LOCUS       sequence-82259-e        4778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L-(+)-arabinose-inducible expression of link-CGFP negative control 
            for in vivo split GFP assembly assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4778)
  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 4778)
  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-link-CGFP
FEATURES             Location/Qualifiers
     source          1..4778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     224..244
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(338..356)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      352..399
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1001..1816)
                     /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(1001..1816)
                     /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     1724..1743
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1808..1827
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2277..2294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2411..2955)
                     /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(2491..2510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3535..4413)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4440..4724
                     /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     4647..4666
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4751..4773
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ccatggctaa tgcatgcaat gggacgtcgg gtggaagcgg taagaatgga atcaaagtga
       61 acttcaagac ccgccacaac attgaagatg gaagcgttca actagcagac cattatcaac
      121 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac
      181 aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg
      241 taacagctgc tgggattaca catggcatgg atgaactgta caactaagga tccggctgct
      301 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
      361 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
      421 ggatatccac aggacgggtg tggtcgccat gatggtgacc accatttgcc ccatatcacc
      481 gtcggtactg atcccgtcgt caataaaccg aaccgctaca ccctgagcat caaactcttt
      541 tatcagttgg atcatgtcgg cggtgtcgcg gccaagacgg tcgagcttct tcaccagaat
      601 gacatcacct tcctccacct tcatcctcag caaatccagc ccttcccgat ctgttgaact
      661 gccggatgcc ttgtcggtaa agatgcggtt agcttttacc cctgcatctt tgagcgctga
      721 ggtctgcctc gtgaagaagg tgttgctgac tcataccagg cctgaatcgc cccatcatcc
      781 agccagaaag tgagggagcc acggttgatg agagctttgt tgtaggtgga ccagttggtg
      841 attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg cgtgatctga
      901 tccttcaact cagcaaaagt tcgatttatt caacaaagcc gccgtcccgt caagtcagcg
      961 taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
     1021 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     1081 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     1141 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     1201 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     1261 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     1321 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     1381 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     1441 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     1501 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     1561 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     1621 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     1681 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     1741 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc
     1801 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     1861 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     1921 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     1981 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     2041 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     2101 cctggtatga gtcagcaaca ccttcttcac gaggcagacc tcagcgctag cggagtgtat
     2161 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     2221 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     2281 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     2341 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     2401 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     2461 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc ctggcggctc
     2521 cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc
     2581 gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg
     2641 actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
     2701 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
     2761 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
     2821 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
     2881 ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
     2941 ccaaaacgat ctcaagaaga tcatcttatt aaggggtctg acgctcagtg gaacgaaaac
     3001 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3061 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3121 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3181 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3241 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3301 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3361 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3421 gttgttgcca ttgctgcaga tcgatgcata atgtgcctgt caaatggacg aagcagggat
     3481 tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga
     3541 caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct
     3601 ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt
     3661 gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat
     3721 acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga
     3781 cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc
     3841 tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg
     3901 gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc
     3961 cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc
     4021 gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga
     4081 cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata
     4141 ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa
     4201 aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg
     4261 aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag
     4321 ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc
     4381 cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga
     4441 agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt
     4501 ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa
     4561 agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga
     4621 ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga
     4681 tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc
     4741 tagaaataat tttgtttaac tttaagaagg agatatac
//
