LOCUS       sequence-405349-        8104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  thrC::(Ppen-lacI-Delta-11-PAmCherry1 erm2). PALM integration vector 
            includes PAmCherry1 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 8104)
  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 8104)
  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: pIA030
FEATURES             Location/Qualifiers
     source          1..8104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1050..1069)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2087..2794
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein"
                     /label=PAmCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALKG
                     EVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2229..2247)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2497..2516
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     rep_origin      complement(6275..6863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6355..6374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7034..7825,7826..7894))
                     /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     7657..7676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7895..7999)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8067..8085
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aattcatgta aaagatgagg ttggttcatt ctcgaaaatt acatctgtgt tctcagagcg
       61 gggcgtgagc tttgaaaaaa tccttcagct gccaattaaa ggccatgatg agttagctga
      121 aatcgtaatt gtcacacatc atacatcaga agctgatttc agtgatatcc tgcaaaacct
      181 aaatgatttg gaagtcgttc aagaagtcaa aagcacatat cgtgtagaag ggaacggttg
      241 gagctaatgt ggaaaggact tatccatcaa tataaagaat ttttacctgt aacagatcaa
      301 acaccggcgc taactttaca tgaaggaaac acacctctta ttcacctgcc gaagctgtct
      361 gagcagctcg gaattgagct tcatgtcaaa acggaaggcg tcaatcctac gggatcattt
      421 aaagatcgcg gaatggttat ggctgtggca aaggcaaaag aagaaggcaa tgacacgatt
      481 atgtgcgcgt caacaggtaa cacttccgct gctgcggcag catatgcagc ccgtgctaac
      541 atgaaatgca ttgtcatcat cccgaacgga aaaattgcat ttggaaaact cgctcaagct
      601 gtcatgtacg gagccgagat tatcgcaatt gacggaaact ttgacgatgc gcttaaaatt
      661 gtccgttcca tctgtgagaa atcaccgatt gcccttgtca actcagtcaa cccttaccgc
      721 attgaaggcc aaaaaactgc tgccttcgga tcgctagcga attcgagctc ggtacgggga
      781 tcgggttatc gaattcccgg tggaaacgag gtcatcattt ccttccgaaa aaacggttgc
      841 atttaaatct tacatatgta atactttcaa agactacatt tgtaagattt gatgtttgag
      901 tcggctgaaa gatcgtacgt accaattatt gtttcgtgat tgttcaagcc ataacactgt
      961 agggatagtg gaaagagtgc ttcatctggt tacgatcaat caaatattca aacggaggga
     1021 gacgattttg atgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
     1081 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
     1141 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
     1201 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
     1261 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
     1321 gatggtagaa cgaagcggcg tcgaagcctg taaaacggcg gtgcacaatc ttctcgcgca
     1381 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
     1441 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
     1501 cagcattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
     1561 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
     1621 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
     1681 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
     1741 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
     1801 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     1861 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     1921 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     1981 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     2041 ctctccccgc gcgttggccg attcattaat gcagctggcg gccgccgtga gcaagggcga
     2101 ggaggataac atggccatca ttaaggagtt catgcgcttc aaggtgcaca tggaggggtc
     2161 cgtgaacggc cacgtgttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     2221 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcacct gggacatcct
     2281 gtcccctcaa ttcatgtacg gctccaatgc ctacgtgaag caccccgccg acatccccga
     2341 ctactttaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga aattcgagga
     2401 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggtgagt tcatctacaa
     2461 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     2521 gggctgggag gccctctccg agcggatgta ccccgaggac ggcgccctga agggcgaggt
     2581 caagccgaga gtgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     2641 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaaccgca agttggacat
     2701 cacctcacac aacgaggact acaccatcgt ggaacagtac gaacgtgccg agggccgcca
     2761 ctccaccggc ggcatggacg agctgtacaa gtaaaggcat gcaagcttgg ctgtaattcg
     2821 ataaccctaa agttatggaa ataagactta gaagcaaact taagagtgtg ttgatagtgc
     2881 attatcttaa aattttgtat aataggaatt gaagttaaat tagatgctaa aaatttgtaa
     2941 ttaagaagga gggattcgtc atgttggtat tccaaatgcg taatgtagat aaaacatcta
     3001 ctgttttgaa acagactaaa aacagtgatt acgcagataa ataaatacgt tagattaatt
     3061 cctaccagtg actaatctta tgacttttta aacagataac taaaattaca aacaaatcgt
     3121 ttaacttctg tatttattta tagatgtaat cacttcagga gtgattacat gaacaaaaat
     3181 ataaaatatt ctcaaaactt tttaacgagt gaaaaagtac tcaaccaaat aataaaacaa
     3241 ttgaatttaa aagaaaccga taccgtttac gaaattggaa caggtaaagg gcatttaacg
     3301 acgaaactgg ctaaaataag taaacaggta acgtctattg aattagacag tcatctattc
