LOCUS       sequence-10109-e        4515 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 4515)
  AUTHORS   Shyu YJ, Hiatt SM, Duren HM, Ellis RE, Kerppola TK, Hu CD
  TITLE     Visualization of protein interactions in living Caenorhabditis 
            elegans using bimolecular fluorescence complementation analysis.
  JOURNAL   Nat Protoc. 2008  . 3(4):588-96.
  PUBMED    18388940
REFERENCE   2  (bases 1 to 4515)
  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: "Nat Protoc.
            2008  . 3(4):588-96."
COMMENT     Sequence Label: pCE-BiFC-VN173
FEATURES             Location/Qualifiers
     source          1..4515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(136..138,139..141,142..654)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(181..202)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(442..461)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1522..1544)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1644..1663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1741..1759)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1827..1931
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1932..2000,2001..2792)
                     /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     complement(2150..2169)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2963..3551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3452..3471
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3705..3722
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3839..3860
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3875..3880,3881..3898,3899..3905)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3913..3929
                     /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)."
     primer_bind     3918..3940
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3937..3953
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3937..3953
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atggcatcaa tgcagaagct gatctcagag gaggacctgc ttatggccat ggaggcccga
       61 attcggtcga ccgagatctc tcgaggtacc ggtggcggag gttctggtgg cggaggttct
      121 ggtggcggag gttctatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      181 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      241 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
      301 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
      361 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      421 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      481 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      541 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
      601 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgagtaagcg
      661 gccgcaggat ccagacatga taagatacga tatctgagct ccgcatcggc cgctgtcatc
      721 agatcgccat ctcgcgcccg tgcctctgac ttctaagtcc aattactctt caacatccct
      781 acatgctctt tctccctgtg ctcccacccc ctatttttgt tattatcaaa aaaacttctt
      841 cttaatttct ttgtttttta gcttctttta agtcacctct aacaatgaaa ttgtgtagat
      901 tcaaaaatag aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc tccccccatt
      961 aataataatt ctatcccaaa atctacacaa tgttctgtgt acacttctta tgtttttttt
     1021 acttctgata aatttttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca
     1081 tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat
     1141 gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa
     1201 gtgaaagttt atgaaattaa ttttcctgct tttgcttttt gggggtttcc cctattgttt
     1261 gtcaagagtt tcgaggacgg cgtttttctt gctaaaatca caagtattga tgagcacgat
     1321 gcaagaaaga tcggaagaag gtttgggttt gaggctcagt ggaaggtgag tagaagttga
     1381 taatttgaaa gtggagtagt gtctatgggg tttttgcctt aaatgacaga atacattccc
     1441 aatataccaa acataactgt ttcctactag tcggccgtac gggccctttc gtctcgcgcg
     1501 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     1561 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     1621 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     1681 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg
     1741 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     1801 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1861 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1921 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1981 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2041 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2101 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2161 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2221 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2281 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2341 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2401 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2461 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2521 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2581 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2641 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2701 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2761 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2821 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2881 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2941 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3001 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3061 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3121 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3181 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3241 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3301 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag
     3361 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3421 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3481 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3541 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3601 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3661 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3721 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3781 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3841 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     3901 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     3961 acgccaagct tgcatgcctg caggtcgact ctagaggatc accaaaaacg gaacgttgag
     4021 ctggacggaa atagtggtaa agtgacatga ttatagtttg aagatttcta atttcacaat
     4081 tagagcaaat gttgttcggt atttattttc aacggtattt atactatttt ccaccttttt
     4141 ctagaacatt cgagctgctt gttgcaaaag gagggcgact cacattcggt acatggaaaa
     4201 gtagtgtaca caataaagag acccagatac attttccgtc tgcgtctctt tgcacccacc
     4261 gggagtattt tcaaacgaat gcatctagga ccttctagaa cattctgtaa ggctgcagaa
     4321 tgcgggtata taaggaaagc gggctcagag gaagccaaca cgctttgttc tagtgcatct
     4381 aaaaaacttc gaaaatcctc atcgggatcc ccgggattgg ccaaaggacc caaaggtatg
     4441 tttcgaatga tactaacata acatagaaca ttttcaggag gacccttggc tagcaattgt
     4501 acccgcgggc ccacc
//
