LOCUS       sequence-175700-        5604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus, Excitation: 555, Emission: 584.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5604)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 5604)
  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: mTagRFP-T2-PCNA-19
FEATURES             Location/Qualifiers
     source          1..5604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..27
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             31..51
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             55..75
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      82..91
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             88..819
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTETLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     polyA_signal    1792..1913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1829..1848)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1883..1902
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1920..2375)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2057..2078)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2269..2288
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2402..2506
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2508..2865
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2508..2699
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2531..2551)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2716..2851
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2900..3694
                     /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             2900..3694
                     /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(2954..2973)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3564..3583
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3882..3901)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3926..3973
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4302..4890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4791..4810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5065..5368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5369..5572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5523..5543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5599..5604,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt agatccaaaa
       61 aagaagagaa aggtaccggt cgccaccatg gtgtctaagg gcgaagagct gattaaggag
      121 aacatgcaca tgaagctgta catggagggc accgtgaaca accaccactt caagtgcaca
      181 tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag
      241 ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta cggcagcaga
      301 accttcatca accacaccca gggcatcccc gatttcttta agcagtcctt ccctgagggc
      361 ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac
      421 accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttccca
      481 tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccaacac cgagaccctg
      541 taccccgctg acggcggcct ggaaggcaga accgacatgg ccctgaagct cgtgggcggg
      601 ggccacctga tctgcaactt caagaccaca tacagatcca agaaacccgc taagaacctc
      661 aagatgcccg gcgtctacta tgtggaccac agactggaaa gaatcaagga ggccgacaaa
      721 gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa
      781 ctggggcaca aacttaatgg catggacgag ctgtacaagt ccggagaagg gcaagggcaa
      841 gggcaagggc cgggccgcgg ctacgcgtat cgatccatgt tcgaggcgcg cctggtccag
      901 ggctccatcc tcaagaaggt gttggaggca ctcaaggacc tcatcaacga ggcctgctgg
      961 gatattagct ccagcggtgt aaacctgcag agcatggact cgtcccacgt ctctttggtg
     1021 cagctcaccc tgcggtctga gggcttcgac acctaccgct gcgaccgcaa cctggccatg
     1081 ggcgtgaacc tcaccagtat gtccaaaata ctaaaatgcg ccggcaatga agatatcatt
     1141 acactaaggg ccgaagataa cgcggatacc ttggcgctag tatttgaagc accaaaccag
     1201 gagaaagttt cagactatga aatgaagttg atggatttag atgttgaaca acttggaatt
     1261 ccagaacagg agtacagctg tgtagtaaag atgccttctg gtgaatttgc acgtatatgc
     1321 cgagatctca gccatattgg agatgctgtt gtaatttcct gtgcaaaaga cggagtgaaa
     1381 ttttctgcaa gtggagaact tggaaatgga aacattaaat tgtcacagac aagtaatgtc
     1441 gataaagagg aggaagctgt taccatagag atgaatgaac cagttcaact aacttttgca
     1501 ctgaggtacc tgaacttctt tacaaaagcc actccactct cttcaacggt gacactcagt
     1561 atgtctgcag atgtacccct tgttgtagag tataaaattg cggatatggg acacttaaaa
     1621 tactacttgg ctcccaagat cgaggatgaa gaaggatctt agggatccac cggatctaga
     1681 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1741 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     1801 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1861 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     1921 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     1981 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2041 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2101 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2161 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2221 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2281 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2341 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     2401 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2461 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     2521 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2581 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2641 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2701 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2761 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2821 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     2881 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     2941 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3001 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3061 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3121 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3181 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3241 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3301 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3361 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3421 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3481 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3541 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3601 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3661 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3721 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3781 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3841 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     3901 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     3961 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4021 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4081 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4141 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4201 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4261 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4321 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4381 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4441 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4501 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4561 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4621 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4681 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4741 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4801 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4861 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4921 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4981 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     5041 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     5101 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     5161 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     5221 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     5281 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     5341 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     5401 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     5461 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     5521 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     5581 gaaccgtcag atccgctagc cacc
//