     3361 aacttatcgt cagaaaaatt aaaactgaac attcgtgtca ctttaattca ccaagatatt
     3421 ctacagtttc aattccctaa caaacagagg tataaaattg ttgggaatat tccttaccat
     3481 ttaagcacac aaattattaa aaaagtggtt tttgaaagcc atgcgtctga catctatctg
     3541 attgttgaag aaggattcta caagcgtacc ttggatattc accgaacact agggttgctc
     3601 ttgcacactc aagtctcgat tcagcaattg cttaagctgc cagcggaatg ctttcatcct
     3661 aaaccaaaag taaacagtgt cttaataaaa cttacccgcc ataccacaga tgttccagat
     3721 aaatattgga agctatatac gtactttgtt tcaaaatggg tcaatcgaga atatcgtcaa
     3781 ctgtttacta aaaatcagtt tcatcaagca atgaaacacg ccaaagtaaa caatttaagt
     3841 accgttactt atgagcaagt attgtctatt tttaatagtt atctattatt taacgggagg
     3901 aaataattct atgagtcgct tttgtaaatt tggaaagtta cacgttacta aagggaatgt
     3961 agataaatta ttaggtatac tactgacagc ttccaaggag ctaaagaggt ccctagcgcc
     4021 tacggggaat ttgtatcgaa gtgtgcgaac agctggggga agcgcctgat gttttagcga
     4081 tcccggtcgg aaatgctgga aacatcactg cgtactggaa gggcttcaag gaatatcatg
     4141 agaaaaacgg cacaggcctt ccgaaaatgc gcggctttga agctgagggt gcggcggcaa
     4201 tcgtgcgcaa tgaagtgatt gaaaatccgg aaacaatagc gacagccatt cgtatcggaa
     4261 acccggcaag ctgggacaaa gctgtaaagg cagccgagga atccaatggg aaaattgacg
     4321 aagtcactga tgatgaaatc cttcacgcat atcagctgat cgcccgtgta gaaggcgtgt
     4381 ttgcagaacc aggttcttgc gcgtctatcg caggagtgct gaaacaggtg aaatccggag
     4441 aaattccgaa aggcagcaag gtcgtagctg tgttaacagg aaacggactg aaagatccga
     4501 acacagcggt cgacatttca gaaatcaagc ctgtcacatt gccgactgat gaagacagca
     4561 tccttgaata tgtaaaagga gcggcccgtg tatgaacgaa gccgacatgc tgttctctgt
     4621 cactgttccc ggaagcacag ctaacctagg ccccggcttt gattcagtcg gaatggcgct
     4681 cagcagatat ttgaagctga ccgtctttga aagcgacaaa tggtcttttg aggctgaaac
     4741 agaaacagtc gccggaatta attcgagctc ctttaacttc ctcaccagta gttgtatcgg
     4801 taccataagt agaagcagca acccaagtag ctttaccagc atccggttca accagcatag
     4861 taagaatctt actggacatc ggcagttctt cgaacagtgc gccaactacc agctctttct
     4921 gcagttcatt cagggcaccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
     4981 cgatcctcat cctgtctctt gatccatgga ttacgcgtta acccgggccc gcggatgcat
     5041 atgatcagat cttaaggcct aggtctagag gatcgatctg tataataaag aataattatt
     5101 aatctgtaga caaattgtga aaggatgtac ttaaacgcta acggtcagct ttattgaaca
     5161 gtaatttaag tatatgtcca atctagggta agtaaattga gtatcaatat aaactttata
     5221 tgaacataat caacgaggtg aaatcatgag caatttgatt aacggaaaaa taccaaatca
     5281 agcgattcaa acattaaaaa tcgtaaaaga tttatttgga agttcaatag ttggagtata
     5341 tctatttggt tcagcagtaa atggtggttt acgcattaac agcgatgtag atgttctagt
     5401 cgtcgtgaat catagtttac ctcaattaac tcgaaaaaaa ctaacagaaa gactaatgac
     5461 tatatcagga aagattggaa atacggattc tgttagacca cttgaagtta cggttataaa
     5521 taggagtgaa gttgtccctt ggcaatatcc tccaaaaaga gaatttatat acggtgagtg
     5581 gctcaggggt gaatttgaga atggacaaat tcaggaacca agctatgatc ctgatttggc
     5641 tattgtttta gcacaagcaa gaaagaatag tatttctcta tttggtcctg attcttcaag
     5701 tatacttgtc tccgtacctt tgacagatat tcgaagagca attaaggatt ctttgccaga
     5761 actaattgag gggataaaag gtgatgagcg taatgtaatt ttaaccctag ctcgaatgtg
     5821 gcaaacagtg actactggtg aaattacctc gaaagatgtc gctgcagaat gggctatacc
     5881 tcttttacct aaagagcatg taactttact ggatatagct agaaaaggct atcggggaga
     5941 gtgtgatgat aagtgggaag gactatattc aaaggtgaaa gcactcgtta agtatatgaa
     6001 aaattctata gaaacttctc tcaattaggc taattttatt gcaataacag gtgcttactt
     6061 ttctggagtt ctttagcaaa tttttttatt agctgaactt agtattagtg gccatactcc
     6121 tccaatccaa agctatttag aaagattact atatcctcaa acaggcggta accggcctct
     6181 tcatcgggaa tgcgcgcgac cttcagcatc gccggcatgt ccccctggcg gacgggaagt
     6241 atccagctcg aggtcgggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6301 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6361 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6421 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6481 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6541 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6601 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6661 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     6721 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6781 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6841 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6901 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6961 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7021 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7081 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7141 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7201 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7261 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7321 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     7381 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7441 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7501 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7561 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7621 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     7681 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7741 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7801 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7861 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7921 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7981 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     8041 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     8101 aatt
//
