LOCUS       sequence-405455-        7005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  amyE::(dronPA cm), for dronPA fusions. PALM-compatible integration 
            vector for Bacillus subtilis. Includes dronPA photoswitchable FP 
            with multi cloning sites (MCS) for fast and interchangeable 
            cloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7005)
  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 7005)
  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: pIA027
FEATURES             Location/Qualifiers
     source          1..7005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             811..1485
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     primer_bind     complement(2632..2651)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(5176..5764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5256..5275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5935..6726,6727..6795))
                     /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     6558..6577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6796..6900)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6968..6986
                     /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 acggggatcc tgcggccgcc atgagtgtga ttaaaccaga catgaagatc
      841 aagctgcgta tggaaggcgc tgtaaatgga cacccgttcg cgattgaagg agttggcctt
      901 gggaagcctt tcgagggaaa acagagtatg gaccttaaag tcaaagaagg cggacctctg
      961 cctttcgcct atgacatctt gacaactgtg ttctgttacg gcaacagggt attcgccaaa
     1021 tacccagaaa atatagtaga ctatttcaag cagtcgtttc ctgagggcta ctcttgggaa
     1081 cgaagcatga attacgaaga cgggggcatt tgtaacgcga caaacgacat aaccctggat
     1141 ggtgactgtt atatctatga aattcgattt gatggtgtga actttcctgc caatggtcca
     1201 gttatgcaga agaggactgt gaaatgggag ccatccactg agaaattgta tgtgcgtgat
     1261 ggagtgctga agggtgatgt taacatggct ctgtcgcttg aaggaggtgg ccattaccga
     1321 tgtgacttca aaactactta taaagctaag aaggttgtcc agttgccaga ctatcacttt
     1381 gtggaccacc acattgagat taaaagccac gacaaagatt acagtaatgt taatctgcat
     1441 gagcatgccg aagcgcattc tgagctgccg aggcaggcca agtaaacctg caggcatgca
     1501 agcttggctg taattctcat gtttgacagc ttatcatcgg caatagttac ccttattatc
     1561 aagataagaa agaaaaggat ttttcgctac gctcaaatcc tttaaaaaaa cacaaaagac
     1621 cacatttttt aatgtggtct tttattcttc aactaaagca cccattagtt caacaaacga
     1681 aaattggata aagtgggata tttttaaaat atatatttat gttacagtaa tattgacttt
     1741 taaaaaagga ttgattctaa tgaagaaagc agacaagtaa gcctcctaaa ttcactttag
     1801 ataaaaattt aggaggcata tcaaatgaac tttaataaaa ttgatttaga caattggaag
     1861 agaaaagaga tatttaatca ttatttgaac caacaaacga cttttagtat aaccacagaa
     1921 attgatatta gtgttttata ccgaaacata aaacaagaag gatataaatt ttaccctgca
     1981 tttattttct tagtgacaag ggtgataaac tcaaatacag cttttagaac tggttacaat
     2041 agcgacggag agttaggtta ttgggataag ttagagccac tttatacaat ttttgatggt
     2101 gtatctaaaa cattctctgg tatttggact cctgtaaaga atgacttcaa agagttttat
     2161 gatttatacc tttctgatgt agagaaatat aatggttcgg ggaaattgtt tcccaaaaca
     2221 cctatacctg aaaatgcttt ttctctttct attattccat ggacttcatt tactgggttt
     2281 aacttaaata tcaataataa tagtaattac cttctaccca ttattacagc aggaaaattc
     2341 attaataaag gtaattcaat atatttaccg ctatctttac aggtacatca ttctgtttgt
     2401 gatggttatc atgcaggatt gtttatgaac tctattcagg aattgtcaga taggcctaat
     2461 gactggcttt tataatatga gataatgccg actgtacttt ttacagtcgg ttttctaatg
     2521 tcactaacct gccccgttag ttgaagaagg tttttatatt acagctccag atcctctacg
     2581 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     2641 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
     2701 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
     2761 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
     2821 tgcaggagtc gcataaggga gagcgtcgac atggatgagc gatgatgata tccgtttagg
     2881 ctgggcggtg atagcttctc gttcaggcag tacgcctctt ttcttttcca gacctgaggg
     2941 aggcggaaat ggtgtgaggt tcccggggaa aagccaaata ggcgatcgcg ggagtgcttt
