LOCUS       sequence-332748-        6346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-fragment of the DiB2‐split protein fused with blue fluorescent 
            protein TagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6346)
  AUTHORS   Bozhanova NG, Gavrikov AS, Mishin AS, Meiler J
  TITLE     DiB-splits: nature-guided design of a novel fluorescent labeling 
            split system.
  JOURNAL   Sci Rep. 2020 Jul 6;10(1):11049. doi: 10.1038/s41598-020-67095-2.
  PUBMED    32632329
REFERENCE   2  (bases 1 to 6346)
  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: "2020-07-6"; volume: "10(1):11049. doi"; pages: " 
            10.1038/s41598-020-67095-2"
COMMENT     Sequence Label: TagBFP-DiB2-splitN1–125
FEATURES             Location/Qualifiers
     source          1..6346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(288..1079,1080..1148))
                     /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     911..930
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1149..1253)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1340..1971
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    2063..2087
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     enhancer        2196..2499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2500..2703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2654..2674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2748..3446
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    3905..4026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3942..3961)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3996..4015
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    4211..4235
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4391..5539)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(join(5875..6346,1..117))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5955..5974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
       61 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      121 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      181 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      241 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      301 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      361 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      421 ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca gccagccgga
      481 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      541 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      601 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      661 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      721 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
      781 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
      841 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
      901 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
      961 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1021 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1081 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1141 atactcatac tcttcctttt tcaatattat tgaagcattt atcaaggtta ttgtctcatg
     1201 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacgaat
     1261 tggccagcgc tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg
     1321 gggatgggag gcccgcgtta gcgggccggg agggttcgag aagggggggc accccccttc
     1381 ggcgtgcgcg gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt tataaatatt
     1441 ggtttaaaag caggttaaaa gacaggttag cggtggccga aaaacgggcg gaaacccttg
     1501 caaatgctgg attttctgcc tgtggacagc ccctcaaatg tcaataggtg cgcccctcat
     1561 ctgtcagcac tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg
     1621 cccctcaagt gtcaataccg cagggcactt atccccaggc ttgtccacat catctgtggg
     1681 aaactcgcgt aaaatcaggc gttttcgccg atttgcgagg ctggccagct ccacgtcgcc
     1741 ggccgaaatc gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa
     1801 gtgtcaacgt ccgcccctca tctgtcagtg agggccaagt tttccgcgag gtatccacaa
     1861 cgccggcggc cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg
     1921 ccatagacgg ccgccagccc agcggcgagg gcaaccagcc cggtgagcgt cgcaaaggag
     1981 atcctgatct gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
     2041 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta
     2101 gacaacttaa taacacattg cggacgtttt taatgtactg gggtggatgc agtgggcccc
     2161 actctgtgaa gacaatgccg aattcggatc cggagcgtta cataacttac ggtaaatggc
     2221 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2281 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2341 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2401 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2461 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2521 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2581 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     2641 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     2701 gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg agcgagctga
     2761 ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca
     2821 agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg
     2881 tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg
     2941 gcagcaagac cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc
     3001 ctgagggctt cacatgggag agagtcacca catacgagga cgggggcgtg ctgaccgcta
     3061 cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga
     3121 acttcacatc caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg
     3181 agacgctgta ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg
     3241 tgggcgggag ccatctgatc gcaaacatca agaccacata tagatccaag aaacccgcta
     3301 agaacctcaa gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg
     3361 ccaacaacga aacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc
     3421 ctagcaaact ggggcacaag cttaatgatc ccccggtcgc cacaatggcc agtagcccga
     3481 ctcctccgcg aggagttact gtggttaaca attttgactg caaacgctat ctcgggacgt
     3541 ggtatgagat agcccgattc gatcacaggt tcgaacgcgg attggagaaa gttaccgcaa
     3601 cctatagttt gcgggacgac gggggcttga acgtaatcaa caaagggtat aatccagaca
     3661 ggggcatgtg gcagcagagc gaaggtaaag cgtacttcac gggtgccccc acgagggcgg
     3721 cattgaaggt cagcttcttc ggaccttttt atggtggcta taacgtaata gcattggaca
     3781 gggagtattc aggttgatga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3841 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3901 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3961 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4021 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4081 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4141 aatagaccga gatagggttg agtgtcgctg caattgtctt ctgcacgaag tggtttaaac
     4201 tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga
     4261 ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat
     4321 gccaaccaca gggttcccca gatcaggcgc tggctgctga acccccagcc ggaactgacc
     4381 ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg tgttccacca
     4441 ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc
     4501 gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg tacggctccc
     4561 ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg cggtacttct
     4621 cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc tcgtcgatca
     4681 ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg
     4741 acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc
     4801 gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag cccaggtcct
     4861 ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact
     4921 ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg
     4981 tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga
     5041 ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc atcgctcgca
     5101 tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg atctgctgct
     5161 tcgtgtgttt cagcaacgcg gcctgcttgg cctcgctgac ctgttttgcc aggtcctcgc
     5221 cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg
     5281 ccaaacctgc cgcctcctgt tcaagacgac gcgaacgctc cacggcggcc gatggcgcgg
     5341 gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta gcttgctgga
     5401 ccatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg cggcttgcga
     5461 tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc
     5521 ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccgactcac gccggggcaa
     5581 tgtgccctta ttcctgattt gacccgcctg gtgccttggt gtccagataa tccaccttat
     5641 cggcaatgaa gtcggtcccg tagaccgtct ggccgtcctt ctcgtacttg gtattccgaa
     5701 tcttgccctg cacgaatacc agcgacccct tgcccaaata cttgccgtgg gcctcggcct
     5761 gagagccaaa acacttgatg cggaagaagt cggtgcgctc ctgcttgtcg ccggcatcgt
     5821 tgcgccacat ctaggatctg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaag
//
