LOCUS       sequence-470397-        6262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human PCNA miRFP670  tagged in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6262)
  AUTHORS   Rausch C, Zhang P, Casas-Delucchi CS, Daiss JL, Engel C, Coster G, 
            Hastert FD, Weber P, Cardoso MC
  TITLE     Cytosine base modifications regulate DNA duplex stability and 
            metabolism.
  JOURNAL   Nucleic Acids Res. 2021 Dec 16;49(22):12870-12894. doi: 
            10.1093/nar/gkab509.
  PUBMED    34133727
REFERENCE   2  (bases 1 to 6262)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2021-12-16- 16"; volume: "49"; issue: 
            "22"; pages: "12870-12894"
COMMENT     Sequence Label: pmiRFP670-PCNAL2 (pc3385)
FEATURES             Location/Qualifiers
     source          1..6262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      866..1001
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1333..1352
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1351..1406
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1452..1471
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          1603..2175
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             complement(3046..3990)
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     regulatory      3987..3996
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4009..4029)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     5'UTR           4065..4104
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     primer_bind     complement(4173..4197)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4194..4397)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4223..4243)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4398..4701
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4724..4743)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4725..4743)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4862..4881)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4981..5003
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5041..5059)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5127..5231
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5232..5300,5301..6092)
                     /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(5450..5469)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcaatt agtcagcaac
      841 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      901 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
      961 tgagctattc cagaagtagt gaagaggctt ttttggaggc ctaggctttt gcaaaaagct
     1021 ccgggatctc cataagagaa gagggacagc tatgactggg agtagtcagg agaggaggaa
     1081 aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac
     1141 aaaacaaaat ataaaaaaaa tctaacctca agtcaaggct tttctatgga ataaggaatg
     1201 gacagcaggg ggctgtttca tatactgatg acctctttat agccaacctt tgttcatggc
     1261 agccagcata tgggcatatg ttgccaaact ctaaaccaaa tactcattct gatgttttaa
     1321 atgatttgcc ctcccatatg tccttccgag tgagagacac aaaaaattcc aacacactat
     1381 tgcaatgaaa ataaatttcc tttattagcc agaagtcaga tgctcaaggg gcttcatgat
     1441 gtccccataa tttttggcag agggaaaaag atctcagtgg tatttgtgag ccagggcatt
     1501 ggccacacca gccaccacct tctgataggc agcctgcacc tgaggagtga attctttgcc
     1561 aaaatgatga gacagcacaa caaccagcac gttgcccagg agctgtagga aagagaagaa
     1621 ggcatgaaca tggttagcag aggggcccgg tttggactca gagtatttta tcctcatctc
     1681 aaacagtgta tatcattgta accataaaga gaaaggcagg atgatgacca gggtgtagtt
     1741 gtttctacca ataagaatat ttccacgcca gccagaattt atatgcagaa atattctacc
     1801 ttatcattta attataacaa ttgttctcta aaactgtgct gaagtacaat ataatatacc
     1861 ctgattgcct tgaaaaaaaa gtgattagag aaagtactta caatctgaca aataaacaaa
     1921 agtgaattta aaaattcgtt acaaatgcaa gctaaagttt aacgaaaaag ttacagaaaa
     1981 tgaaaagaaa ataagaggag acaatggttg tcaacagagt agaaagtgaa agaaacaaaa
     2041 ttatcatgag ggtccatggt gatacaaggg acatcttccc attctaaaca acaccctgaa
     2101 aactttgccc cctccatata acatgaattt tacaatagcg aaaaagaaag aacaatcaag
     2161 ggtccccaaa ctcaccctga agttctcagc tctagactaa gatccttctt catcctcgat
     2221 cttgggagcc aagtagtatt ttaagtgtcc catatccgca attttatact ctacaacaag
     2281 gggtacatct gcagacatac tgagtgtcac cgttgaagag agtggagtgg cttttgtaaa
     2341 gaagttcagg tacctcagtg caaaagttag ttgaactggt tcattcatct ctatggtaac
     2401 agcttcctcc tctttatcga cattacttgt ctgtgacaat ttaatgtttc catttccaag