     3001 atttgaagat caggctatca ctgcggtcaa tagatttcac aatgtgatgg ctggacagcc
     3061 tgaggaactc tcgaacccga atggaaacaa ccagatattt atgaatcagc gcggctcaca
     3121 tggcgttgtg ctggcaaatg caggttcatc ctctgtctct atcaatacgg caacaaaatt
     3181 gcctgatggc aggtatgaca ataaagctgg agcgggttca tttcaagtga acgatggtaa
     3241 actgacaggc acgatcaatg ccaggtctgt agctgtgctt tatcctgatg atattgcaaa
     3301 agcgcctcat gttttccttg agaattacaa aacaggtgta acacattctt tcaatgatca
     3361 actgacgatt accttgcgtg cagatgcgaa tacaacaaaa gccgtttatc aaatcaataa
     3421 tggaccagag acggcgttta aggatggaga tcaattcaca atcggaaaag gagatccatt
     3481 tggcaaaaca tacaccatca tgttaaaagg aacgaacagt gatggtgtaa cgaggaccga
     3541 gaaatacagt tttgttaaaa gagatccagc gtcggccaaa accatcggct atcaaaatcc
     3601 gaatcattgg agccaggtaa atgcttatat ctataaacat gatgggagcc gagtaattga
     3661 attgaccgga tcttggcctg gaaaaccaat gactaaaaat gcagacggaa tttacacgct
     3721 gacgctgcct gcggacacgg atacaaccaa cgcaaaagtg atttttaata atggcagcgc
     3781 ccaagtgccc ggtcagaatc agcctggctt tgattacgtg ctaaatggtt tatataatga
     3841 ctcgggctta agcggttctc ttccccattg agggcaaggc tagacgggac ttaccgaaag
     3901 aaaccatcaa tgatggtttc ttttttgttc ataaatcaga caaaactttt ctcttgcaaa
     3961 agtttgtgaa gtgttgcaca atataaatgt gaaatacttc acaaacaaaa agacatcaaa
     4021 gagaaacata ccctgcaagg atgctgatat tgtctgcatt tgcgccggag caaaccaaaa
     4081 acctggtgag acacgccttg aattagtaga aaagaacttg aagattttca aaggcatcgt
     4141 tagtgaagtc atggcgagcg gatttgacgg cattttctta gtcgcgacgc gaggctggat
     4201 ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc
     4261 catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc
     4321 tcttaccagc ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc
     4381 ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct
     4441 ccccgcgttg cgtcgcggtg catggagccg ggccacctac tgaagtggat ttctttaaga
     4501 gctcctttaa cttcctcacc agtagttgta tcggtaccat aagtagaagc agcaacccaa
     4561 gtagctttac cagcatccgg ttcaaccagc atagtaagaa tcttactgga catcggcagt
     4621 tcttcgaaca gtgcgccaac taccagctct ttctgcagtt cattcagggc accggagaac
     4681 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcca
     4741 tggattacgc gttaacccgg gcccgcggat gcatatgatc agatcttaag gcctaggtct
     4801 agagtctttg ttttgacgcc attagcgtac gtaacaatcc tcgttaaagg acaaggacct
     4861 gagcggaagt gtatcgtaca gtagacggag tatactagta tagtctatag tccgtggaat
     4921 tattatattt atctccgacg atattctcat cagtgaaatc cagctggagt tctttagcaa
     4981 atttttttat tagctgaact tagtattagt ggccatactc ctccaatcca aagctattta
     5041 gaaagattac tatatcctca aacaggcggt aaccggcctc ttcatcggga atgcgcgcga
     5101 ccttcagcat cgccggcatg tccccctggc ggacgggaag tatccagctc gaggtcgggc
     5161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5581 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5881 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5941 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6001 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6061 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6121 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6181 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6241 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6301 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6361 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6421 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6481 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6541 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6601 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6661 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6721 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6781 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6841 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6901 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6961 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaatt
//
