LOCUS       sequence-332747-        6025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-fragment of the DiB2‐split protein fused with histone H2B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6025)
  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 6025)
  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: H2B-DiB2-splitN1–125
FEATURES             Location/Qualifiers
     source          1..6025
                     /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..3125
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     polyA_signal    3584..3705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3621..3640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3675..3694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3890..3914
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4070..5218)
                     /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(5554..6025,1..117))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5634..5653)
                     /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 ccagagccag
     2761 cgaagtctgc tcccgccccg aaaaagggct ccaagaaggc ggtgactaag gcgcagaaga
     2821 aaggcggcaa gaagcgcaag cgcagccgca aggagagcta ttccatctat gtgtacaagg
     2881 ttctgaagca ggtccaccct gacaccggca tttcgtccaa ggccatgggc atcatgaact
     2941 cgtttgtgaa cgacattttc gagagaatcg caggtgaggc ttcccgcctg gcgcattaca
     3001 acaagcgctc gaccatcacc tccagggaga tccagacggc cgtgcgcctg ctgctgcctg
     3061 gggagttggc caagcacgcc gtgtccgagg gtactaaggc catcaccaag tacaccagcg
     3121 ctaaagatcc cccggtcgcc acaatggcca gtagcccgac tcctccgcga ggagttactg
     3181 tggttaacaa ttttgactgc aaacgctatc tcgggacgtg gtatgagata gcccgattcg
     3241 atcacaggtt cgaacgcgga ttggagaaag ttaccgcaac ctatagtttg cgggacgacg
     3301 ggggcttgaa cgtaatcaac aaagggtata atccagacag gggcatgtgg cagcagagcg
     3361 aaggtaaagc gtacttcacg ggtgccccca cgagggcggc attgaaggtc agcttcttcg
     3421 gaccttttta tggtggctat aacgtaatag cattggacag ggagtattca ggttgatgat
     3481 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3541 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3601 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3661 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     3721 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3781 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3841 gtgtcgctgc aattgtcttc tgcacgaagt ggtttaaact atcagtgttt gacaggatat
     3901 attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg
     3961 cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttccccag
     4021 atcaggcgct ggctgctgaa cccccagccg gaactgaccc cacaaggccc tagcgtttgc
     4081 aatgcaccag gtcatcattg acccaggcgt gttccaccag gccgctgcct cgcaactctt
     4141 cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca
     4201 tgaggcggaa ggtttccagc ttgagcgggt acggctcccg gtgcgagctg aaatagtcga
     4261 acatccgtcg ggccgtcggc gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt
     4321 ggtcgccagc aaacagcacg acgatttcct cgtcgatcag gacctggcaa cgggacgttt
     4381 tcttgccacg gtccaggacg cggaagcggt gcagcagcga caccgattcc aggtgcccaa
     4441 cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct
     4501 tcgtgtaata ccggccattg atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga
     4561 aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc caacacctgc tgccacacca
     4621 gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg tccttgttga
     4681 cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga
     4741 acagggcaga gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa
     4801 tatcgaacaa ggaaagctgc atttccttga tctgctgctt cgtgtgtttc agcaacgcgg
     4861 cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg
     4921 tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt
     4981 caagacgacg cgaacgctcc acggcggccg atggcgcggg cagggcaggg ggagccagtt
     5041 gcacgctgtc gcgctcgatc ttggccgtag cttgctggac catcgagccg acggactgga
     5101 aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg
     5161 aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc cgattcattc
     5221 accctccttg cgggattgcc ccgactcacg ccggggcaat gtgcccttat tcctgatttg
     5281 acccgcctgg tgccttggtg tccagataat ccaccttatc ggcaatgaag tcggtcccgt
     5341 agaccgtctg gccgtccttc tcgtacttgg tattccgaat cttgccctgc acgaatacca
     5401 gcgacccctt gcccaaatac ttgccgtggg cctcggcctg agagccaaaa cacttgatgc
     5461 ggaagaagtc ggtgcgctcc tgcttgtcgc cggcatcgtt gcgccacatc taggatctgc
     5521 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5581 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5641 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5701 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5761 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5821 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5881 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5941 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6001 atttggtatc tgcgctctgc tgaag
//
