LOCUS       sequence-164254-        4695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: C1 Cloning Vector, Excitation: 505 / 569, Emission: 
            516 / 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4695)
  AUTHORS   Paez-Segala MG, Sun MG, Shtengel G, Viswanathan S, Baird MA, Macklin
            JJ, Patel R, Allen JR, Howe ES, Piszczek G, Hess HF, Davidson MW, 
            Wang Y, Looger LL
  TITLE     Fixation-resistant photoactivatable fluorescent proteins for CLEM.
  JOURNAL   Nat Methods. 2015 Jan 12. doi: 10.1038/nmeth.3225.
  PUBMED    25581799
REFERENCE   2  (bases 1 to 4695)
  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"; journalName: "Nat 
            Methods. 2015 Jan 12. doi: 10.1038/nmeth.3225."
COMMENT     Sequence Label: mEos4b-C1
FEATURES             Location/Qualifiers
     source          1..4695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1300
                     /codon_start=1
                     /product="fixation-resistant truly monomeric green-to-red 
                     photoswitchable fluorescent protein (Paez-Segala et al., 
                     2015)"
                     /label=mEos4b
                     /note="derived from mEos2"
                     /translation="MVSAIKPDMRIKLRMEGNVNGHHFVIDGDGTGKPYEGKQTMDLEV
                     KEGGPLPFAFDILTTAFHYGNRVFVKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNA
                     RNDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALL
                     LEGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHAIEILSHDKDYNKVKLYEHAVAHSGLP
                     DNARR"
     misc_feature    1301..1366
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1490..1611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1527..1546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1581..1600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1618..2073)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1755..1776)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1967..1986
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2100..2204
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2206..2563
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2206..2397
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2229..2249)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2414..2549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2598..3392
                     /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             2598..3392
                     /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(2652..2671)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3262..3281
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3580..3599)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3624..3671
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4000..4588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4489..4508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagtgc gattaagcca gacatgagga tcaaactccg
      661 tatggaaggc aacgtaaacg ggcaccactt tgtgatcgac ggagatggta caggcaagcc
      721 ttatgaggga aaacagacca tggatcttga agtcaaagag ggcggacctc tgccttttgc
      781 ctttgatatc ctgaccactg cattccatta cggcaacagg gtattcgtga aatatccaga
      841 caacatacaa gactatttta agcagtcgtt tcctaagggg tattcgtggg aacgaagctt
      901 gactttcgaa gacgggggca tttgcaatgc cagaaacgac ataacaatgg aaggggacac
      961 tttctataat aaagttcgat tttatggtac caactttccc gccaatggtc cagttatgca
     1021 gaagaagacg ctgaaatggg agccctccac tgagaaaatg tatgtgcgtg atggagtgct
     1081 gacgggtgat attgagatgg ctttgttgct tgaaggaaat gcccattacc gatgtgactt
     1141 cagaactact tacaaagcta aggagaaggg tgtcaagtta ccaggcgccc actttgtgga
     1201 ccacgccatt gagattttaa gccatgacaa agattacaac aaggttaagc tgtatgagca
     1261 tgctgttgct cattctggat tgcctgacaa tgccagacga tccggactca gatctcgagc
     1321 tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata
     1381 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     1441 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1501 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1561 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     1621 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1681 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     1741 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     1801 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     1861 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     1921 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     1981 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2041 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     2101 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2161 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     2221 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2281 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2341 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2401 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2461 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2521 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     2581 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2641 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2701 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2761 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2821 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2881 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2941 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3001 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3061 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3121 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3181 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3241 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3301 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3361 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3421 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3481 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3541 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     3601 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     3661 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3721 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3781 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     3841 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3901 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3961 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4021 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4081 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4141 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4201 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4261 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4321 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4381 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4441 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4501 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4561 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4621 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4681 taaccgtatt accgc
//
