LOCUS       sequence-359553-        5182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tripartite split-GFP mammalian. Express leucine zipper with GFP10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5182)
  AUTHORS   Cabantous S, Nguyen HB, Pedelacq JD, Koraichi F, Chaudhary A, 
            Ganguly K, Lockard MA, Favre G, Terwilliger TC, Waldo GS
  TITLE     A new protein-protein interaction sensor based on tripartite 
            split-GFP association.
  JOURNAL   Sci Rep. 2013 Oct 4;3:2854. doi: 10.1038/srep02854.
  PUBMED    24092409
REFERENCE   2  (bases 1 to 5182)
  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: "Sci Rep."; 
            date: "2013-10-4"; volume: "3:2854. doi"; pages: " 
            10.1038/srep02854"
COMMENT     Sequence Label: pcDNA GFP10-ZIP
FEATURES             Location/Qualifiers
     source          1..5182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(41..60)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        232..611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        612..815
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     766..786
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     860..879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        860..878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      898..907
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1081..1137
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     primer_bind     complement(1187..1204)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1193..1417
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1463..1891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1550..1569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1760..1781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1900..1920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1905..2234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2085..2220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2147..2166
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2282..2329
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2348..2722
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2852..2973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2889..2908)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2943..2962
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3022..3038)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3022..3038)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3035..3057)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3046..3062
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3070..3076,3077..3094,3095..3100))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3115..3136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3253..3270)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3424..4012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3504..3523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4183..4974,4975..5043))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4806..4825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5044..5148)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
       61 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
      121 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
      181 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      241 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      301 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      361 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      421 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      481 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      541 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      601 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      661 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      721 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      781 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      841 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagcgcc
      901 accatgggcg acctgcccga cgaccactac ctgtccaccc agaccatcct gagcaaggac
      961 ctgaacatcg atggtggcgg tggctctgga ggtggtgggt cctccggaat gaacactgaa
     1021 gccgccaggc gttctcgtgc gagaaagttg caaagaatga aacaacttga agacaaggtt
     1081 gaagaattgc tttcgaaaaa ttatcacttg gaaaatgagg ttgccagatt aaagaaatta
     1141 gttggcgaac gctaatctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     1201 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1261 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1321 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1381 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     1441 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1501 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1561 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1621 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1681 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1741 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1801 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1861 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     1921 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     1981 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2041 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2161 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2221 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     2281 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     2341 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     2401 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     2461 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     2521 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     2581 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     2641 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     2701 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
     2761 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     2821 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     2881 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2941 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     3001 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3061 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3121 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3181 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3241 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3301 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3361 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3421 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3481 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3541 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3601 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3661 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3721 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3781 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3841 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3901 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3961 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4021 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4081 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4141 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4201 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4261 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4321 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4381 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4441 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4501 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4561 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4621 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4681 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4741 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4801 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4861 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4921 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4981 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5041 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5101 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5161 aaaagtgcca cctgacgtcg ac
//
