LOCUS       sequence-173170-        5136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus, 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 5136)
  TITLE     Michael Davidson Unpublished
REFERENCE   2  (bases 1 to 5136)
  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: mEos4b-CenpA-C-22
FEATURES             Location/Qualifiers
     source          1..5136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..681
                     /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"
     polyA_signal    1312..1433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1349..1368)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1403..1422
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1440..1895)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1577..1598)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1789..1808
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1922..2026
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2028..2385
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2028..2219
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2051..2071)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2236..2371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2420..3214
                     /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             2420..3214
                     /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(2474..2493)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3084..3103
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3402..3421)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3446..3493
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3822..4410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4311..4330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4585..4888
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4889..5092
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5043..5063
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(5131..5136,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagtg cgattaagcc agacatgagg atcaaactcc gtatggaagg caacgtaaac
       61 gggcaccact ttgtgatcga cggagatggt acaggcaagc cttatgaggg aaaacagacc
      121 atggatcttg aagtcaaaga gggcggacct ctgccttttg cctttgatat cctgaccact
      181 gcattccatt acggcaacag ggtattcgtg aaatatccag acaacataca agactatttt
      241 aagcagtcgt ttcctaaggg gtattcgtgg gaacgaagct tgactttcga agacgggggc
      301 atttgcaatg ccagaaacga cataacaatg gaaggggaca ctttctataa taaagttcga
      361 ttttatggta ccaactttcc cgccaatggt ccagttatgc agaagaagac gctgaaatgg
      421 gagccctcca ctgagaaaat gtatgtgcgt gatggagtgc tgacgggtga tattgagatg
      481 gctttgttgc ttgaaggaaa tgcccattac cgatgtgact tcagaactac ttacaaagct
      541 aaggagaagg gtgtcaagtt accaggcgcc cactttgtgg accacgccat tgagatttta
      601 agccatgaca aagattacaa caaggttaag ctgtatgagc atgctgttgc tcattctgga
      661 ttgcctgaca atgccagacg atccggactc agatctcgag ctcaagcttc gaattctagc
      721 agttctagag taagcagcag agtaagtggc ccgcgccgcc ggagccgaaa gcccgaggcc
      781 ccgaggaggc gcagcccgag cccgaccccg acccccggcc cctcccggcg gggcccctcc
      841 ttaggcgctt cctcccatca acacagtcgg cggagacaag gttggctaaa ggagatccga
      901 aagcttcaga agagcacaca cctcttgata aggaagctgc ccttcagccg cctggcaaga
      961 gaaatatgtg ttaaattcac tcgtggtgtg gacttcaatt ggcaagccca ggccctattg
     1021 gccctacaag aggcagcaga agcatttcta gttcatctct ttgaggacgc ctatctcctc
     1081 accttacatg caggccgagt tactctcttc ccaaaggatg tgcaactggc ccggaggatc
     1141 cggggccttg aggagggact aggctaaggt accgcgggcc cgggatccac cggatctaga
     1201 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1261 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     1321 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1381 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     1441 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     1501 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     1561 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     1621 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     1681 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     1741 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     1801 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     1861 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     1921 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1981 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     2041 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2101 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2161 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2221 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2281 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2341 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     2401 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     2461 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     2521 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     2581 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     2641 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     2701 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     2761 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     2821 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     2881 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     2941 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3001 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3061 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3121 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3181 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3241 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3301 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3361 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     3421 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     3481 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     3541 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3601 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     3661 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3721 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3781 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3841 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3901 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3961 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4021 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4081 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4141 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4201 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4261 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4321 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4381 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4441 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4501 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     4561 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     4621 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     4681 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     4741 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     4801 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     4861 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     4921 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     4981 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     5041 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     5101 gaaccgtcag atccgctagc gctaccggtc gccacc
//
