LOCUS       sequence-359601-        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tripartite split-GFP mammalian. Express leucine zipper with GFP10 
            and GFP11 (self-assembing with GFP1-9).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5296)
  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 5296)
  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-GFP11
FEATURES             Location/Qualifiers
     source          1..5296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(98..118)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        103..432
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      283..418
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     345..364
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        480..527
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             546..920
                     /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    1050..1171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1087..1106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1141..1160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1220..1236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1220..1236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1233..1255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1244..1260
                     /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(1268..1274,1275..1292,1293..1298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1313..1334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1451..1468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1622..2210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1702..1721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2381..3172,3173..3241))
                     /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     3004..3023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3242..3346)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3421..3440)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3612..3991
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3992..4195
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4146..4166
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4240..4259
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4240..4258
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4278..4287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4470..4526
                     /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(4681..4698)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4687..4911
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(4957..5296,1..89)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5044..5063)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5254..5275
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
       61 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
      121 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
      181 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
      241 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
      301 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
      361 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
      421 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
      481 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
      541 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
      601 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
      661 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
      721 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
      781 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
      841 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
      901 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
      961 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     1021 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     1081 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1141 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     1201 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1261 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1321 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1381 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1441 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1501 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1561 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1621 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1741 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1801 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1861 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1921 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1981 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2041 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2101 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2161 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2221 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2281 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2341 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2401 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2461 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2521 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2581 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2641 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2701 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2761 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2821 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2881 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2941 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3001 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3061 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3121 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3181 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3241 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3301 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3361 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     3421 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     3481 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     3541 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     3601 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     3661 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     3721 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     3781 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     3841 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3901 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3961 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     4021 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     4081 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     4141 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     4201 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     4261 gacccaagct ggctagcgcc accatgggcg acctgcccga cgaccactac ctgtccaccc
     4321 agaccatcct gagcaaggag ctgaacatcg atggtggcgg tggctctgga ggtggtgggt
     4381 ccgcggccgc gggaggtatg aacactgaag ccgccaggcg ttctcgtgcg agaaagttgc
     4441 aaagaatgaa acaacttgaa gacaaggttg aagaattgct ttcgaaaaat tatcacttgg
     4501 aaaatgaggt tgccagatta aagaaattag ttggcgaacg catcgatggt ggcggtggct
     4561 ctggaggtgg tgggtcctcc ggagagaagc gcgaccacat ggtgctgctg gagtacgtga
     4621 ccgccgccgg catcaccgac gcctcctaat ctagagggcc cgtttaaacc cgctgatcag
     4681 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4741 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4801 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4861 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4921 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4981 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5041 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5101 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5161 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5221 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5281 cactcaaccc tatctc
//
