LOCUS       sequence-54251-e        4867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4867)
  AUTHORS   Blakeley BD, Chapman AM, McNaughton BR
  TITLE     Split-superpositive GFP reassembly is a fast, efficient, and robust 
            method for detecting protein-protein interactions in vivo.
  JOURNAL   Mol Biosyst. 2012 Aug;8(8):2036-40. Epub 2012 Jun 12.
  PUBMED    22692102
REFERENCE   2  (bases 1 to 4867)
  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: "Mol 
            Biosyst."; date: "2012-08"; volume: "8(8)"; pages: "2036-40. Epub 
            2012 Jun 12"
COMMENT     Sequence Label: pMRBad-Z-CspGFP
FEATURES             Location/Qualifiers
     source          1..4867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(427..445)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      441..488
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1090..1905)
                     /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             complement(1090..1905)
                     /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     1813..1832
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1897..1916
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2366..2383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2500..3044)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2580..2599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3624..4502)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4529..4813
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4736..4755
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4840..4862
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ccatggcaag cgagcagctg gaaaagaagt tacaagccct ggagaaaaaa cttgctcagc
       61 tggaatggaa aaaccaagca ttggaaaaaa aactcgcgca gacgtcgggt ggaagcggta
      121 agaatggtat caaggcaaaa ttcaaaattc gccataacgt gaaagacggc agcgttcaat
      181 tagcggatca ttatcaacaa aacacgccga ttggtcgcgg gcctgtactg ttacctcgca
      241 accactacct gagcacccgt tctaaactga gcaaagatcc gaaagaaaaa cgcgatcaca
      301 tggttctgtt agaattcgtg accgctgcag gcattaagca cggacgcgac gaacgctaca
      361 agtaatgtac aactaaggat ccggctgcta acaaagcccg aaaggaagct gagttggctg
      421 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
      481 gttttttgct gaaaggagga actatatccg gatatccaca ggacgggtgt ggtcgccatg
      541 atggtgacca ccatttgccc catatcaccg tcggtactga tcccgtcgtc aataaaccga
      601 accgctacac cctgagcatc aaactctttt atcagttgga tcatgtcggc ggtgtcgcgg
      661 ccaagacggt cgagcttctt caccagaatg acatcacctt cctccacctt catcctcagc
      721 aaatccagcc cttcccgatc tgttgaactg ccggatgcct tgtcggtaaa gatgcggtta
      781 gcttttaccc ctgcatcttt gagcgctgag gtctgcctcg tgaagaaggt gttgctgact
      841 cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca cggttgatga
      901 gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc acggaacggt
      961 ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cgatttattc
     1021 aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca accaattaac
     1081 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     1141 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     1201 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     1261 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     1321 agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc
     1381 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     1441 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     1501 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     1561 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     1621 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     1681 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     1741 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     1801 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     1861 taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     1921 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     1981 gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga
     2041 gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa
     2101 gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg gctccctcac
     2161 tttctggctg gatgatgggg cgattcaggc ctggtatgag tcagcaacac cttcttcacg
     2221 aggcagacct cagcgctagc ggagtgtata ctggcttact atgttggcac tgatgagggt
     2281 gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc accggtgcgt cagcagaata
     2341 tgtgatacag gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac
     2401 tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga
     2461 tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc
     2521 tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata
     2581 aagataccag gcgtttcccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
     2641 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
     2701 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
     2761 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
     2821 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
     2881 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
     2941 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
     3001 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
     3061 aggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3181 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3241 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3301 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3361 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3421 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3481 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcagat cgatgcataa
     3541 tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc cgtcaagccg
     3601 tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct
     3661 tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag
     3721 aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg
     3781 gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta
     3841 atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt
     3901 gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc
     3961 tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc
     4021 agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc
     4081 ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg
     4141 cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg
     4201 cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag
     4261 tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt
     4321 ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt
     4381 cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc
     4441 gccaccagat gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc
     4501 atacttttca tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat
     4561 tgccgtcact gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt
     4621 aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc
     4681 tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc
     4741 atagcatttt tatccataag attagcggat cctacctgac gctttttatc gcaactctct
     4801 actgtttctc catacccgtt tttttgggct agaaataatt ttgtttaact ttaagaagga
     4861 gatatac
//
