LOCUS       sequence-329828-        6038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mRuby4(1-10) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6038)
  AUTHORS   Tamura R, Jiang F, Xie J, Kamiyama D
  TITLE     Multiplexed labeling of cellular proteins with split fluorescent 
            protein tags.
  JOURNAL   Commun Biol. 2021 Feb 26;4(1):257. doi: 10.1038/s42003-021-01780-4.
  PUBMED    33637968
REFERENCE   2  (bases 1 to 6038)
  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: "Commun 
            Biol."; date: "2021-02-26"; volume: "4(1):257. doi"; pages: " 
            10.1038/s42003-021-01780-4"
COMMENT     Sequence Label: pcDNA3.1-mRuby4(1-10)
FEATURES             Location/Qualifiers
     source          1..6038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1395..1412)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1401..1625
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1671..2099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1758..1777)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1968..1989
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2108..2128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2113..2442
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2293..2428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2355..2374
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2509..3303
                     /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(2563..2582)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3173..3192
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3477..3598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3514..3533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3568..3587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3647..3663)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3647..3663)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3660..3682)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3671..3687
                     /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(3695..3701,3702..3719,3720..3725))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3740..3761
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3878..3895)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4049..4637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4129..4148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4808..5599,5600..5668))
                     /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     5431..5450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5669..5773)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5848..5867)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccat ggtgtctaag ggcgaagagc tgatcaagga
      721 aaatatgcgt acgaaggtgg tcatggaagg ttcggtcaac ggccaccatt tcaaatgcat
      781 aggtgaagga gaaggcagac cgtacgaggg agtgcaaacc atgaggatca aagtcatcga
      841 gggaggaccc ctgccatttg cctttgacat tcttgccacg tcgttcatgt atggcagccg
      901 tacctttatc aagtacccgg ccgacatccc tgatttcttt aaacagtcct ttcctgaggg
      961 ttttacttgg gaaagagtta cgagatacga agatggtgga gtcatcaccg tcacgcagga
     1021 cacctgcctt gaggatggcg agctcgtcta caacgtcaag gtcagcgggg taaactttcc
     1081 ctccaatggt cccgtgatgc agaagaagac caagggttgg gagcctaata cagagatgat
     1141 gtatccagta gatggtggtc tgagaggata cactgacatc gcactgaaag ttgatggtgg
     1201 tggccatctg cactgcaact tcgtgacaac ttacaggtca aaaaagaccg tcgggaacat
     1261 caagatgccc ggtgtccatg ccgttgatca ccgcctggaa aggatcgagg agagtgacaa
     1321 ttaagaattc tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta
     1381 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     1441 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     1501 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1561 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     1621 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     1681 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1741 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1801 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1861 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1921 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1981 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2041 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     2101 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2161 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2221 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2281 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2341 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2401 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttat
     2461 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2521 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2581 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2641 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     2701 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     2761 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     2821 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     2881 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     2941 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3001 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3061 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3121 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3181 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3241 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3301 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3361 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3421 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3481 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3541 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3601 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3661 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3721 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3781 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3841 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3901 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3961 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4021 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4081 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4141 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4201 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4261 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4321 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4381 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4441 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4501 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4561 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4621 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4681 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4741 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4801 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4861 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4921 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4981 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5041 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5101 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5161 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5221 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5281 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5341 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5401 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5461 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5521 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5581 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5641 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5701 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5761 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     5821 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5881 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     5941 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     6001 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
