LOCUS       sequence-61822-e        5089 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 5089)
  AUTHORS   Piatkevich KD, Malashkevich VN, Morozova KS, Nemkovich NA, Almo SC, 
            Verkhusha VV
  TITLE     Extended Stokes shift in fluorescent proteins: chromophore-protein 
            interactions in a near-infrared TagRFP675 variant.
  JOURNAL   Sci Rep. 2013;3:1847. doi: 10.1038/srep01847.
  PUBMED    23677204
REFERENCE   2  (bases 1 to 5089)
  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: "2013"; volume: "3:1847. doi"; pages: " 10.1038/srep01847"
COMMENT     Sequence Label: pH2B-TagRFP675-N1
FEATURES             Location/Qualifiers
     source          1..5089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             657..1034
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             1053..1754
                     /codon_start=1
                     /product="far-red monomeric fluorescent protein with an 
                     emission peak at 657 nm (Morozova et al., 2010)"
                     /label=TagRFP657
                     /note="derived from mKate"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQRIKV
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMMYPADGGLEGRNYMALK
                     LVGGGHLICSLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    1877..1998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1914..1933)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1968..1987
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2005..2460)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2142..2163)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2354..2373
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2487..2591
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2593..2950
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2593..2784
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2616..2636)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2801..2936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2985..3779
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2985..3779
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3039..3058)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3649..3668
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3967..3986)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4011..4058
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4387..4975
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4876..4895
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gccaccatgc
      661 cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg
      721 cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat tccatctatg
      781 tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag gccatgggca
      841 tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct tcccgcctgg
      901 cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc gtgcgcctgc
      961 tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc atcaccaagt
     1021 acaccagcgc taaggatcca ccggtcgcca ccatgagcga gctgattaag gagaacatgc
     1081 acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc acatccgagg
     1141 gcgaaggcaa gccctacgag ggcacccaga cccaaagaat caaggtggtc gagggcggcc
     1201 ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc aagaccttca
     1261 tcaaccacac ccagggcatc cccgacttct ggaagcagtc cttccctgag ggcttcacat
     1321 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     1381 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc ccatccaacg
     1441 gccctgtgat gcagaagaaa acactcggct gggaggccaa caccgagatg atgtaccccg
     1501 ctgacggcgg cctggaaggc agaaactaca tggccctgaa gctcgtgggc gggggccacc
     1561 tgatctgcag cttgaagacc acatacagat ccaagaaacc cgctaagaac ctcaagatgc
     1621 ccggcgtcta ctatgtggac cgcagactgg aaagaatcaa ggaggccgac aaagagacct
     1681 atgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc aaactggggc
     1741 acaagcttaa ttaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     1801 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     1861 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1921 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1981 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2041 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2101 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2161 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2221 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2281 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2341 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2401 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2461 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2521 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2581 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     2641 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2701 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2761 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2821 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2881 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2941 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3001 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3061 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3121 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3181 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3241 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3301 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3361 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3421 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3481 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     3541 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3601 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3661 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3721 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3781 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3841 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3901 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     3961 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4021 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4081 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4141 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4201 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4261 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4321 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4381 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4441 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4501 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     4561 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4621 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4681 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4741 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4801 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4861 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4921 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4981 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5041 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
