LOCUS       sequence-10104-e        5420 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 5420)
  AUTHORS   Shyu YJ, Liu H, Deng X, Hu CD
  TITLE     Identification of new fluorescent protein fragments for bimolecular 
            fluorescence complementation analysis under physiological 
            conditions.
  JOURNAL   Biotechniques. 2006 Jan . 40(1):61-6.
  PUBMED    16454041
REFERENCE   2  (bases 1 to 5420)
  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. 2006 Jan . 40(1):61-6."
COMMENT     Sequence Label: pBiFC-bJunVN173
FEATURES             Location/Qualifiers
     source          1..5420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             935..958
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1226..1235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1232..1234,1235..1237,1238..1750)
                     /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(1277..1298)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1538..1557)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    1761..2383
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1960..1979)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2407..2427)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2412..2741
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2592..2727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2779..2798)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2780..2798)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2812..2828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2812..2828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2825..2847)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2836..2852
                     /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(2860..2866,2867..2884,2885..2890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2905..2926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3043..3060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3214..3802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3294..3313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3973..4764,4765..4833))
                     /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     4596..4615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4834..4938)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4965..5393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5052..5071)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5262..5283
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
      961 tgcggccgcg aattcaaagg cggagaggaa gcgcatgaga aaccgcatcg ctgcctccaa
     1021 gtgccggaaa aggaagctgg agcggatcgc ccggctagag gaaaaagtga aaaccttgaa
     1081 agcgcaaaac tccgagctgg cgtccacggc caacatgctc agggaacagg tggcacagct
     1141 taaacagaaa gtcatgaacc acgttaacag tgggtgccaa ctcatgctaa cgcagcagtt
     1201 gcaaacgttt tctagaagat ccatcgccac catggtgagc aagggcgagg agctgttcac
     1261 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1321 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     1381 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcctgca
     1441 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1501 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1561 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1621 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1681 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     1741 caacatcgag tagggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     1801 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     1861 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     1921 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     1981 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     2041 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     2101 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     2161 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     2221 ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac
     2281 agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg
     2341 tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt
     2401 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2461 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2521 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2581 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2641 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2701 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga
     2761 aaaaccagaa agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct
     2821 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2881 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2941 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3001 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3061 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3121 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3841 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3901 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3961 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4021 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4081 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4141 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4201 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4261 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4321 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4381 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4441 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4501 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4561 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4621 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4681 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4741 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4801 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4861 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4921 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     4981 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5041 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5101 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5161 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5221 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5281 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5341 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5401 aatattaacg cttacaattt
//
