LOCUS       sequence-228927-        5580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus/Replication Origins, Excitation: 400 / 504, 
            Emission: 515 / 517.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5580)
  TITLE     Davidson Photoactivatable Fluorescent Proteins
REFERENCE   2  (bases 1 to 5580)
  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"
COMMENT     Sequence Label: PA-mCherry1-PCNA-19-SV40NLS-4
FEATURES             Location/Qualifiers
     source          1..5580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      535..544
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             547..567
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             571..591
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             595..615
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      622..631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             628..1335
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein (Subach et al., 2009)"
                     /label=PAmCherry1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(773..791)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2308..2429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2345..2364)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2399..2418
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2436..2891)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2573..2594)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2785..2804
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2918..3022
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3024..3381
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3024..3215
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3047..3067)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3232..3367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3416..4210
                     /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             3416..4210
                     /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(3470..3489)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4080..4099
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4398..4417)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4442..4489
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4818..5406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5307..5326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccacc
      541 atggatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt agatccaaaa
      601 aagaagagaa aggtaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc
      661 atcattaagg agttcatgcg cttcaaggtg cacatggagg ggtccgtgaa cggccacgtg
      721 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      781 aaggtgacca agggcggccc cctgcccttc acctgggaca tcctgagccc tcagttcatg
      841 tacggctcca atgcctacgt gaagcacccc gccgacatcc ccgactactt taagctgtcc
      901 ttccccgagg gcttcaagtg ggagcgcgtg atgaaattcg aggacggcgg cgtggtgacc
      961 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1021 accaacttcc cctccgacgg ccccgtgatg cagaagaaga ccatgggctg ggaggccctc
     1081 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg aggtcaagcc cagagtgaag
     1141 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     1201 gtgcagctgc ccggcgccta caacgtcaac cgcaagctgg acatcaccag ccacaacgag
     1261 gactacacca tcgtggagca gtacgagaga gccgagggcc gccactccac cggcggcatg
     1321 gacgagctgt acaagtccgg agaagggcaa gggcaagggc aagggccggg ccgcggctac
     1381 gcgtatcgat ccatgttcga ggcgcgcctg gtccagggct ccatcctcaa gaaggtgttg
     1441 gaggcactca aggacctcat caacgaggcc tgctgggata ttagctccag cggtgtaaac
     1501 ctgcagagca tggactcgtc ccacgtctct ttggtgcagc tcaccctgcg gtctgagggc
     1561 ttcgacacct accgctgcga ccgcaacctg gccatgggcg tgaacctcac cagtatgtcc
     1621 aaaatactaa aatgcgccgg caatgaagat atcattacac taagggccga agataacgcg
     1681 gataccttgg cgctagtatt tgaagcacca aaccaggaga aagtttcaga ctatgaaatg
     1741 aagttgatgg atttagatgt tgaacaactt ggaattccag aacaggagta cagctgtgta
     1801 gtaaagatgc cttctggtga atttgcacgt atatgccgag atctcagcca tattggagat
     1861 gctgttgtaa tttcctgtgc aaaagacgga gtgaaatttt ctgcaagtgg agaacttgga
     1921 aatggaaaca ttaaattgtc acagacaagt aatgtcgata aagaggagga agctgttacc
     1981 atagagatga atgaaccagt tcaactaact tttgcactga ggtacctgaa cttctttaca
     2041 aaagccactc cactctcttc aacggtgaca ctcagtatgt ctgcagatgt accccttgtt
     2101 gtagagtata aaattgcgga tatgggacac ttaaaatact acttggctcc caagatcgag
     2161 gatgaagaag gatcttaggg atccaccgga tctagataac tgatcataat cagccatacc
     2221 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2281 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2341 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2401 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     2461 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2521 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2581 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2641 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2701 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2761 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2821 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2881 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2941 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3001 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3061 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3121 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3181 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3241 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3301 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3361 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3421 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3481 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3541 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3601 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3661 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3721 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3781 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3841 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3901 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     3961 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4021 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4081 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4141 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4201 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4261 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4321 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4381 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4441 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4501 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4561 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4621 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4681 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4741 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4801 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4861 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4921 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4981 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5041 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5101 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5161 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5221 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5281 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5341 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5401 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5461 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     5521 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
