LOCUS       Exported                5936 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Encodes H2B-Dendra2-fusion protein under the CAG promoter. Linearize
            using Asc1 and Not1 for mouse embryo injection..
ACCESSION   .
VERSION     .
KEYWORDS    p55-H2B-Dendra2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5936)
  AUTHORS   Mohr MA, Argast P, Pantazis P
  TITLE     Labeling cellular structures in vivo using confined primed 
            conversion of photoconvertible fluorescent proteins.
  JOURNAL   Nat Protoc. 2016 Dec;11(12):2419-2431. doi: 10.1038/nprot.2016.134. 
            Epub 2016 Nov 3.
  PUBMED    27809312
REFERENCE   2  (bases 1 to 5936)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5936
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..666
                     /label=chicken beta-actin promoter
     intron          667..1675
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1599..1621
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1688..1706
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1723..2100
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             2119..2808
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     primer_bind     complement(2857..2874)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2863..3087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3159..3176)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3159..3175)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3168..3190)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3316..3771)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3453..3474)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3665..3684
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3798..3902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3903..4763
                     /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(4121..4140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4934..5522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5423..5442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5676..5693
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5810..5831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5846..5876
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5884..5900
                     /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     5889..5911
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5908..5924
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5908..5924
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cggattcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
      421 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
      481 gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag
      541 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
      601 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      661 cgggcgggag tcgctgcgtt gccttcgccc cgtgccccgc tccgcgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggctcg tttcttttct gtggctgcgt
      841 gaaagcctta aagggctccg ggagggccct ttgtgcgggg gggagcggct cggggggtgc
      901 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggcccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcgtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtgcggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctggcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcgggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccatct ccagcctcgg ggctgccgca
     1561 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1621 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagatcct
     1681 taattaataa tacgactcac tatagggggt cgacccgcca ccatgccaga gccagcgaag
     1741 tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc
     1801 ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg
     1861 aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat gaattcgttt
     1921 gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag
     1981 cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag
     2041 ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag
     2101 gatccaccgg tcgccaccat gaacaccccg ggaattaacc tgatcaagga ggacatgcgc
     2161 gtgaaggtgc acatggaggg caacgtgaac ggccacgcct tcgtgatcga gggcgagggc
     2221 aagggcaagc cctacgaggg cacccagacc gccaacctga ccgtgaagga gggcgccccc
     2281 ctgcccttca gctacgacat cctgaccacc gccgtgcact acggcaaccg ggtgttcacc
     2341 aagtaccccg aggacatccc cgactacttc aagcagagct tccccgaggg ctacagctgg
     2401 gagcgcacca tgaccttcga ggacaagggc atctgcacca tccgcagcga catcagcctg
     2461 gagggcgact gcttcttcca gaacgtgcgc ttcaagggca ccaacttccc ccccaacggc
     2521 cccgtgatgc agaagaagac cctgaagtgg gagcccagca ccgagaagct gcacgtgcgc
     2581 gacggcctgc tggtgggcaa catcaacatg gccctgctgc tggagggcgg cggccactac
     2641 ctgtgcgact tcaagaccac ctacaaggcc aagaaggtgg tgcagctgcc cgacgcccac
     2701 ttcgtggacc accgcatcga gatcctgggc aacgacagcg actacaacaa ggtgaagctg
     2761 tacgagcacg ccgtggcccg ctacagcccc ctgcccagcc aggtgtggtc cggcgacagc
     2821 ggcgtgtaca agtgagacgt cgagctcgct gatcagcctc gactgtgcct tctagttgcc
     2881 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2941 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3001 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3061 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     3121 gatccactag ttctagcctc gaggctagag cggccgccac tggccgtcgt tttacaacgt
     3181 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3241 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3301 ctgaatggcg aatggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3361 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3421 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3481 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3541 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3601 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3661 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3721 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3781 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3841 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3901 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3961 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4021 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4081 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4141 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4201 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4261 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4321 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4381 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4441 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4501 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4561 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4621 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4681 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4741 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4801 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4861 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4921 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4981 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5041 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5101 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5161 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5221 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5281 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5341 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5401 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5461 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5521 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5581 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5641 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5701 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5761 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     5821 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     5881 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgag gcgcgc
//
