LOCUS       sequence-489669-        5805 bp    DNA     circular SYN 17-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5805)
  AUTHORS   Subach FV, Patterson GH, Manley S, Gillette JM, Lippincott-Schwartz 
            J, Verkhusha VV
  TITLE     Photoactivatable mCherry for high-resolution two-color fluorescence 
            microscopy.
  JOURNAL   Nat Methods. 2009 Feb;6(2):153-9. Epub 2009 Jan 25.
  PUBMED    19169259
REFERENCE   2  (bases 1 to 5805)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods."; date: "2009-02"; volume: "6(2)"; pages: "153-9. Epub 2009
            Jan 25"
COMMENT     Sequence Label: pPAmCherry-b-actin
FEATURES             Location/Qualifiers
     source          1..5805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             607..1314
                     /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(752..770)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2593..2714
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2630..2649)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2684..2703
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2721..3176)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2858..2879)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3070..3089
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3203..3307
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3309..3666
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3309..3500
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3332..3352)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3517..3652
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3701..4495
                     /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             3701..4495
                     /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(3755..3774)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4365..4384
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4683..4702)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4727..4774
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5103..5691
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5592..5611
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcggtc
      601 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcattaagga gttcatgcgc
      661 ttcaaggtgc acatggaggg gtccgtgaac ggccacgtgt tcgagatcga gggcgagggc
      721 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
      781 ctgcccttca cctgggacat cctgagccct cagttcatgt acggctccaa tgcctacgtg
      841 aagcaccccg ccgacatccc cgactacttt aagctgtcct tccccgaggg cttcaagtgg
      901 gagcgcgtga tgaaattcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      961 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1021 cccgtgatgc agaagaagac catgggctgg gaggccctct ccgagcggat gtaccccgag
     1081 gacggcgccc tgaagggcga ggtcaagccc agagtgaagc tgaaggacgg cggccactac
     1141 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1201 aacgtcaacc gcaagctgga catcaccagc cacaacgagg actacaccat cgtggagcag
     1261 tacgagagag ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtccgga
     1321 ctcagatctc gagctatgga tgatgatatc gccgcgctcg tcgtcgacaa cggctccggc
     1381 atgtgcaagg ccggcttcgc gggcgacgat gccccccggg ccgtcttccc ctccatcgtg
     1441 gggcgcccca ggcaccaggg cgtgatggtg ggcatgggtc agaaggattc ctatgtgggc
     1501 gacgaggccc agagcaagag aggcatcctc accctgaagt accccatcga gcacggcatc
     1561 gtcaccaact gggacgacat ggagaaaatc tggcaccaca ccttctacaa tgagctgcgt
     1621 gtggctcccg aggagcaccc cgtgctgctg accgaggccc ccctgaaccc caaggccaac
     1681 cgcgagaaga tgacccagat catgtttgag accttcaaca ccccagccat gtacgttgct
     1741 atccaggctg tgctatccct gtacgcctct ggccgtacca ctggcatcgt gatggactcc
     1801 ggtgacgggg tcacccacac tgtgcccatc tacgaggggt atgccctccc ccatgccatc
     1861 ctgcgtctgg acctggctgg ccgggacctg actgactacc tcatgaagat cctcaccgag
     1921 cgcggctaca gcttcaccac cacggccgag cgggaaatcg tgcgtgacat taaggagaag
     1981 ctgtgctacg tcgccctgga cttcgagcaa gagatggcca cggctgcttc cagctcctcc
     2041 ctggagaaga gctacgagct gcctgacggc caggtcatca ccattggcaa tgagcggttc
     2101 cgctgccctg aggcactctt ccagccttcc ttcctgggca tggagtcctg tggcatccac
     2161 gaaactacct tcaactccat catgaagtgt gacgtggaca tccgcaaaga cctgtacgcc
     2221 aacacagtgc tgtctggcgg caccaccatg taccctggca ttgccgacag gatgcagaag
     2281 gagatcactg ccctggcacc cagcacaatg aagatcaaga tcattgctcc tcctgagcgc
     2341 aagtactccg tgtggatcgg cggctccatc ctggcctcgc tgtccacctt ccagcagatg
     2401 tggatcagca agcaggagta tgacgagtcc ggcccctcca tcgtccaccg caaatgcttc
     2461 tagggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     2521 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2581 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2641 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2701 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2761 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2821 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2881 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2941 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3001 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3061 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3121 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3181 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3241 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3301 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3361 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3421 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3481 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3541 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3601 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3661 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3721 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3781 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3841 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3901 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3961 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4021 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4081 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4141 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4201 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4261 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4321 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4381 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4441 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4501 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4561 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4621 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4681 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4741 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4801 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4861 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4921 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4981 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5041 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5101 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5161 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5221 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5281 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5341 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5401 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5461 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5521 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5581 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5641 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5701 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5761 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
