741 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
     1801 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     1861 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
     1921 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     1981 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
     2041 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcggcc
     2101 gccatgtcag tcataaaacc ggatatgaaa atcaaactcc gtatggaagg tgctgttaat
     2161 ggccatcctt tcgctattga aggagttgga ttgggcaaac ctttcgaagg caaacaatcc
     2221 atggatttga aggtgaaaga aggaggtccg ttaccatttg cgtatgacat tctgacaacg
     2281 gtgttttgct atggcaatcg agtgtttgcc aaatatcccg aaaacatcgt tgattacttc
     2341 aaacagagct ttcctgaagg gtattcatgg gaacgctcga tgaactatga agatggtggc
     2401 atttgtaatg cgacaaatga cataactctg gatggcgact gctatatcta cgagattcgc
     2461 tttgatggtg tcaattttcc ggcaaatgga ccagttatgc agaaaaggac cgtgaaatgg
     2521 gagccttcta cagagaagct gtatgtcaga gatggggtac ttaaaggcga cgtaaacatg
     2581 gcattatccc ttgaaggagg agggcattac cggtgtgact tcaaaaccac gtataaagcc
     2641 aagaaggtag ttcaactgcc ggattaccac tttgtcgatc atcacattga gatcaagagt
     2701 catgacaagg attacagcaa cgtgaacctt catgaacacg ctgaagcaca ttctgagtta
     2761 ccgagacaag cgaaataaac ctgcaggcat gcaagcttgg ctgtaattct catgtttgac
     2821 agcttatcat cggcaatagt tacccttatt atcaagataa gaaagaaaag gatttttcgc
     2881 tacgctcaaa tcctttaaaa aaacacaaaa gaccacattt tttaatgtgg tcttttattc
     2941 ttcaactaaa gcacccatta gttcaacaaa cgaaaattgg ataaagtggg atatttttaa
     3001 aatatatatt tatgttacag taatattgac ttttaaaaaa ggattgattc taatgaagaa
     3061 agcagacaag taagcctcct aaattcactt tagataaaaa tttaggaggc atatcaaatg
     3121 aactttaata aaattgattt agacaattgg aagagaaaag agatatttaa tcattatttg
     3181 aaccaacaaa cgacttttag tataaccaca gaaattgata ttagtgtttt ataccgaaac
     3241 ataaaacaag aaggatataa attttaccct gcatttattt tcttagtgac aagggtgata
     3301 aactcaaata cagcttttag aactggttac aatagcgacg gagagttagg ttattgggat
     3361 aagttagagc cactttatac aatttttgat ggtgtatcta aaacattctc tggtatttgg
     3421 actcctgtaa agaatgactt caaagagttt tatgatttat acctttctga tgtagagaaa
     3481 tataatggtt cggggaaatt gtttcccaaa acacctatac ctgaaaatgc tttttctctt
     3541 tctattattc catggacttc atttactggg tttaacttaa atatcaataa taatagtaat
     3601 taccttctac ccattattac agcaggaaaa ttcattaata aaggtaattc aatatattta
     3661 ccgctatctt tacaggtaca tcattctgtt tgtgatggtt atcatgcagg attgtttatg
     3721 aactctattc aggaattgtc agataggcct aatgactggc ttttataata tgagataatg
     3781 ccgactgtac tttttacagt cggttttcta atgtcactaa cctgccccgt tagttgaaga
     3841 aggtttttat attacagctc cagatcctct acgccggacg catcgtggcc ggcatcaccg
     3901 gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg
     3961 ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg
     4021 ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca
     4081 acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc
     4141 gacatggatg agcgatgatg atatccgttt aggctgggcg gtgatagctt ctcgttcagg
     4201 cagtacgcct cttttctttt ccagacctga gggaggcgga aatggtgtga ggttcccggg
     4261 gaaaagccaa ataggcgatc gcgggagtgc tttatttgaa gatcaggcta tcactgcggt
     4321 caatagattt cacaatgtga tggctggaca gcctgaggaa ctctcgaacc cgaatggaaa
     4381 caaccagata tttatgaatc agcgcggctc acatggcgtt gtgctggcaa atgcaggttc
     4441 atcctctgtc tctatcaata cggcaacaaa attgcctgat ggcaggtatg acaataaagc
     4501 tggagcgggt tcatttcaag tgaacgatgg taaactgaca ggcacgatca atgccaggtc
     4561 tgtagctgtg ctttatcctg atgatattgc aaaagcgcct catgttttcc ttgagaatta
     4621 caaaacaggt gtaacacatt ctttcaatga tcaactgacg attaccttgc gtgcagatgc
     4681 gaatacaaca aaagccgttt atcaaatcaa taatggacca gagacggcgt ttaaggatgg
     4741 agatcaattc acaatcggaa aaggagatcc atttggcaaa acatacacca tcatgttaaa
     4801 aggaacgaac agtgatggtg taacgaggac cgagaaatac agttttgtta aaagagatcc
