LOCUS       sequence-222824-        4457 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 4457)
  AUTHORS   Kodama Y, Hu CD
  TITLE     An improved bimolecular fluorescence complementation assay with a 
            high signal-to-noise ratio.
  JOURNAL   Biotechniques. 2010 Nov . 49(5):793-805.
  PUBMED    21091444
REFERENCE   2  (bases 1 to 4457)
  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: 
            "Biotechniques. 2010 Nov . 49(5):793-805."
COMMENT     Sequence Label: pBiFC-bJunVN155(I152L)
FEATURES             Location/Qualifiers
     source          1..4457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(289..291,292..294,295..753)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYLTA"
     primer_bind     complement(334..355)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(595..614)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    768..902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(792..811)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     846..865
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1013..1029)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1013..1029)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1026..1048)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1037..1053
                     /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(1061..1067,1068..1085,1086..1091))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1106..1127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1244..1261)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1415..2003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1495..1514)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2174..2965,2966..3034))
                     /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     2797..2816
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3035..3139)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3207..3225
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3263..3285)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3385..3404
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3598..3620
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3612..3629
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3613..3629
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3656..3959
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3960..4163
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4114..4134
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4160..4184
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4301..4397
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
ORIGIN
        1 atggcatcaa tgcagaagct gatctcagag gaggacctgc ttatggccat ggaggcccga
       61 attcggaagg cggagaggaa gcgcatgagg aaccgcatcg ctgcctccaa gtgccggaaa
      121 aggaagctgg agcggatcgc ccggctagag gaaaaagtga aaaccttgaa agcgcaaaac
      181 tccgagctgg cgtccacggc caacatgctc agggaacagg tggcacagct taaacagaaa
      241 gtcatgaacc acggtaccgg aggtggcggg agcggaggtg gcgggagtat ggtgagcaag
      301 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      361 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      421 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      481 ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      541 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      601 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      661 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      721 aactacaaca gccacaacgt ctatctcacc gcctgagcgg ccgcggggat ccagacatga
      781 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      841 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      901 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      961 tttcggatcc tctagagtcg atctgcaggc atgctagctt ggcgtaatca tggtcatagc
     1021 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     1081 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     1141 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     1201 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     1261 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1321 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1381 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1441 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1501 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1561 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1621 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1681 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1741 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1801 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1861 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1921 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1981 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2041 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2101 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2161 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2221 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2281 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2341 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2401 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2461 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2521 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2581 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2641 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2701 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2761 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2821 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2881 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2941 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3001 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3061 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3121 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     3181 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     3241 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3301 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3361 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     3421 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     3481 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3541 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     3601 agtcacgacg ttgtaaaacg acggccagtg agttcgagct tgcatgcctg caggtcgtta
     3661 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     3721 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     3781 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     3841 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     3901 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     3961 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4021 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4081 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     4141 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     4201 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg actctagagg
     4261 atccggtact agaggaactg aaaaaccaga aagttaactg gtaagtttag tctttttgtc
     4321 ttttatttca ggtcccggat ccggtggtgg tgcaaatcaa agaactgctc ctcagtggat
     4381 gttgccttta cttctaggcc tgtacggaag tgttacttct gctctaaaag ctgcggaatt
     4441 gtacccgcgg gcccacc
//
