LOCUS       Exported                5992 bp ds-DNA     circular SYN 08-MAR-2023
DEFINITION  The fusion protein contains a SV40 NLS followed by mRFP, linker and 
            human PCNA..
ACCESSION   .
VERSION     .
KEYWORDS    pENmRFPCNAL2 pc1054
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5992)
  AUTHORS   Sporbert A, Domaing P, Leonhardt H, Cardoso MC
  TITLE     PCNA acts as a stationary loading platform for transiently 
            interacting Okazaki fragment maturation proteins.
  JOURNAL   Nucleic Acids Res. 2005 Jun 21;33(11):3521-8. doi: 
            10.1093/nar/gki665. Print 2005.
  PUBMED    15972794
REFERENCE   2  (bases 1 to 5992)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 8, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5992
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     7..26
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        49..352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        353..556
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     507..527
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..577
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     5'UTR           646..685
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             721..741
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             757..1431
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(887..905)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1155..1174
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1349..1372
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     intron          2305..2877
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3009..3028)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3074..3129
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3128..3147)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      3479..3614
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3533..3552)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3720..3737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3891..4479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3971..3990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4650..5510)
                     /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     5273..5292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5511..5615)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5683..5701
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5739..5761)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5861..5880
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgaaattaat acgactcact atagggagac cggaattcga gctcgccccg ttacataact
       61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      541 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
      601 ttttgacctc catagaagac accgggaccg atccagcctc cgggggatct tggtggcgtg
      661 aaactcccgc acctcttcgg ccagcgcctt gtagaagcgc gtatggcttc gtggggatcc
      721 ccgaagaaga agcgcaaagt actggtaccg gtcgacatgg cctcctccga ggacgtcatc
      781 aaggagttca tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag
      841 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      901 accaagggcg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt ccagtacggc
      961 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     1021 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     1081 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     1141 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
     1201 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agatgaggct gaagctgaag
     1261 gacggcggcc actacgacgc cgaggtcaag accacctaca tggccaagaa gcccgtgcag
     1321 ctgcccggcg cctacaagac cgacatcaag ctggacatca cctcccacaa cgaggactac
     1381 accatcgtgg aacagtacga gcgcgccgag ggccgccact ccaccggcgc cggcctgtac
     1441 aagggcgaag ggcaagggca agggcaaggg ccgggccgcg gctacgcgta tcgatccatg
     1501 ttcgaggcgc gcctggtcca gggctccatc ctcaagaagg tgttggaggc actcaaggac
     1561 ctcatcaacg aggcctgctg ggatattagc tccagcggtg taaacctgca gagcatggac
     1621 tcgtcccacg tctctttggt gcagctcacc ctgcggtctg agggcttcga cacctaccgc
     1681 tgcgaccgca acctggccat gggcgtgaac ctcaccagta tgtccaaaat actaaaatgc
     1741 gccggcaatg aagatatcat tacactaagg gccgaagata acgcggatac cttggcgcta
     1801 gtatttgaag caccaaacca ggagaaagtt tcagactatg aaatgaagtt gatggattta
     1861 gatgttgaac aacttggaat tccagaacag gagtacagct gtgtagtaaa gatgccttct
     1921 ggtgaatttg cacgtatatg ccgagatctc agccatattg gagatgctgt tgtaatttcc
     1981 tgtgcaaaag acggagtgaa attttctgca agtggagaac ttggaaatgg aaacattaaa
     2041 ttgtcacaga caagtaatgt cgataaagag gaggaagctg ttaccataga gatgaatgaa
     2101 ccagttcaac taacttttgc actgaggtac ctgaacttct ttacaaaagc cactccactc
     2161 tcttcaacgg tgacactcag tatgtctgca gatgtacccc ttgttgtaga gtataaaatt
     2221 gcggatatgg gacacttaaa atactacttg gctcccaaga tcgaggatga agaaggatct
     2281 tagtctagag ctgagaactt cagggtgagt ttggggaccc ttgattgttc tttctttttc
     2341 gctattgtaa aattcatgtt atatggaggg ggcaaagttt tcagggtgtt gtttagaatg
     2401 ggaagatgtc ccttgtatca ccatggaccc tcatgataat tttgtttctt tcactttcta
     2461 ctctgttgac aaccattgtc tcctcttatt ttcttttcat tttctgtaac tttttcgtta
     2521 aactttagct tgcatttgta acgaattttt aaattcactt ttgtttattt gtcagattgt
     2581 aagtactttc tctaatcact tttttttcaa ggcaatcagg gtatattata ttgtacttca
     2641 gcacagtttt agagaacaat tgttataatt aaatgataag gtagaatatt tctgcatata
     2701 aattctggct ggcgtggaaa tattcttatt ggtagaaaca actacaccct ggtcatcatc
     2761 ctgcctttct ctttatggtt acaatgatat acactgtttg agatgaggat aaaatactct
     2821 gagtccaaac cgggcccctc tgctaaccat gttcatgcct tcttctcttt cctacagctc
     2881 ctgggcaacg tgctggttgt tgtgctgtct catcattttg gcaaagaatt cactcctcag
     2941 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     3001 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     3061 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     3121 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     3181 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     3241 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     3301 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     3361 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     3421 ccagtcatag ctgtccctct tctcttatgg agatcccgga gctttttgca aaagcctagg
     3481 cctccaaaaa agcctcttca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     3541 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     3601 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgctgcat
     3661 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3721 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3781 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3841 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3901 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3961 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4021 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4081 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4141 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4201 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4261 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4321 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4381 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4441 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4501 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4561 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4621 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4681 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4741 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4801 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4861 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4921 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4981 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5041 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5101 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5161 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5221 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5281 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5341 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5401 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5461 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5521 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5581 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5641 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5701 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5761 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5821 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     5881 ctgagagtgc accattcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     5941 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtctggctt at
//