     4861 agcgtcggcc aaaaccatcg gctatcaaaa tccgaatcat tggagccagg taaatgctta
     4921 tatctataaa catgatggga gccgagtaat tgaattgacc ggatcttggc ctggaaaacc
     4981 aatgactaaa aatgcagacg gaatttacac gctgacgctg cctgcggaca cggatacaac
     5041 caacgcaaaa gtgattttta ataatggcag cgcccaagtg cccggtcaga atcagcctgg
     5101 ctttgattac gtgctaaatg gtttatataa tgactcgggc ttaagcggtt ctcttcccca
     5161 ttgagggcaa ggctagacgg gacttaccga aagaaaccat caatgatggt ttcttttttg
     5221 ttcataaatc agacaaaact tttctcttgc aaaagtttgt gaagtgttgc acaatataaa
     5281 tgtgaaatac ttcacaaaca aaaagacatc aaagagaaac ataccctgca aggatgctga
     5341 tattgtctgc atttgcgccg gagcaaacca aaaacctggt gagacacgcc ttgaattagt
     5401 agaaaagaac ttgaagattt tcaaaggcat cgttagtgaa gtcatggcga gcggatttga
     5461 cggcattttc ttagtcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc
     5521 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
     5581 ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcattgg
     5641 accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg
     5701 gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
     5761 ccgggccacc tactgaagtg gatttcttta agagctcctt taacttcctc accagtagtt
     5821 gtatcggtac cataagtaga agcagcaacc caagtagctt taccagcatc cggttcaacc
     5881 agcatagtaa gaatcttact ggacatcggc agttcttcga acagtgcgcc aactaccagc
     5941 tctttctgca gttcattcag ggcaccggag aacctgcgtg caatccatct tgttcaatca
     6001 tgcgaaacga tcctcatcct gtctcttgat ccatggatta cgcgttaacc cgggcccgcg
     6061 gatgcatatg atcagatctt aaggcctagg tctagagtct ttgttttgac gccattagcg
     6121 tacgtaacaa tcctcgttaa aggacaagga cctgagcgga agtgtatcgt acagtagacg
     6181 gagtatacta gtatagtcta tagtccgtgg aattattata tttatctccg acgatattct
     6241 catcagtgaa atccagctgg agttctttag caaatttttt tattagctga acttagtatt
     6301 agtggccata ctcctccaat ccaaagctat ttagaaagat tactatatcc tcaaacaggc
     6361 ggtaaccggc ctcttcatcg ggaatgcgcg cgaccttcag catcgccggc atgtccccct
     6421 ggcggacggg aagtatccag ctcgaggtcg ggccgcgttg ctggcgtttt tccataggct
     6481 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6541 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6601 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6661 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6721 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6781 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6841 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6901 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6961 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7021 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7081 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7141 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7201 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7261 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7321 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7381 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7441 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7501 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc
     7561 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7621 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7681 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7741 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7801 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc
     7861 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7921 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7981 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8041 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8101 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8161 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8221 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8281 ctttcgtctt caagaatt
//
