LOCUS       sequence-228942-        5824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus, Excitation: 505 / 569, Emission: 516 / 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5824)
  TITLE     Davidson Photoactivatable Fluorescent Proteins
REFERENCE   2  (bases 1 to 5824)
  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: tdEos-CENPA-N-22
FEATURES             Location/Qualifiers
     source          1..5824
                     /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"
     misc_feature    961..1013
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1027..1704
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     CDS             1753..2430
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="SAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVKE
                     GGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIARN
                     DITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIRMALLLE
                     GNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPDN
                     ARR"
     polyA_signal    2552..2673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2589..2608)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2643..2662
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2680..3135)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2817..2838)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3029..3048
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3162..3266
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3268..3625
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3268..3459
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3291..3311)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3476..3611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3660..4454
                     /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             3660..4454
                     /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(3714..3733)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4324..4343
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4642..4661)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4686..4733
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5062..5650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5551..5570
                     /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 atgggcccgc gccgccggag ccgaaagccc gaggccccga ggaggcgcag cccgagcccg
      601 accccgaccc ccggcccctc ccggcggggc ccctccttag gcgcttcctc ccatcaacac
      661 agtcggcgga gacaaggttg gctaaaggag atccgaaagc ttcagaagag cacacacctc
      721 ttgataagga agctgccctt cagccgcctg gcaagagaaa tatgtgttaa attcactcgt
      781 ggtgtggact tcaattggca agcccaggcc ctattggccc tacaagaggc agcagaagca
      841 tttctagttc atctctttga ggacgcctat ctcctcacct tacatgcagg ccgagttact
      901 ctcttcccaa aggatgtgca actggcccgg aggatccggg gccttgagga gggactcggc
      961 ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggtc
     1021 gccaccatga gtgcgattaa gccagacatg aagatcaacc tccgtatgga aggcaacgta
     1081 aacgggcacc actttgtgat cgacggagat ggtacaggca agccttttga gggaaaacag
     1141 agtatggatc ttgaagtcaa agagggcgga cctctgcctt ttgcctttga tatcctgacc
     1201 actgcattcc attacggcaa cagggtattc gccgaatatc cagaccacat acaagactat
     1261 tttaagcagt cgtttcctaa ggggtattcg tgggaacgaa gcttgacttt cgaagacggg
     1321 ggcatttgca ttgccagaaa cgacataaca atggaagggg acactttcta taataaagtt
     1381 cgatttcacg gtgtaaactt tcccgccaat ggtccagtta tgcagaagaa gacgctgaaa
     1441 tgggagccct ccactgagaa aatgtatgtg cgtgatggag tgctgactgg tgatattcgc
     1501 atggctttgt tgcttgaagg aaatgcccat taccgatgtg acttcagaac tacttacaaa
     1561 gctaaggaga agggtgtcaa gttaccaggc taccactttg tggaccactg cattgagatt
     1621 ttaagccatg acaaagatta caacaaggtt aagctgtatg agcatgctgt tgctcattct
     1681 ggattgcctg acaatgccag acgatccggt agcggtcacg gtactggttc tactggttct
     1741 ggttcttctg agagtgcgat taagccagac atgaagatca acctccgtat ggaaggcaac
     1801 gtaaacgggc accactttgt gatcgacgga gatggtacag gcaagccttt tgagggaaaa
     1861 cagagtatgg atcttgaagt caaagagggc ggacctctgc cttttgcctt tgatatcctg
     1921 accactgcat tccattacgg caacagggta ttcgccgaat atccagacca catacaagac
     1981 tattttaagc agtcgtttcc taaggggtat tcgtgggaac gaagcttgac tttcgaagac
     2041 gggggcattt gcattgccag aaacgacata acaatggaag gggacacttt ctataataaa
     2101 gttcgatttc acggtgtaaa ctttcccgcc aatggtccag ttatgcagaa gaagacgctg
     2161 aaatgggagc cctccactga gaaaatgtat gtgcgtgatg gagtgctgac tggtgatatt
     2221 cgcatggctt tgttgcttga aggaaatgcc cattaccgat gtgacttcag aactacttac
     2281 aaagctaagg agaagggtgt caagttacca ggctaccact ttgtggacca ctgcattgag
     2341 attttaagcc atgacaaaga ttacaacaag gttaagctgt atgagcatgc tgttgctcat
     2401 tctggattgc ctgacaatgc cagacgataa gcggccgcga ctctagatca taatcagcca
     2461 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2521 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2581 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2641 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     2701 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     2761 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     2821 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     2881 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     2941 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3001 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3061 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3121 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3181 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3241 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3301 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3361 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3421 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3481 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3541 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3601 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     3661 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3721 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3781 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3841 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3901 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3961 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4021 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4081 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4141 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4201 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4261 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4321 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4381 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4441 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4501 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4561 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4621 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4681 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     4741 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     4801 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     4861 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     4921 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4981 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5041 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5101 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5161 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5221 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5281 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5341 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5401 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5461 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5521 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5581 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5641 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5701 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     5761 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     5821 tccg
//