     2461 ttctccactt gcagaaaatt tcactccgtc ttttgcacag gaaattacaa cagcatctcc
     2521 aatatggctg agatctcggc atatacgtgc aaattcacca gaaggcatct ttactacaca
     2581 gctgtactcc tgttctggaa ttccaagttg ttcaacatct aaatccatca acttcatttc
     2641 atagtctgaa actttctcct ggtttggtgc ttcaaatact agcgccaagg tatccgcgtt
     2701 atcttcggcc cttagtgtaa tgatatcttc attgccggcg cattttagta ttttggacat
     2761 actggtgagg ttcacgccca tggccaggtt gcggtcgcag cggtaggtgt cgaagccctc
     2821 agaccgcagg gtgagctgca ccaaagagac gtgggacgag tccatgctct gcaggtttac
     2881 accgctggag ctaatatccc agcaggcctc gttgatgagg tccttgagtg cctccaacac
     2941 cttcttgagg atggagccct ggaccaggcg cgcctcgaac atggatcgat acgcgtagcc
     3001 gcggcccggc ccttgccctt gcccttgccc ttcgcccttg tacaggctct caagcgcggt
     3061 gatccgcgtc gcgatccttt cggcgagccg tttgcagatc gcccgcagct cgaaacggat
     3121 gaagcgcggc agataatggt gacagacaac caggccccac agcttgccgc cgaccaccag
     3181 cgacaccgcc agggtggcgc gcacgcccat gtccttcagg aactgcagat ggcacggcga
     3241 catcgagcgc aggaagcagc ccgacatgtc gagatcgcgc ccggtcagcg gcgacagccg
     3301 cggctccagc ggcaccggct gataggtgac gtcgaccagc acgcggacgc gctgccgcac
     3361 gtacagctgc cgcgccatct gcgggacagt cgacgacgga tagcggttgc cgaaatagga
     3421 ttcgagccca ggcacatggc actcggagaa taccaggccg tggccttgct catcgaaacg
     3481 atacaccatc acccggtcgt agccggtgca ctgctgaaac agcagcacgg tgtcatcgca
     3541 cagcgcgcgc agtgaacccg ccgtgcggat ccgctccagc gccggcgcca gcgtgcctga
     3601 cagatcgatc gacgggccgg cacgttcgag ttcgatgatc agcccgcctt ccggaggccg
     3661 atgcatcaga ccgcagtact ccgtagaggg attgccgatc cggcagcgca ccgcgaccgg
     3721 catgccttcg gcggtgggat cgagatgcgg caggatcttg atcaacagat cgccgtcgat
     3781 ctcggcgagc ggaacgccga gtacgcttcc gagattcaga aattccgcgg cgttggcgct
     3841 ggcctggatg acgcgatgat catgttcgct gacgaccaga agcgcgccat gcggctggat
     3901 cgagccggcg aggtggatct cttcatgttc gcaattcgaa tgagaggcgg tcccgaatgc
     3961 ggggctgcca gaggcatgac ctgctaccat ggtggcgacc ggtaccagta ctttgcgctt
     4021 cttcttcggg gatccccacg aagccatacg cgcttctaca aggcgctggc cgaagaggtg
     4081 cgggagtttc acgccaccaa gatcccccgg aggctggatc ggtcccggtg tcttctatgg
     4141 aggtcaaaac agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg
     4201 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     4261 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     4321 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     4381 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     4441 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     4501 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     4561 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     4621 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     4681 tttaccgtaa gttatgtaac ggggcgagct cgaattccgg tctccctata gtgagtcgta
     4741 ttaatttcga taagccagac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     4801 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgaatgg tgcactctca
     4861 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4921 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4981 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5041 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5101 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5161 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5221 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5281 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5341 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5401 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5461 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5521 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5581 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5641 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5701 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5761 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5821 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5881 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5941 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6001 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6061 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6121 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6181 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6241 tagaaaagat caaaggatct tc
//
