LOCUS       sequence-82268-e        4856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L-(+)-arabinose-inducible expression of Z-CGFP positive control for 
            in vivo split GFP assembly assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4856)
  AUTHORS   Magliery TJ, Wilson CG, Pan W, Mishler D, Ghosh I, Hamilton AD, 
            Regan L
  TITLE     Detecting protein-protein interactions with a green fluorescent 
            protein fragment reassembly trap: scope and mechanism.
  JOURNAL   J Am Chem Soc. 2005 Jan 12;127(1):146-57.
  PUBMED    15631464
REFERENCE   2  (bases 1 to 4856)
  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: "J Am Chem 
            Soc."; date: "2005-01-12"; volume: "127(1)"; pages: "146-57"
COMMENT     Sequence Label: pMRBAD-Z-CGFP
FEATURES             Location/Qualifiers
     source          1..4856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     302..322
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(416..434)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      430..477
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1079..1894)
                     /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(1079..1894)
                     /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     1802..1821
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1886..1905
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2355..2372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2489..3033)
                     /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(2569..2588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3613..4491)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4518..4802
                     /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     4725..4744
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4829..4851
                     /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 agaatggaat caaagtgaac ttcaagaccc gccacaacat tgaagatgga agcgttcaac
      181 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
      241 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
      301 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactgtaca
      361 actaaggatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
      421 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
      481 aaaggaggaa ctatatccgg atatccacag gacgggtgtg gtcgccatga tggtgaccac
      541 catttgcccc atatcaccgt cggtactgat cccgtcgtca ataaaccgaa ccgctacacc
      601 ctgagcatca aactctttta tcagttggat catgtcggcg gtgtcgcggc caagacggtc
      661 gagcttcttc accagaatga catcaccttc ctccaccttc atcctcagca aatccagccc
      721 ttcccgatct gttgaactgc cggatgcctt gtcggtaaag atgcggttag cttttacccc
      781 tgcatctttg agcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc
      841 tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg
      901 taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg
      961 ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc
     1021 cgtcccgtca agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt
     1081 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     1141 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     1201 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     1261 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     1321 aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc
     1381 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     1441 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     1501 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     1561 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     1621 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     1681 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     1741 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     1801 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     1861 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     1921 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
     1981 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     2041 agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca
     2101 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
     2161 atgatggggc gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc
     2221 agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt
     2281 gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg
     2341 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     2401 gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     2461 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     2521 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     2581 cgtttcccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt
     2641 cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg
     2701 cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct
     2761 tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag
     2821 cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa
     2881 actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag
     2941 ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag
     3001 caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa ggggtctgac
     3061 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3121 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3181 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3241 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3301 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3361 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3421 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3481 gttaatagtt tgcgcaacgt tgttgccatt gctgcagatc gatgcataat gtgcctgtca
     3541 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
     3601 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     3661 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     3721 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     3781 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     3841 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     3901 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     3961 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     4021 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     4081 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     4141 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     4201 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     4261 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     4321 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     4381 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     4441 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     4501 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     4561 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     4621 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     4681 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     4741 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc
     4801 atacccgttt ttttgggcta gaaataattt tgtttaactt taagaaggag atatac
//
