LOCUS       sequence-474129-        6144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of rsZACRO, a positively reversibly 
            photoswitchable fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6144)
  AUTHORS   Ozaki-Noma R, Wazawa T, Kakizuka T, Shidara H, Takemoto K, Nagai T
  TITLE     Positive-Type Reversibly Photoswitching Red Fluorescent Protein for 
            Dual-Color Superresolution Imaging with Single Light Exposure for 
            Off-Switching.
  JOURNAL   ACS Nano. 2025 Feb 25;19(7):7188-7201. doi: 10.1021/acsnano.4c16847.
            Epub 2025 Feb 12.
  PUBMED    39937184
REFERENCE   2  (bases 1 to 6144)
  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"; doi: 
            "10.1021/acsnano.4c16847"; journalName: "ACS Nano"; date: 
            "2025-02-25- 25"; volume: "19"; issue: "7"; pages: "7188-7201"
COMMENT     Sequence Label: rsZACRO/pcDNA3
FEATURES             Location/Qualifiers
     source          1..6144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1403..1412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1697..1715)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1698..1715)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1735..1752)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1741..1965
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2011..2439
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2098..2117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2308..2329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2448..2468)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2453..2782
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2633..2768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2695..2714
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2849..3643
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2903..2922)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3513..3532
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3817..3938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3854..3873)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3908..3927
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3987..4003)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3987..4003)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4000..4022)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4011..4027
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4035..4041,4042..4059,4060..4065))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4080..4101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4218..4235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4389..4977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4469..4488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5148..5939,5940..6008))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5771..5790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6009..6113)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cccaccatgg gttcggaggt cggccccgcc ctgttccagc gggacatgac
      961 cttcaaaatc ttcatcgacg gcgaggttaa cggccagaag ttcaccatcg tggccgacgg
     1021 cagcagcaag ttcccccacg gcgacttcaa cgtgcacgcc gtgtgcgaga ccggcaagct
     1081 gcccatgagc tggaagccca tctgccacct gatccagtac ggcgagccct tcttcgcccg
     1141 ctaccccgac ggcatcatcc atttcagtca ggagtgcttc cccgagggcc tgagcatcga
     1201 ccgcaccgtg cgcttcgaga acgacggcac catgaccagc caccacacct acgagctgga
     1261 cggcacctgc gtggtgagcc gcatcaccgt gaactgcgac ggcttcctgc ccgacggccc
     1321 catcatgcgc gaccagctgg tggacattcg tccctgcgag acccgcatac tcccctacgg
     1381 ccccaacgcc gtgcgccaga ccgccaccat gggcttcacc accgccgacg gcggcaaggt
     1441 gatgggccac ttcgacagca agatgacctt caacggcagc cgcgccatcg agatccccgg
     1501 cccacacttc gtgaccatca tcaccgagca gacgagggac cctagcgaca agcgcgacca
     1561 cgtgtgccag cgcgaggtgg cctacgccca cagcgggccc cgcaacacca gcgccatcgg
     1621 tagcgacgag gattaagaat tctgcagata tccatcacac tggcggccgc tcgagcatgc
     1681 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     1741 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     1801 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     1861 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     1921 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     1981 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2161 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2341 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2401 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     2461 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2521 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2581 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2641 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2701 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     2761 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     2821 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     2881 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2941 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3001 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3061 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3121 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3181 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3241 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3301 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3361 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3421 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3481 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3541 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3601 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3661 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3721 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3781 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     3841 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3901 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     3961 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4021 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4081 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4141 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4201 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4261 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4321 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4381 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4441 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4501 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4561 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4621 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4681 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4741 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4801 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4861 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4921 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4981 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5041 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5101 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5221 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5281 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5341 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5401 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5461 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5521 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5581 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5641 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5701 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5761 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5821 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5881 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5941 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6001 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6121 ccccgaaaag tgccacctga cgtc
//
