LOCUS       sequence-375002-        4489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses blue chromoprotein on a broad host range RP4 origin 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4489)
  AUTHORS   Elston KM, Phillips LE, Leonard SP, Young E, Holley JC, Ahsanullah 
            T, McReynolds B, Moran NA, Barrick JE
  TITLE     The Pathfinder plasmid toolkit for genetically engineering newly 
            isolated bacteria enables the study of Drosophila-colonizing 
            Orbaceae.
  JOURNAL   ISME Commun. 2023 May 24;3(1):49. doi: 10.1038/s43705-023-00255-3.
  PUBMED    37225918
REFERENCE   2  (bases 1 to 4489)
  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: "ISME 
            Commun."; date: "2023-05-24"; volume: "3(1):49. doi"; pages: " 
            10.1038/s43705-023-00255-3"
COMMENT     Sequence Label: pBTK1010 (pSL25)
FEATURES             Location/Qualifiers
     source          1..4489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      259..790
                     /label=oriV
                     /note="incP origin of replication"
     CDS             1183..2331
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     oriT            2392..2500
                     /note="incP origin of transfer"
     primer_bind     complement(2807..2826)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2818..3633
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2818..3633
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2891..2910)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(3690..3717)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3733..3804)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atgtaatatc tcctctgttt ctgtataaca gtactatgtg attataccag ccccctcact
       61 acatgtcaag aataaactgc caaagctggt ctcaaggggc ctgtcagacc aagtttactc
      121 atatatactt tagattgatt tcagacagct gggcgcgccg tagaaaaaat caaaggatct
      181 tcttgagatc ctttttttct gcgggggatc aggaccgctg ccggagcgca acccactcac
      241 tacagcagag ccatgtaggg ccgccggcgt tgtggatacc tcgcggaaaa cttggccctc
      301 actgacagat gaggggcgga cgttgacact tgaggggccg actcacccgg cgcggcgttg
      361 acagatgagg ggcaggctcg atttcggccg gcgacgtgga gctggccagc ctcgcaaatc
      421 ggcgaaaacg cctgatttta cgcgagtttc ccacggatga tgtggacaag cctggggata
      481 agtgccctgc ggtattgaca cttgaggggc gcgactactg acagatgagg ggcgcgatcc
      541 ttgacacttg aggggcagag tgctgacaga tgaggggcgc acctattgac atttgagggg
      601 ctgtccacag gcagaaaatc cagcatttgc aagggtttcc gcccgttttt cggccaccgc
      661 taacctgtct tttaacctgc ttttaaacca atatttataa accttgtttt taaccagggc
      721 tgcgccctgt gcgcgtgacc gcgcacgccg aaggggggtg cccccccttc tcgaaccctc
      781 ccggcccgct aacgcgggcc tcccatcccc ccaggggctg cgcccctcgg ccgcgaacgg
      841 cctcacccca aaaatggcag ccacgtagaa agccagtccg cagaaacggt gctgaccccg
      901 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca
      961 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag
     1021 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
     1081 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat cgacggatcg
     1141 atccggggaa ttaattccgg ggcaatcccg caaggagggt gaatgaatcg gacgtttgac
     1201 cggaaggcat acaggcaaga actgatcgac gcggggtttt ccgccgagga tgccgaaacc
     1261 atcgcaagcc gcaccgtcat gcgtgcgccc cgcgaaacct tccagtccgt cggctcgata
     1321 gtccagcaag ctacggccaa gatcgagcgc gacagcgtgc aactggctcc ccctgccctg
     1381 cccgcgccat cggccgccgt ggagcgttcg cgtcgtctgg aacaggaggc ggcaggtttg
     1441 gcgaagtcga tgaccatcga cacgcgagga actatgacga ccaagaagcg aaaaaccgcc
     1501 ggcgaggacc tggcaaaaca ggtcagcgaa gccaagcagg ccgcgttgct gaaacacacg
     1561 aagcagcaga tcaaggaaat gcagctttcc ttgttcgata ttgcgccgtg gccggacacg
     1621 atgcgagcga tgccaaacga cacggcccgc tctgccctgt tcaccacgcg caacaagaaa
     1681 atcccgcgcg aggcgctgca aaacaaggtc attttccacg tcaacaagga cgtgaagatc
     1741 acctacaccg gcgtcgagct gcgggccgac gatgacgaac tggtgtggca gcaggtgttg
     1801 gagtacgcga agcgcacccc tatcggcgag ccgatcacct tcacgttcta cgagctttgc
     1861 caggacctgg gctggtcgat caatggccgg tattacacga aggccgagga atgcctgtcg
     1921 cgcctacagg cgacggcgat gggcttcacg tccgaccgcg ttgggcacct ggaatcggtg
     1981 tcgctgctgc accgcttccg cgtcctggac cgtggcaaga aaacgtcccg ttgccaggtc
     2041 ctgatcgacg aggaaatcgt cgtgctgttt gctggcgacc actacacgaa attcatatgg
     2101 gagaagtacc gcaagctgtc gccgacggcc cgacggatgt tcgactattt cagctcgcac
     2161 cgggagccgt acccgctcaa gctggaaacc ttccgcctca tgtgcggatc ggattccacc
     2221 cgcgtgaaga agtggcgcga gcaggtcggc gaagcctgcg aagagttgcg aggcagcggc
     2281 ctggtggaac acgcctgggt caatgatgac ctggtgcatt gcaaacgcta gggccttgtg
     2341 gggtcagttc cggctggggg ttcagcagcc acctgcatcg cggccggcct acggccagcc
     2401 tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga
     2461 acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca
     2521 ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg
     2581 atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcaa ggcggcaggc
     2641 tgactcacgt taagggattt tggtcatgac tgatccttca actcagcaaa agttcgattt
     2701 attcaacaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     2761 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     2821 agccatattc aacgggaaac gtcttgctcc aggccgcgat taaattccaa catggatgct
     2881 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     2941 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     3001 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     3061 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     3121 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     3181 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     3241 aacagcgatc gcgtatttcg cctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     3301 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     3361 atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     3421 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     3481 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     3541 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     3601 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     3661 gcggccgctc ggtgagaccg acttataaac gcagaaaggc ccacccgaag gtgagccagt
     3721 gtaactctag tagagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
     3781 cgactgagcc tttcgtttta tttgatgcct ggagatcctt ttattaggcg accacaggtt
     3841 tgcgtgcaat ggaaatttca cactgctcaa ccgaagtgta atccttgttg tgattggtta
     3901 catccagttt gcggtcaaca tagtgatacc ctggcatctt cacaggcttc tttgccttgt
     3961 aagtagtttt aaattcacac aaatagtgac cgcctccttc taacttcaga gccataaagt
     4021 tgtttcctag cagcattcca tctcgtgcaa agagacgctc agtgttgggt tcccagccct
     4081 gtgtcttctt ctgcatgaca ggtccattgg gaggaaagtt caaaccagag aacttgacat
     4141 ggtagatgaa acagttgcct tggatgctgg aatcattgct gacagtacac actgcaccat
     4201 cttcaaagtt catgatcctc tcccatgtat agccctccgg gaatgactgc tttacatagt
     4261 cagggatgtc tccagggtac ttggtgaatg gtatgcttcc gtactgacac tgtggtgata
     4321 aaatatccca agcaaatggc agaggtccgc ccttggtgac agtgagcttt accgtctgtt
     4381 ccccctcgta gggcttacct tttccatcgc cttcgacctc aaagtagtgt ccattgaccg
     4441 tgcctgacat ataaaccttg taggtcattt gtttagcgat cacactcat
//
